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WO2017074159A2 - Coumestrol-containing germinated fermented soybean extract having hepatocyte protective effect and bone density increase effect and composition containing same - Google Patents

Coumestrol-containing germinated fermented soybean extract having hepatocyte protective effect and bone density increase effect and composition containing same Download PDF

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Publication number
WO2017074159A2
WO2017074159A2 PCT/KR2016/012399 KR2016012399W WO2017074159A2 WO 2017074159 A2 WO2017074159 A2 WO 2017074159A2 KR 2016012399 W KR2016012399 W KR 2016012399W WO 2017074159 A2 WO2017074159 A2 WO 2017074159A2
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fermented soybean
germinated fermented
soybean extract
germinated
extract
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WO2017074159A3 (en
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์ด๋™์„ฑ
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Hubio
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Hubio
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methanthelineย 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/12—Drugs for genital or sexual disorders; Contraceptives for climacteric disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment

Definitions

  • the present invention relates to cousterol-containing germinated fermented soybean extract having a hepatocellular protective effect and an effect of increasing bone density and a composition comprising the same. More specifically, the in vitro antioxidant activity, cell viability, hematological test of animal models, MDA (malone) for the hepatotoxicity of menopausal disorders using germinated fermented soybean extract as a component that improves lipids and menopausal disorders in natural foods Dialdehyde), GST (glutathione S-transferase), GSH (reduced glutathione), SOD (Superoxide dismutase), and catalase were measured to verify hepatocellular protective effects.
  • MDA malone
  • the germinated fermented soybean extract as a sample in vivo
  • Fermented germinated soybean extract of the present invention significantly increases blood estrogen and testosterone concentrations, thereby relieving menopausal symptoms, and increasing bone mineral protection and increasing bone mineral content in vitro and in vivo. Increase, and the thickness and number of spongy bones increase
  • Didzein in legumes isoflavones contain cumesterol, which regulates female hormone balance and content to prevent breast and uterine cancer, slowing menopause and alleviating many of the symptoms associated with it.
  • the aging of the population is progressing very rapidly all over the world, and especially in Korea, the progress of aging is very fast compared to other developed countries.
  • women's average life expectancy is increased and the intellectual level is improved as a result of the health promotion resulting from economic growth
  • interest in women's menopause and postmenopausal quality of life and health improvement is also increasing.
  • the main symptoms of menopause include hot flashes, anxiety, depression and nervousness. Symptoms such as cardiovascular disease, osteoporosis and memory loss are known. The cause of these symptoms is known to be caused by interrupted production and secretion of estrogen.
  • hormone replacement therapy has been proposed, but there are no results on usage, dose, and safety.
  • Phytoestrogen a natural food product, is a reproductive hormone-like substance that is beginning to attract attention for the prevention of menopausal symptoms and osteoporosis, and has been studied and reported to be able to substitute estrogen in postmenopausal women.
  • synthetic estrogen replacement therapy is currently used to treat menopausal symptoms, most women experience side effects such as remenstruation, breast softening and weight gain, and food intake with plant-based estrogen-like substances in natural foods. Relieving menopausal symptoms will help to alleviate the side effects.
  • Phytoestrogens include isoflavones and lignans and are compounds similar to those naturally occurring in plants such as beans, fruits, vegetables, peanuts and grains. Isoflavones are attracting attention as a kind of phytoestrogens that weakly bind to estrogen receptors in the body and have various physiological effects. Isoflavones extracted from soybeans exist mainly as glycosides in plants, and are absorbed into the human body to be converted into aglycone forms of genistein and dyedzein by glycosidase and have estrogenic properties.
  • composition for the prevention and treatment of menopausal symptoms comprising a soybean extract containing cumestrol as an active ingredient
  • a menopause containing a soybean extract containing cumestrol as an active ingredient It is the experimental result of the cell experiment (in vitro) on the composition for the prevention of diseases, and to improve the menopausal symptoms improvement and liver cell protection effect through the animal experiment (in-vivo) to confirm the cell experiment in the present invention. And improved germinated fermented soybean extract to achieve advanced methods and results.
  • Patent Document 1 Korean Patent Publication No. 10-2014-0053471
  • Non-Patent Document 1 Starzec, J, J, and D.F. Berger, Effects of stress and ovariectomy on the plasma cholesterol, serum triglyceride, and aortic cholesterol levels of female rats, Physiol Behav. 1986.37 (1): p. 99-104.
  • Non-Patent Document 2 Ettinger, B., Overview of estrogen replacement therapy a historical perspective. Proc Soc Exp Biol Med. 217 (1): p. 2-5.
  • Non-Patent Document 3 3. Baker, M.E., K.L. Medlock, and D.M. Sheehan, Flavonoids inhibit estrogen binding to rat alpha-fetoprotein. Proc Soc Exp Biol Med. 1998.217 (3): p.317-21.
  • Non-Patent Document 4 Boker, L.K., et al., Intake of Dietary Phytoestrogens by Dutch women. J Nutr. 2002. 132 (6): p. 1319-28.
  • Non-Patent Document 5 Morito, K., et al., Interaction of Phytoestrogens with estrogen receptors alpha and beta, Biol Pharm Bull. 2001. 24 (4); p.6-11.
  • Non-Patent Document 6 6. Albertazzi, P., et al., The effect of dietary soy supplementation on hot flushes, Obstet Gynecol. 1988. 91 (3): p. 389-94.
  • Non-Patent Document 7 7. Han, K, K., et al., Benefits of soy isoflavone therapeutic regimen on menopausal symptoms, Obstet Gynecol. 2002. 99 (3): p. 389-94.
  • Non-Patent Document 8 Petri Nahas, E., et al., Benefits of soy germ isoflavones in postmenopausal women with contraindication for conventional hormone replacement therapy, Maturitas. 2004. 48 (4): p.372-80.
  • Non-Patent Document 9 Kaari, C., ey al., Randomized clinical trial comparing conjugated equine estrogens and isoflavones in postmenopausal women: a pilot study, Maturitus. 2006. 53 (1): p. 49-58.
  • Non-Patent Document 10 Cheng, G., et al., Isoflavone treatment for acute menopausal symptoms, Menopause. 2007.14 (3Pt1): p.468-73.
  • Non-Patent Document 11 DJ., Et al., Effect of soy protein foods on low-density lipoprotein oxidation and ex vivo sex hormone receptor activity--a controlled crossover trial. Metabolism. 2000.49 (4): p.537-43.
  • Nonpatent Document 12 Kotsopoulos, D., et al., The effects of soy protein containing phytoesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.55 (21): p. 8589-95.
  • Non-Patent Document 13 Wangen, K.E., et al., Soy Isoflavones improve plasma lipids in normocholesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.73 (2): p.225-31.
  • Non-Patent Document 14 Feng, S., et al., Fungal-stressed germination of black soybeans leads to generation of oxooctadecadienoic acids in addition to glyceollins, J Agric Food Chem. 2007.55 (21): p.8589-95.
  • Non-Patent Document 15 15. Rosalki, S.B. and A.Y.Foo. Two new methods for seperating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma, Cli Chem. 1984. 30 (7): p.1182-6.
  • the present invention uses fermented germinated soybean extract to relieve menopausal symptoms, to protect the liver cells, and to increase the mineral content of bone.
  • menopause As women's average life expectancy is increased and the intellectual level is improved as a result of the health promotion resulting from economic growth, interest in women's menopause and postmenopausal quality of life and health improvement is also increasing.
  • the main symptoms of menopause include hot flashes, anxiety, depression and nervousness. Symptoms such as cardiovascular disease, osteoporosis and memory loss are known. The cause of these symptoms is known to be caused by interrupted production and secretion of estrogen.
  • hormone replacement therapy As a preventive measure, hormone replacement therapy has been proposed, but there are no results on usage, dose, and safety. In addition, there is a lack of appropriate drugs or health foods that have hepatoprotective effect and bone mineral density increase effect in postmenopausal women.
  • the present invention relates to a cousterol-containing germinated fermented soybean extract having an effect of protecting liver cells and increasing bone density in postmenopausal women and a composition comprising the same.
  • the germinated fermented soybean extract containing cumesterol to verify the effect of hepatocellular protection and bone density increase in postmenopausal women.
  • the germinated fermented soybean extract as a sample in vivo
  • blood calcium and phosphorus concentrations, ALP activity, bone mineral content, and bone mineral content were measured using Micro-CT.
  • the present invention was found to be effective for the protection of hepatocytes by measuring in vitro antioxidant activity, cell viability and hematological tests, MDA, GST, GSH, SOD, CAT, etc. for hepatotoxicity in menopausal disorders. It was also confirmed that the administration of germinated fermented soybeans increased the mineral content of bone. Bone density was significantly increased in the rats treated with germinated fermented soybean, and the thickness and number of spongy bones were increased. Significant increases in blood estrogen levels can be expected to replace the role of estrogens in germinated fermented soybeans, thereby alleviating menopausal symptoms.
  • the administration of germinated fermented soybean is expected to help improve menopausal obesity, and the high concentration of germinated fermented soybean is similar to that of estrogen. It was confirmed that there is a preventive effect of osteoporosis.
  • the oxidative damage of the body to oxidative stress which is the cause of various diseases in humans, requires antioxidant enzymes such as catalase, SOD, GSH, and GST.
  • antioxidant enzymes such as catalase, SOD, GSH, and GST.
  • As a result of measuring liver enzyme antioxidant activity using an animal model with artificial liver damage using germinated fermented soybean it was significantly reduced and increased compared to liver damage model administered only t- BHP such as SOD, GSH, GST, and MDA. Indicated.
  • the germinated fermented soybean was found to have a liver protective effect against oxidative stress.
  • Figure 1 shows the weight gain of the group receiving the estrogen.
  • Figure 2 shows serum lipids, cholesterol and glucose concentrations
  • Figure 4 shows the bone metabolic index measurement results.
  • Figure 5 shows the results of bone salt measurement.
  • Figure 7 shows a 2D image taken by using the Micro-CT.
  • Figure 8 shows the separation process using sep-pak of 70% ethanol germinated fermented soybeans.
  • Figure 9 shows the cell viability of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
  • Figure 10 shows the growth rate of t- BHP cells of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
  • Figure 11 shows the results of ROS measurement of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
  • Figure 12 shows the results of animal model hematology test AST (GOT), ALT (GPT).
  • Figure 13 shows the results of MDA measurement in Liver of the group administered t- BHP after the administration of germinated fermented soybean.
  • Figure 14 shows the results of GST measurement in Liver of the group administered t- BHP after administration of germinated fermented soybeans.
  • Figure 15 shows the results of GSH measurement in Liver of the group administered t- BHP after administration of germinated fermented soybean.
  • Figure 16 shows the results of SOD measurement in Liver of the group administered t- BHP after the administration of germinated fermented soybean.
  • Figure 17 shows the results of CAT measurements in Liver of the group administered t- BHP after the administration of germinated fermented soybeans.
  • Germinated fermented soybean used in this experiment was provided by Hubio. Soybeans obtained from Jeju Agricultural Research and Extension Services are germinated and fermented by inoculation with Aspergillus oryzae according to previous studies. After the extraction process using water washing, choice and ultrasonic, it was dried and used as a test sample.
  • the experimental animals were purchased from Korea Biolink (Daejeon), 8-week-old Sprague-Dawley female rats, and were bred as solid feed (Purina) in the laboratory. Therefore, the average body weight of each group was 289.72 โ‡ 0.86 g, and 7 animals of 6 groups (Negative control, Non-ovarian control group (sham), Estrogen group (OVX-estrogen), Germinated fermented soybean extract 0.5 g / kg Administered group (OVX-0.5), germinated fermented soybean extract 1.0 g / kg administered group (OVX-1.0) and germinated fermented soybean extract 2.0 g / kg administered group (OVX-2.0)].
  • 6 groups Negative control, Non-ovarian control group (sham), Estrogen group (OVX-estrogen), Germinated fermented soybean extract 0.5 g / kg Administered group (OVX-0.5), germinated fermented soybean extract 1.0 g / kg administered group (
  • the experimental animals were kept in a laboratory temperature of 22-24 ยฐ C, humidity 60 โ‡ 5% using a breeding cage (20 โ‡ 26 โ‡ 13 cm), and the day and night cycle (12 hours light / 12 hours dark) was controlled by an automatic control device. It was raised in the university animal room and approved by Korea University's Animal Experiment Ethics Committee, and managed according to the 'Guide for the care and use of Laboratory Animals' (NRC). Water and diet were freely supplied, and the experimental samples were dissolved in tertiary distilled water and orally administered 3 ml daily, and the control group (Negative-control, sham) was orally administered daily with the same dose of tertiary distilled water.
  • NRC Laboratory Animals'
  • ovarian resection was performed by dividing the group by randomized complete block design according to body weight. Surgery was performed with isoflurane and after anesthesia, the dorsal flank was incised to remove the ovary and the incision was closed. In the case of non-ovarian resection control (Sham), fat was removed from the ovary in the same way and the incision was closed. Samples were administered orally after having a recovery period of 2 weeks after surgery.
  • TG triglyceride
  • TCHO total cholesterol
  • HDL high density lipoprotein cholesterol
  • LDL low density lipoprotein cholesterol
  • GLU glucose
  • Plasma levels of estradiol and testosterone were measured by the rat estradiol Elisa kit (Enzo life science, NY, USA) and rat testosterone Elisa kit (Enzo life science, NY, USA) by ELISA.
  • Plasma calcium and phosphorus concentrations and alkaline alkaline phosphatase (ALP) concentrations were measured by FUJI DRI-CHEM 3500 and BONE-alkaline phosphatase (BONE-ALP) as bone markers.
  • the femur was dried and crushed through the fat removal process, and then the bone salt in the femur was measured.
  • a 2 mm high part of the tibia of the sacrificial rats was photographed using Micro-CT (Skyscan1072, SKYSCAN, Antwerpen, Belgium).
  • the imaging tube voltage was 50 kVp
  • the tube current was 500 โ‡ A
  • 0.5 mm aluminum filtration
  • the imaging time was 210 ms
  • the pixel size was 18 โ‡ m.
  • the threshold value was fixed at 58 to separate the bone column and bone marrow from the individual images, and the 3D image was reconstructed.
  • Bone microstructure indicators were analyzed using Skyscan TM CT-analyzersoftware from images taken with Micro-CT.
  • the indicators analyzed were Percent bone volume (BV / TV (%)), trabecular number (Tb.N ()), trabecular thickness (Tb.Th ( โ‡ m)), trabecular separation (Tb.Sp ( โ‡ m)), trabecular pattern factor (Tb.pf ()), and structure model index (SMI).
  • the data obtained were processed using the statistical package for social science (version 12.0) statistical program. T-test was performed on the negative-control group to identify the significant differences among the groups. The measured values are expressed as average standard error, and p โ‡ 0.05, p โ‡ 0.01, and p โ‡ 0.001 are indicated by *, **, and *** , respectively.
  • the weight gain of the estrogen-treated group was 12.1 โ‡ 8.7g, which tended to decrease relatively, but there was no significant difference in the germinated fermented soybean group. There was no significant difference in dietary intake changes.
  • the ovarian resection control group, non-ovarian resection control group, and the estrogen-administered group were labeled as Neg.C, Sham, and Estradiol, and the germinated fermented soybean 0.5g / kg, 1.0g / kg, and 2.0g / kg groups were 0.5, 1.0, and 2.0, respectively. Notation.
  • the figures are expressed as mean โ‡ standard error.
  • estrogen secretion decreases rapidly in the ovarian resection group. At this time, the body accumulates a lot of fat cells capable of producing estrogen in order to enhance the reduced estrogen secretion, which leads to menopausal obesity. Lipid, cholesterol, and glucose concentrations in blood were identified as indicators of menopausal obesity, and the results are shown in FIG. 2.
  • Neutral fat levels of the groups administered with 0.5 g / kg, 1.0 g / kg and 2.0 g / kg of germinated fermented soybean were 62.5 โ‡ 21.3 mg / dl, 59.0 โ‡ 8.5 mg / dl and 55.8 โ‡ 8.8 mg / dl, respectively.
  • the decrease was similar to that of the non-ovarian ablation control 66.8 โ‡ 12.6 mg / dl.
  • the blood glucose concentration was also 98.3 โ‡ 16.1 mg / dl, 123.3 โ‡ 14.4 mg / dl, 112.8 โ‡ 6.6 mg / dl in all groups administered with germinated fermented soybeans, the blood glucose concentration of 162.7 โ‡ It was confirmed that the tendency to decrease compared to 18.6 mg / dl.
  • the administration of germinated fermented soybean is expected to help improve menopausal obesity.
  • GOT and GPT are normal enzymes made in the liver. When the liver is damaged, these cells are broken down and the enzymes are released into the blood, increasing more than normal, so they are used as representative indicators of liver toxicity. Looking at Table 1 below, serum GOT, GPT activity analysis does not have a significant difference between groups. Therefore, it can be seen that the treatment of the sample does not show liver toxicity.
  • the cause of various menopausal symptoms is a decrease in estrogen production and secretion due to aging of the ovary. Therefore, it is important to confirm the increase of blood estrogen concentration in order to alleviate the menopausal symptoms of the administration of germinated fermented soybeans.
  • the estrogen concentration of the ovarian resection control group was 500.3 โ‡ 27.8 pg / ml compared to the estrogen concentration of 1020. โ‡ 63.6 pg / ml of the non-ovarian resection group, resulting in a significant (p โ‡ 0.001) decrease in estrogen concentration. You can see that. In the estrogen-administered group, it was confirmed that blood estrogen concentration was increased to 1134.1 โ‡ 205.8 pg / ml, which is similar to that of the non-ovarian ablation control group (FIG. 3).
  • Testosterone is a representative anabolic hormone that is involved in muscle and fat metabolism. Lower levels of these hormones reduce muscle mass in the body, which leads to lower basal metabolism and easier obesity. It also induces skin keratinization, one of the typical symptoms of menopause.
  • the concentration of testosterone in blood was decreased in all the groups that underwent ovarian ablation compared to the non-ovarian ablation control group. Among them, the testosterone concentrations of 2797.8 โ‡ 517.3 pg / ml and 1969.8 โ‡ 164.2 pg / ml were found in the estrogen-administered group and 1.0 g / kg of germinated fermented soybean, respectively. It was confirmed that the results were significantly increased (p โ‡ 0.05) compared to the control group.
  • Alkaline phosphatase is a biomarker of osteoblasts involved in bone metabolism. In other diseases, such as osteoporosis, in which bone metabolism increases rapidly, the level of ALP also increases.
  • the blood ALP level of the ovarian control group was 454.0 โ‡ 69.9 U / L, which was higher than that of the non-ovarian control group (340.2 โ‡ 33.31 U / L).
  • the group administered 0.5 g / kg of germinated fermented soybeans 260.0 โ‡ 646.9 U / L, significantly lower ALP activity than the control group (p โ‡ 0.05).
  • the Bone-ALP level was 140.8 โ‡ 58.5 U / L. In the group administered with 0.5 g / kg of germinated fermented soybean compared to the control group of ovarian resection, indicating that it was significantly lower (p โ‡ 0.05). .
  • the bone inflammation was significantly lower (p โ‡ 0.05) in the ovarian resection group compared with the non-ovarian resection control group.
  • the bone salts were 61.3 โ‡ 0.36% and 60.8 โ‡ 0.31%, respectively. This is a significant (p โ‡ 0.05) increase in bone salt compared to ovarian resection control. This confirmed that the germination of fermented soybeans increased the mineral content of bone.
  • the ovarian control group showed a low value of 1.17 โ‡ 0.09 mm -1 , but increased to 1.69 โ‡ 0.15 mm -1 similar to that of the estrogen group in the 2.0 g / kg germinated fermented soybean group. It could be confirmed (p โ‡ 0.05).
  • the ovarian control group showed a low value of 0.12 โ‡ 0.00 mm โ‡ 1 , but Tb.Th was significantly increased at all concentrations of estrogen and germinated fermented soybeans (FIG. 6). This means that the bone density of the germinated fermented soybeans increased significantly, and the thickness and number of spongy bones increased.
  • the MTT assay is based on the principle that the yellow soluble tetraz olium salt is reduced to the blue insoluble formazan product by mitochondrial succina dehydrogenase in living cells.
  • the absorbance of formazan is maximized at a wavelength of 540 nm.
  • the absorbance measured at this wavelength reflects the concentration of living and metabolically active cells and is used as an indicator of mitochondrial activity.
  • MTT is a test for measuring the mitochondrial activity to determine how toxic to a test substance by cytotoxicity test. If the test substance is toxic and the mitochondria are destroyed, the absorbance value is low.
  • ROS Reactive oxygen species
  • DCFH-DA Dichlorofluorescin diacetate
  • ROS free radicals
  • 70% EtoH germinated fermented soybean extract was incubated for 12 hours in a non-cytotoxic concentration and t- BHP 1 mM was treated for 1 hour to induce oxidative stress. . After 25 โ‡ M of DCFH-DA was added for 60 minutes, the fluorescence of DCF was measured.
  • FIG. 1 compared with Normal (100 โ‡ 1.00%) fluorescence increased by (267.62 โ‡ 2.68%) by t-BHP treatment, 0.5 (195.24 โ‡ 1.95%) by 70% EtoH germinated fermented soybean extract pretreatment , 0.25mg / mL (195.24 โ‡ 1.95), 0.125mg / mL (240 โ‡ 2.41), 0.0625mg / mL (252.80 โ‡ 2.53), significantly inhibited the concentration from the highest concentration to 70% EtoH germinated fermented soybean extract It was confirmed that the active oxygen reduction effect by.
  • LDH is a glycolytic enzyme and widely distributed in each tissue in the body. Especially, LDH in serum, liver, kidney, and muscle increases in heart disease, liver disease, malignancy, and leukemia, but has low specificity. In acute hepatitis, it escapes from hepatocytes like AST and ALT and increases in early acute hepatitis. In chronic hepatitis and cirrhosis, it increases to normal or mild. Significant differences were found in AST and ALT, one of the indicators of liver failure.
  • ALT and AST are hepatocellular leak enzymes, which are used for hepatotoxicity studies because enzymes are released into the blood as hepatic necrosis and hepatic tissue destruction progress. 12 shows the results of measuring ALT and AST activity.
  • the concentration of AST was 0.5 g / kg (83.20 โ‡ 2.86 U / L) and 1 g / kg (81.80 โ‡ 6.72 U / L), but there was no significant change in the concentration of germinated fermented soybeans. Significantly lower compared to the group (94.25 โ‡ 7.37 U / L).
  • the ALT showed no significant change in the concentration of germinated fermented soybean at 0.5 g / kg (33.71 โ‡ 5.59 U / L) and 1 g / kg (38.17 โ‡ 4.49 U / L), but only the t-BHP was treated in the sample treatment group. Significantly lower compared to (48.25 โ‡ 16.19 U / L).
  • Lipid peroxidation refers to a phenomenon in which polyunsaturated fatty acids in cell membranes are peroxidated by various free radicals derived from superoxide anion radicals that can occur naturally in a living body. Unsaturated fatty acids are degraded through peroxidation to produce MDA, which is an indicator of lipid peroxidation. 13, in the case of liver tissue, the group administered t- BHP after the administration of germinated fermented soybeans was significantly different between the groups at the sample concentration of 0.5 g / kg (34.25 โ‡ 5.80) and 1 g / kg (32.68 โ‡ 12.57). There was no difference, but it decreased as the concentration increased and decreased compared to the t- BHP-only group (44.66 โ‡ 1.94).
  • GST is an enzyme that exists in almost every cell of the body and is involved in many metabolism as well as detoxification of the liver. In particular, GST catalyzes the reaction of binding an electron-affinity substrate to GSH.
  • FIG. 14 GST measurements germination fermentation after administration of the beans treated group, t -BHP 0.5g / kg, 1g / kg, only one group administered t -BHP (25.68 โ‡ 4.28) Sample administration as compared to the concentration At 0.5 g / kg (33.65 โ‡ 12.97) and 1 g / kg (48.43 โ‡ 8.90), the sample concentration increased with increasing sample concentration, but significant difference with the t-BHP group was only observed at 1 g / kg.
  • GSH is a very important antidote and powerful antioxidant in the human body that plays a key role in detoxification, immune function, and antioxidant function in the body.
  • FIG. 13 GSH measurements germination fermentation after administration of the beans treated group, t -BHP 0.5g / kg, 1g / kg, t -BHP only administered group 21.07 โ‡ 8.33)
  • Sample administration concentration of 0.5 as compared to the There was no significant increase as the sample concentration increased at g / kg (25.12 โ‡ 2.44) and 1 g / kg (24.40 โ‡ 2.52).
  • there was no significant difference between the groups receiving the sample and the t- BHP group but there was a slight increase in the group receiving the sample.
  • SOD is an enzyme that reduces superoxide anion (O 2 ) to H 2 O 2 , and is present in cytosol and mitochondria. Only Cu / Zn-SOD exists in the cytosol, but Mn-SOD exists in the matrix of mitochondria, and a small amount of Cu / Zn-SOD exists between the inner and outer layers. SOD measurement results (Fig.
  • Catalase is an enzyme that protects the body from oxygen poison by reducing H 2 O 2 produced by the action of SOD or O 2 reduction to non-toxic H 2 O. Usually, it is present in peroxisome and in small amounts in mitochondria.
  • CAT measurement results (Fig. 10) after germination fermented beans treated with the group treated with t -BHP 0.5g / kg, 1g / kg, t -BHP only one dose group (182.24 โ‡ 19.29) administration sample concentration of 0.5 as compared to the At g / kg (204.49 โ‡ 41.51) and 1 g / kg (293.19 โ‡ 61.02), the sample concentration increased significantly.
  • the present invention may include one or more selected from the group consisting of excipients (disintegrants, binders, lubricants), stabilizers, emulsifiers, additives, etc., which are acceptable under the pharmacological, health functional food method, food hygiene method, in addition to the extract of germinated fermented soybean. Can be.
  • the composition may be made and used for oral use such as warnings, granules, powders, tablets, or capsules.
  • the dosage of the composition should be adjusted according to the criteria set forth in the relevant laws (use, purpose of use, patient's condition, age, sex, weight, prescription drug, disease).
  • the formulation was prepared by mixing the germinated fermented soybean extract with excipients or carriers acceptable in the pharmacological method to prepare powders, tablets, capsules, pills, and granules as preparations.
  • Germinated fermented soybean extract 100 mg, corn starch 100 mg, lactose 100 mg and stearic acid magnesium 2 mg were mixed and then compressed into tablets.
  • Germinated fermented soybean extract 100 mg, corn starch 100 mg, lactose 100 mg, magnesium stearate 2 mg was mixed, and then prepared by filling in gelatin capsules.
  • Foods were prepared by mixing the germinated fermented soybean extract with excipients or carriers allowed by the food hygiene or dietary supplement method.
  • Germinated fermented soybean extract 1g Vitamin A 70 โ‡ g, Vitamin E 1.0 mg, Vitamin 0.13 mg, Vitamin B2 0.15 mg, Vitamin B6 0.5 mg, Vitamin B12 0.2 โ‡ g, Vitamin C 10 mg, Biotin 10 โ‡ g, Nicotinamide 1.7 mg, Folic acid 50 โ‡ g, calcium pantothenate 0.5 mg, mineral mixture appropriate amount, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monophosphate 15 mg, calcium diphosphate 55 mg, potassium citrate 90 mg, carbonate Health food was prepared by mixing 100 mg of calcium and 24.8 mg of magnesium chloride.
  • the proportion of the vitamin and minerals mixed with the germinated fermented soybean extract may be variously modified according to the formulation or the consumer layer.
  • Germinated fermented soybean extract 1%, liquid fructose 0.5%, oligosaccharide 2%, sugar 2%, salt 0.5% and the balance of the purified water and subsidiary materials were combined and sterilized and then packaged to prepare a beverage.
  • the ratio of sugars, dietary fiber, vitamins and minerals to the germinated fermented soybean extract may be variously modified depending on the dosage form or consumer layer.
  • Germinated fermented soybean extract of the present invention significantly increases blood estrogen and testosterone concentrations, thereby alleviating menopausal symptoms, increasing bone mineral content, significantly increasing bone density, and the thickness and number of sponges Increases. Therefore, there is industrial applicability.

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Abstract

The present invention relates to a germinated fermented soybean extract containing coumestrol, a preparation method therefor, and a composition containing the germinated fermented soybean extract. It can be seen that the germinated fermented soybean extract of the present invention is effective in the hepatocyte protection against hepatotoxicity through in vitro antioxidative activity and cell viability, and hematological tests and MDA, GST, GSH, SOD, and CAT measurements in animal models. In addition, the germinated fermented soybean extract of the present invention increases the mineral content in bones, thereby significantly increasing the bone density, and has an effect of increasing the thickness and number of spongy bones. Furthermore, the germinated fermented soybean extract of the present invention has an effect of relieving symptoms of menopause.

Description

๊ฐ„์„ธํฌ ๋ณดํ˜ธ ํšจ๊ณผ ๋ฐ ๊ณจ ๋ฐ€๋„ ์ฆ๊ฐ€ ํšจ๊ณผ๋ฅผ ๊ฐ–๋Š” ์ฟ ๋ฉ”์Šคํ…Œ๋กค ํ•จ์œ  ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ ๋ฐ ์ด๋ฅผ ํฌํ•จํ•˜๋Š” ์กฐ์„ฑ๋ฌผCumesterol-containing germinated fermented soybean extract having hepatocellular protective effect and bone mineral density increasing effect and composition comprising same

๋ณธ ๋ฐœ๋ช…์€ ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ํšจ๊ณผ ๋ฐ ๊ณจ ๋ฐ€๋„ ์ฆ๊ฐ€ ํšจ๊ณผ๋ฅผ ๊ฐ–๋Š” ์ฟ ๋ฉ”์Šคํ…Œ๋กค ํ•จ์œ  ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ ๋ฐ ์ด๋ฅผ ํฌํ•จํ•˜๋Š” ์กฐ์„ฑ๋ฌผ์— ๊ด€ํ•œ ๊ฒƒ์ด๋‹ค. ๋ณด๋‹ค ์ƒ์„ธํ•˜๊ฒŒ๋Š” ์ž์—ฐ์‹ํ’ˆ ์ค‘์—์„œ ์ฒด๋‚ด ์ง€์งˆ ๋ฐ ๊ฐฑ๋…„๊ธฐ ์žฅ์•  ๊ฐœ์„ ํšจ๊ณผ๊ฐ€ ์žˆ๋Š” ์„ฑ๋ถ„์œผ๋กœ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์ด์šฉํ•˜์—ฌ ๊ฐฑ๋…„๊ธฐ ์žฅ์•  ์‹œ ๋‚˜ํƒ€๋‚˜๋Š” ๊ฐ„๋…์„ฑ์— ๋Œ€ํ•˜์—ฌ in vitro ํ•ญ์‚ฐํ™”ํ™œ์„ฑ๊ณผ ์„ธํฌ์ƒ์กด์œจ ๊ทธ๋ฆฌ๊ณ  ๋™๋ฌผ๋ชจ๋ธ์˜ ํ˜ˆ์•กํ•™์  ๊ฒ€์‚ฌ, MDA(malone dialdehyde), GST(glutathione S-transferase), GSH(ํ™˜์›ํ˜•๊ธ€๋ฃจํƒ€์น˜์˜จ), SOD(Superoxide dismutase), Catalase ๋“ฑ์˜ ์ธก์ •์„ ํ†ตํ•ด ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ํšจ๊ณผ๋ฅผ ๊ฒ€์ฆํ•˜๊ณ ์ž ํ•œ๋‹ค. ๋˜ํ•œ, ๋ณธ ๋ฐœ๋ช…์—์„œ๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์‹œ๋ฃŒ๋กœ ํ•˜์—ฌ in vivo ์‹คํ—˜์„ ํ†ตํ•ด ๊ฐฑ๋…„๊ธฐ ์žฅ์•  ์‹œ ๋‚˜ํƒ€๋‚  ์ˆ˜ ์žˆ๋Š” ๊ณจ ์†์‹ค์˜ ๊ฐœ์„ ํšจ๊ณผ๋ฅผ ๊ฒ€ํ† ํ•˜๊ธฐ ์œ„ํ•˜์—ฌ ํ˜ˆ์ค‘ ์นผ์Š˜๊ณผ ์ธ์˜ ๋†๋„, ๊ณจ ํ˜•์„ฑ ์ง€ํ‘œ์ธ ALP ํ™œ์„ฑ, ๊ณจ๋Ÿ‰(Bone mineral content)์„ ํ™•์ธํ•˜์˜€๊ณ , ํ•ด๋ฉด๊ณจ์˜ ๋‹ค์–‘ํ•œ ์ง€ํ‘œ๋ฅผ Micro-CT๋ฅผ ์ด์šฉํ•˜์—ฌ ์ธก์ •ํ•˜์˜€๋‹ค.The present invention relates to cousterol-containing germinated fermented soybean extract having a hepatocellular protective effect and an effect of increasing bone density and a composition comprising the same. More specifically, the in vitro antioxidant activity, cell viability, hematological test of animal models, MDA (malone) for the hepatotoxicity of menopausal disorders using germinated fermented soybean extract as a component that improves lipids and menopausal disorders in natural foods Dialdehyde), GST (glutathione S-transferase), GSH (reduced glutathione), SOD (Superoxide dismutase), and catalase were measured to verify hepatocellular protective effects. In the present invention, the germinated fermented soybean extract as a sample in vivo In order to examine the effect of improving bone loss in menopausal disorders, we examined the concentrations of calcium and phosphorus in blood, ALP activity, bone mineral content, and bone mineral content. It measured using.

๋ณธ ๋ฐœ๋ช…์˜ ๋ฐœํšจ๋ฐœ์•„์ฝฉ ์ถ”์ถœ๋ฌผ์€ ํ˜ˆ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ๊ณผ ํ…Œ์Šคํ† ์Šคํ…Œ๋ก  ๋†๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ•˜๋ฏ€๋กœ ์ธํ•ด ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์„ ์™„ํ™”์‹œํ‚ค๊ณ , in vitro ์™€ in vivo์‹คํ—˜์—์„œ ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ํšจ๊ณผ์™€, ๊ณจ์˜ ๋ฏธ๋„ค๋ž„ ํ•จ๋Ÿ‰์„ ์ฆ๊ฐ€์‹œ์ผœ ๊ณจ์˜ ๋ฐ€๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ•˜๊ณ , ํ•ด๋ฉด๊ณจ์˜ ๋‘๊ป˜์™€ ์ˆ˜๊ฐ€ ์ฆ๊ฐ€ํ•œ๋‹คFermented germinated soybean extract of the present invention significantly increases blood estrogen and testosterone concentrations, thereby relieving menopausal symptoms, and increasing bone mineral protection and increasing bone mineral content in vitro and in vivo. Increase, and the thickness and number of spongy bones increase

์ฝฉ๋ฅ˜์˜ ์ด์†Œํ”Œ๋ผ๋ณธ ์ค‘์˜ ๋‹ค์ด๋“œ์ œ์ธ(didzein)์— ์ฟ ๋ฉ”์Šคํ…Œ๋กค ์„ฑ๋ถ„์ด ๋“ค์–ด ์žˆ์œผ๋ฉฐ ์—ฌ์„ฑ ํ˜ธ๋ฅด๋ชฌ ๊ท ํ˜• ๋ฐ ํ•จ๋Ÿ‰์„ ์กฐ์ ˆํ•˜์—ฌ ์œ ๋ฐฉ์•”๊ณผ ์ž๊ถ์•”์„ ์˜ˆ๋ฐฉํ•ด ์ฃผ๋ฉฐ ํ๊ฒฝ๊ธฐ๋ฅผ ๋Šฆ์ถ”๊ฑฐ๋‚˜ ์ด์™€ ๊ด€๋ จ๋œ ์—ฌ๋Ÿฌ ์ฆ์ƒ์„ ์™„ํ™”์‹œํ‚จ๋‹ค.Didzein in legumes isoflavones contain cumesterol, which regulates female hormone balance and content to prevent breast and uterine cancer, slowing menopause and alleviating many of the symptoms associated with it.

์ธ๊ตฌ์˜ ๊ณ ๋ นํ™”๋Š” ์ „ ์„ธ๊ณ„์ ์œผ๋กœ ๋งค์šฐ ๊ธ‰์†ํ•˜๊ฒŒ ์ง„ํ–‰๋˜๊ณ  ์žˆ์œผ๋ฉฐ, ํŠนํžˆ, ์šฐ๋ฆฌ๋‚˜๋ผ์˜ ๊ฒฝ์šฐ ๊ณ ๋ นํ™” ์ง„์ „์†๋„๊ฐ€ ๋‹ค๋ฅธ ์„ ์ง„๊ตญ๊ณผ ๋น„๊ตํ•˜์—ฌ ์ง„ํ–‰์†๋„๊ฐ€ ๋งค์šฐ ๋นจ๋ผ ํฐ ๋ฌธ์ œ๊ฐ€ ๋˜๊ณ  ์žˆ๋‹ค. ๊ฒฝ์ œ์„ฑ์žฅ์— ๋”ฐ๋ฅธ ๊ฑด๊ฐ• ์ฆ์ง„์˜ ๊ฒฐ๊ณผ๋กœ ์ธํ•œ ์šฐ๋ฆฌ๋‚˜๋ผ ์—ฌ์„ฑ์˜ ํ‰๊ท ์ˆ˜๋ช…์˜ ์—ฐ์žฅ๊ณผ ์ง€์  ์ˆ˜์ค€์˜ ํ–ฅ์ƒ ๋“ฑ์œผ๋กœ ์—ฌ์„ฑ์˜ ํ๊ฒฝ๊ธฐ์™€ ํ๊ฒฝ ํ›„์˜ ์‚ถ์˜ ์งˆ ํ–ฅ์ƒ๊ณผ ๊ฑด๊ฐ• ์ฆ์ง„์— ๋Œ€ํ•œ ๊ด€์‹ฌ ๋˜ํ•œ ์ ์  ๊ณ ์กฐ๋˜๊ณ  ์žˆ๋‹ค. ํ๊ฒฝ์˜ ์ฃผ์š” ์ฆ์ƒ์œผ๋กœ๋Š” ์•ˆ๋ฉดํ™์กฐ, ๋ถˆ์•ˆ์ฆ, ์šฐ์šธ, ์‹ ๊ฒฝ๊ณผ๋ฏผ. ์‹ฌํ˜ˆ๊ด€์งˆํ™˜, ๊ณจ๋‹ค๊ณต์ฆ ๋ฐ ๊ธฐ์–ต๋ ฅ ๊ฐํ‡ด ๋“ฑ์˜ ์ฆ์ƒ์ด ๋Œ€ํ‘œ์ ์ด๋ฉฐ ์ด๋Ÿฌํ•œ ์ฆ์ƒ์˜ ์›์ธ์€ ์—์ŠคํŠธ๋กœ๊ฒ์˜ ์ƒ์‚ฐ๊ณผ ๋ถ„๋น„๊ฐ€ ์ค‘๋‹จ๋˜์–ด ๋‚˜ํƒ€๋‚œ๋‹ค๊ณ  ์•Œ๋ ค์ ธ ์žˆ๋‹ค. ์ด์˜ ์˜ˆ๋ฐฉ์ฑ…์œผ๋กœ ํ˜ธ๋ฅด๋ชฌ ๋Œ€์ฒด์š”๋ฒ•(estrogen replacement therapy)์ด ์ œ์‹œ๋˜๊ณ  ์žˆ์œผ๋‚˜ ์‚ฌ์šฉ๋ฒ•, ์šฉ๋Ÿ‰, ์•ˆ์ „์„ฑ ๋“ฑ์— ๋Œ€ํ•œ ๊ฒฐ๊ณผ๋Š” ์—†๋‹ค.The aging of the population is progressing very rapidly all over the world, and especially in Korea, the progress of aging is very fast compared to other developed countries. As women's average life expectancy is increased and the intellectual level is improved as a result of the health promotion resulting from economic growth, interest in women's menopause and postmenopausal quality of life and health improvement is also increasing. The main symptoms of menopause include hot flashes, anxiety, depression and nervousness. Symptoms such as cardiovascular disease, osteoporosis and memory loss are known. The cause of these symptoms is known to be caused by interrupted production and secretion of estrogen. As a preventive measure, hormone replacement therapy has been proposed, but there are no results on usage, dose, and safety.

์ฒœ์—ฐ ์‹ํ’ˆ ์ค‘์˜ ์‹๋ฌผ์„ฑ ์—์ŠคํŠธ๋กœ๊ฒ(Phytoestrogen)์€ ์ƒ์‹ ํ˜ธ๋ฅด๋ชฌ ์œ ์‚ฌ๋ฌผ์งˆ๋กœ ํ๊ฒฝ๊ธฐ ์ฆ์ƒ ๋ฐ ๊ณจ๋‹ค๊ณต์ฆ ์˜ˆ๋ฐฉ ๊ฐ€๋Šฅ์„ฑ์ด ์ฃผ๋ชฉ ๋ฐ›๊ธฐ ์‹œ์ž‘ํ•˜๋ฉด์„œ ํ๊ฒฝ๊ธฐ ์ดํ›„์˜ ์—ฌ์„ฑ์—๊ฒŒ estrogen ๋Œ€์ฒด์ž‘์šฉ์„ ํ•  ์ˆ˜ ์žˆ๋‹ค๋Š” ๊ฒƒ์ด ๋งŽ์ด ์—ฐ๊ตฌ, ๋ณด๊ณ ๋˜๊ณ  ์žˆ๋‹ค. ํ˜„์žฌ ํ๊ฒฝ๊ธฐ ์ฆ์ƒ ์น˜๋ฃŒ์— ํ•ฉ์„ฑ estrogen ๋Œ€์ฒด ์š”๋ฒ•์ด ์ด์šฉ๋˜๋ฉด์„œ, ๋Œ€๋ถ€๋ถ„์˜ ์—ฌ์„ฑ๋“ค์€ ์žฌ ์›”๊ฒฝ ํ˜„์ƒ, ์œ ๋ฐฉ์˜ ์—ฐํ™” ๋ฐ ์ฒด์ค‘์ฆ๊ฐ€ ๋“ฑ์˜ ๋ถ€์ž‘์šฉ์„ ๊ฒฝํ—˜ํ•˜๊ณ  ์žˆ๊ณ , ์ฒœ์—ฐ ์‹ํ’ˆ ์†์—์„œ์˜ ์‹๋ฌผ์„ฑ estrogen ์œ ์‚ฌ๋ฌผ์งˆ์„ ๊ฐ€์ง„ ์‹ํ’ˆ์„ญ์ทจ๋ฅผ ํ†ตํ•œ ํ๊ฒฝ๊ธฐ ์ฆ์ƒ ์™„ํ™”๋Š” ๋ถ€์ž‘์šฉ ํ•ด์†Œ์— ๋งŽ์€ ๋„์›€์„ ์ฃผ๋ฆฌ๋ผ ์ƒ๊ฐํ•œ๋‹ค.Phytoestrogen, a natural food product, is a reproductive hormone-like substance that is beginning to attract attention for the prevention of menopausal symptoms and osteoporosis, and has been studied and reported to be able to substitute estrogen in postmenopausal women. As synthetic estrogen replacement therapy is currently used to treat menopausal symptoms, most women experience side effects such as remenstruation, breast softening and weight gain, and food intake with plant-based estrogen-like substances in natural foods. Relieving menopausal symptoms will help to alleviate the side effects.

์‹๋ฌผ์„ฑ ์—์ŠคํŠธ๋กœ๊ฒ์€ ์ด์†Œํ”Œ๋ผ๋ณธ๊ณผ ๋ฆฌ๊ทธ๋‚œ์„ ํฌํ•จํ•˜๋ฉฐ, ์ฝฉ, ๊ณผ์ผ, ์ฑ„์†Œ, ๋•…์ฝฉ, ๊ณก๋ฌผ ๋“ฑ์˜ ์‹๋ฌผ์—์„œ ์ž์—ฐ์ ์œผ๋กœ ๋ฐœ์ƒํ•œ ์—์ŠคํŠธ๋กœ๊ฒ๊ณผ ์œ ์‚ฌํ•œ ํ™”ํ•ฉ๋ฌผ์ด๋‹ค. ์ด์†Œํ”Œ๋ผ๋ณธ์€ ์ฒด๋‚ด ์—์ŠคํŠธ๋กœ๊ฒ ์ˆ˜์šฉ์ฒด์™€ ์•ฝํ•˜๊ฒŒ ๊ฒฐํ•ฉํ•˜์—ฌ ๋‹ค์–‘ํ•œ ์ƒ๋ฆฌ ํšจ๊ณผ๋ฅผ ๊ฐ€์ง€๋Š” ์‹๋ฌผ์„ฑ ์—์ŠคํŠธ๋กœ๊ฒ์˜ ์ผ์ข…์œผ๋กœ ์ฃผ๋ชฉ ๋ฐ›๊ณ  ์žˆ๋‹ค. ์ฝฉ์—์„œ ์ถ”์ถœํ•œ ์ด์†Œํ”Œ๋ผ๋ณธ์€ ์‹๋ฌผ ๋‚ด์—์„œ๋Š” ์ฃผ๋กœ glycoside๋กœ ์กด์žฌํ•˜๋ฉฐ, ์ธ์ฒด ๋‚ด๋กœ ํก์ˆ˜๋˜๋ฉด์„œ glycosidase์— ์˜ํ•ด ์•„๊ธ€๋ฆฌ์ฝ˜ ํ˜•ํƒœ์˜ ์ œ๋‹ˆ์Šคํ…Œ์ธ๊ณผ ๋‹ค์ด๋“œ์ œ์ธ์œผ๋กœ ๋ณ€ํ™”๋˜์–ด estrogenic property๋ฅผ ๊ฐ–๊ฒŒ ๋œ๋‹ค.Phytoestrogens include isoflavones and lignans and are compounds similar to those naturally occurring in plants such as beans, fruits, vegetables, peanuts and grains. Isoflavones are attracting attention as a kind of phytoestrogens that weakly bind to estrogen receptors in the body and have various physiological effects. Isoflavones extracted from soybeans exist mainly as glycosides in plants, and are absorbed into the human body to be converted into aglycone forms of genistein and dyedzein by glycosidase and have estrogenic properties.

์—ฌ์„ฑ์˜ ๊ฑด๊ฐ•๊ณผ ๊ด€๋ จํ•˜์—ฌ ์ด์†Œํ”Œ๋ผ๋ณธ์˜ ์„ญ์ทจ๋Š” ๊ธฐ์กด์˜ ์—์ŠคํŠธ๋กœ๊ฒ ์น˜๋ฃŒ์ œ์™€ ๋‹ฌ๋ฆฌ ๋ถ€์ž‘์šฉ ์—†์ด ํ๊ฒฝ๊ธฐ ์ฆ์ƒ์„ ์™„ํ™”์‹œํ‚ค๊ณ , ํ˜ˆ์ฒญ ์ง€์งˆ ๋†๋„ ๋ฐ ๊ณ ์ฝœ๋ ˆ์Šคํ…Œ๋กค ํ˜ˆ์ฆ์„ ๋ณด์ด๋Š” ์—ฌ์„ฑ์—์„œ ์‹ฌํ˜ˆ๊ด€๊ณ„ ์œ„ํ—˜๋„์™€ ํ˜ˆ์ค‘ ์ง€์งˆ๊ณผ ์‚ฐํ™”๋ฌผ์„ ๊ฐ์†Œ์‹œํ‚ค๋Š” ๊ฒƒ์œผ๋กœ ๋ณด๊ณ ๋œ ๋ฐ” ์žˆ๋‹ค.Concerning women's health, ingestion of isoflavones has been reported to alleviate menopausal symptoms without side effects, unlike conventional estrogen therapy, and to reduce cardiovascular risk and blood lipids and oxides in women with serum lipid levels and hypercholesterolemia. There is a bar.

๋Œ€ํ•œ๋ฏผ๊ตญ ๊ณต๊ฐœํŠนํ—ˆ๊ณต๋ณด ์ œ10-2014-0053471(์ฟ ๋ฉ”์ŠคํŠธ๋กค์„ ํฌํ•จํ•˜๋Š” ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์œ ํšจ์„ฑ๋ถ„์œผ๋กœ ํฌํ•จํ•˜๋Š” ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์˜ ์˜ˆ๋ฐฉ ๋ฐ ์น˜๋ฃŒ์šฉ ์กฐ์„ฑ๋ฌผ)์€ ์ฟ ๋ฉ”์ŠคํŠธ๋กค์„ ํฌํ•จํ•˜๋Š” ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์œ ํšจ์„ฑ๋ถ„์œผ๋กœ ํฌํ•จํ•˜๋Š” ๊ฐฑ๋…„๊ธฐ ์งˆํ™˜์˜ ์˜ˆ๋ฐฉ ๋“ฑ์— ๋Œ€ํ•œ ์กฐ์„ฑ๋ฌผ์— ๊ด€ํ•œ ์„ธํฌ์‹คํ—˜(in vitro)์˜ ์‹คํ—˜ ๊ฒฐ๊ณผ์ด๋ฉฐ, ๋ณธ์›๋ฐœ๋ช…์—์„œ ์ƒ๊ธฐ์˜ ์„ธํฌ์‹คํ—˜์„ ํ™•์ธํ•˜๊ณ ์ž ๋™๋ฌผ์‹คํ—˜(in-vivo)์„ ํ†ตํ•ด ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ ๊ฐœ์„  ๋ฐ ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ํšจ๊ณผ ๋“ฑ์„ ํ™•์ธํ•˜๊ณ ์ž ๊ฐœ๋Ÿ‰ ๋ฐ ๊ฐœ์„ ๋œ ๋ฐœ์•„๋ฐœํšจ์ฝฉ์ถ”์ถœ๋ฌผ๋กœ ์ง„๋ณด๋œ ๋ฐฉ๋ฒ•๊ณผ ๊ฒฐ๊ณผ๋ฅผ ์–ป๊ณ ์ž ํ•˜๋Š” ๋ชฉ์ ์ž…๋‹ˆ๋‹ค.Republic of Korea Patent Publication No. 10-2014-0053471 (composition for the prevention and treatment of menopausal symptoms comprising a soybean extract containing cumestrol as an active ingredient) a menopause containing a soybean extract containing cumestrol as an active ingredient It is the experimental result of the cell experiment (in vitro) on the composition for the prevention of diseases, and to improve the menopausal symptoms improvement and liver cell protection effect through the animal experiment (in-vivo) to confirm the cell experiment in the present invention. And improved germinated fermented soybean extract to achieve advanced methods and results.

{์„ ํ–‰๊ธฐ์ˆ ๋ฌธํ—Œ}{Prior art technical literature}

(ํŠนํ—ˆ๋ฌธํ—Œ 1) ํ•œ๊ตญํŠนํ—ˆ ๊ณต๊ฐœ๋ฒˆํ˜ธ 10-2014-0053471(Patent Document 1) Korean Patent Publication No. 10-2014-0053471

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 1)1. Starzec, J,J, and D.F. Berger, Effects of stress and ovariectomy on the plasma cholesterol, serum triglyceride, and aortic cholesterol levels of female rats, Physiol Behav. 1986.37(1): p.99-104.(Non-Patent Document 1) 1. Starzec, J, J, and D.F. Berger, Effects of stress and ovariectomy on the plasma cholesterol, serum triglyceride, and aortic cholesterol levels of female rats, Physiol Behav. 1986.37 (1): p. 99-104.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 2)2. Ettinger, B., Overview of estrogen replacement therapy a historical perspective. Proc Soc Exp Biol Med. 217(1): p.2-5.(Non-Patent Document 2) 2. Ettinger, B., Overview of estrogen replacement therapy a historical perspective. Proc Soc Exp Biol Med. 217 (1): p. 2-5.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 3)3. Baker, M.E.,K.L. Medlock, and D.M. Sheehan, Flavonoids inhibit estrogen binding to rat alpha-fetoprotein. Proc Soc Exp Biol Med. 1998. 217(3): p.317-21.(Non-Patent Document 3) 3. Baker, M.E., K.L. Medlock, and D.M. Sheehan, Flavonoids inhibit estrogen binding to rat alpha-fetoprotein. Proc Soc Exp Biol Med. 1998.217 (3): p.317-21.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 4)4. Boker, L.K., et al., Intake of Dietary Phytoestrogens by Dutch women. J Nutr. 2002. 132(6): p.1319-28.(Non-Patent Document 4) 4. Boker, L.K., et al., Intake of Dietary Phytoestrogens by Dutch women. J Nutr. 2002. 132 (6): p. 1319-28.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 5)5. Morito, K., et al., Interaction of Phytoestrogens with estrogen receptors alpha and beta, Biol Pharm Bull. 2001. 24(4); p.6-11.(Non-Patent Document 5) 5. Morito, K., et al., Interaction of Phytoestrogens with estrogen receptors alpha and beta, Biol Pharm Bull. 2001. 24 (4); p.6-11.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 6)6. Albertazzi, P., et al., The effect of dietary soy supplementation on hot flushes, Obstet Gynecol. 1988. 91(3): p.389-94.(Non-Patent Document 6) 6. Albertazzi, P., et al., The effect of dietary soy supplementation on hot flushes, Obstet Gynecol. 1988. 91 (3): p. 389-94.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 7)7. Han, K,K., et al., Benefits of soy isoflavone therapeutic regimen on menopausal symptoms, Obstet Gynecol. 2002. 99(3): p.389-94. (Non-Patent Document 7) 7. Han, K, K., et al., Benefits of soy isoflavone therapeutic regimen on menopausal symptoms, Obstet Gynecol. 2002. 99 (3): p. 389-94.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 8)8. Petri Nahas, E., et al., Benefits of soy germ isoflavones in postmenopausal women with contraindication for conventional hormone replacement therapy, Maturitas. 2004. 48(4): p.372-80.(Non-Patent Document 8) 8. Petri Nahas, E., et al., Benefits of soy germ isoflavones in postmenopausal women with contraindication for conventional hormone replacement therapy, Maturitas. 2004. 48 (4): p.372-80.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 9)9. Kaari, C., ey al., Randomized clinical trial comparing conjugated equine estrogens and isoflavones in postmenopausal women: a pilot study, Maturitus. 2006. 53(1): p.49-58.(Non-Patent Document 9) 9. Kaari, C., ey al., Randomized clinical trial comparing conjugated equine estrogens and isoflavones in postmenopausal women: a pilot study, Maturitus. 2006. 53 (1): p. 49-58.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 10)10. Cheng, G., et al., Isoflavone treatment for acute menopausal symptoms, Menopause. 2007.14(3Pt1):p.468-73.(Non-Patent Document 10) Cheng, G., et al., Isoflavone treatment for acute menopausal symptoms, Menopause. 2007.14 (3Pt1): p.468-73.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 11) DJ., et al., Effect of soy protein foods on low-density lipoprotein oxidation and ex vivo sex hormone receptor activity - - a controlled crossover trial. Metabolism. 2000.49(4):p.537-43.(Non-Patent Document 11) DJ., Et al., Effect of soy protein foods on low-density lipoprotein oxidation and ex vivo sex hormone receptor activity--a controlled crossover trial. Metabolism. 2000.49 (4): p.537-43.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 12)12. Kotsopoulos, D., et al., The effects of soy protein containing phytoesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.55(21):p.8589-95.(Nonpatent Document 12) Kotsopoulos, D., et al., The effects of soy protein containing phytoesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.55 (21): p. 8589-95.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 13)13. Wangen, K.E., et al., Soy Isoflavones improve plasma lipids in normocholesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.73(2):p.225-31.(Non-Patent Document 13) 13. Wangen, K.E., et al., Soy Isoflavones improve plasma lipids in normocholesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.73 (2): p.225-31.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 14)14. Feng, S., et al., Fungal-stressed germination of black soybeans leads to generation of oxooctadecadienoic acids in addition to glyceollins, J Agric Food Chem. 2007.55(21):p.8589-95.(Non-Patent Document 14) 14. Feng, S., et al., Fungal-stressed germination of black soybeans leads to generation of oxooctadecadienoic acids in addition to glyceollins, J Agric Food Chem. 2007.55 (21): p.8589-95.

(๋น„ํŠนํ—ˆ๋ฌธํ—Œ 15)15. Rosalki, S.B. and A.Y.Foo. Two new methods for seperating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma, Cli Chem. 1984. 30(7): p.1182-6.(Non-Patent Document 15) 15. Rosalki, S.B. and A.Y.Foo. Two new methods for seperating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma, Cli Chem. 1984. 30 (7): p.1182-6.

๋ณธ ๋ฐœ๋ช…์€ ๋ฐœํšจ๋ฐœ์•„์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์ด์šฉํ•˜์—ฌ ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์„ ์™„ํ™”์‹œํ‚ค๊ณ , ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ํšจ๊ณผ์™€, ๊ณจ์˜ ๋ฏธ๋„ค๋ž„ ํ•จ๋Ÿ‰์„ ์ฆ๊ฐ€์‹œํ‚ค๋Š”๋ฐ ์žˆ๋‹ค.The present invention uses fermented germinated soybean extract to relieve menopausal symptoms, to protect the liver cells, and to increase the mineral content of bone.

๊ฒฝ์ œ์„ฑ์žฅ์— ๋”ฐ๋ฅธ ๊ฑด๊ฐ• ์ฆ์ง„์˜ ๊ฒฐ๊ณผ๋กœ ์ธํ•œ ์šฐ๋ฆฌ๋‚˜๋ผ ์—ฌ์„ฑ์˜ ํ‰๊ท ์ˆ˜๋ช…์˜ ์—ฐ์žฅ๊ณผ ์ง€์  ์ˆ˜์ค€์˜ ํ–ฅ์ƒ ๋“ฑ์œผ๋กœ ์—ฌ์„ฑ์˜ ํ๊ฒฝ๊ธฐ์™€ ํ๊ฒฝ ํ›„์˜ ์‚ถ์˜ ์งˆ ํ–ฅ์ƒ๊ณผ ๊ฑด๊ฐ• ์ฆ์ง„์— ๋Œ€ํ•œ ๊ด€์‹ฌ ๋˜ํ•œ ์ ์  ๊ณ ์กฐ๋˜๊ณ  ์žˆ๋‹ค. ํ๊ฒฝ์˜ ์ฃผ์š” ์ฆ์ƒ์œผ๋กœ๋Š” ์•ˆ๋ฉดํ™์กฐ, ๋ถˆ์•ˆ์ฆ, ์šฐ์šธ, ์‹ ๊ฒฝ๊ณผ๋ฏผ. ์‹ฌํ˜ˆ๊ด€์งˆํ™˜, ๊ณจ๋‹ค๊ณต์ฆ ๋ฐ ๊ธฐ์–ต๋ ฅ ๊ฐํ‡ด ๋“ฑ์˜ ์ฆ์ƒ์ด ๋Œ€ํ‘œ์ ์ด๋ฉฐ ์ด๋Ÿฌํ•œ ์ฆ์ƒ์˜ ์›์ธ์€ ์—์ŠคํŠธ๋กœ๊ฒ์˜ ์ƒ์‚ฐ๊ณผ ๋ถ„๋น„๊ฐ€ ์ค‘๋‹จ๋˜์–ด ๋‚˜ํƒ€๋‚œ๋‹ค๊ณ  ์•Œ๋ ค์ ธ ์žˆ๋‹ค. ์ด์˜ ์˜ˆ๋ฐฉ์ฑ…์œผ๋กœ ํ˜ธ๋ฅด๋ชฌ ๋Œ€์ฒด์š”๋ฒ•(estrogen replacement therapy)์ด ์ œ์‹œ๋˜๊ณ  ์žˆ์œผ๋‚˜ ์‚ฌ์šฉ๋ฒ•, ์šฉ๋Ÿ‰, ์•ˆ์ „์„ฑ ๋“ฑ์— ๋Œ€ํ•œ ๊ฒฐ๊ณผ๋Š” ์—†๋‹ค. ๋”์šฑ์ด ํ๊ฒฝ๊ธฐ์˜ ์—ฌ์„ฑ์— ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ํšจ๊ณผ ๋ฐ ๊ณจ ๋ฐ€๋„ ์ฆ๊ฐ€ ํšจ๊ณผ๋ฅผ ๊ฐ–๋Š” ๋งˆ๋•…ํ•œ ์•ฝํ’ˆ์ด๋‚˜ ๊ฑด๊ฐ•์‹ํ’ˆ์ด ๋ถ€์กฑํ•œ ์‹ค์ •์ด๋‹ค.As women's average life expectancy is increased and the intellectual level is improved as a result of the health promotion resulting from economic growth, interest in women's menopause and postmenopausal quality of life and health improvement is also increasing. The main symptoms of menopause include hot flashes, anxiety, depression and nervousness. Symptoms such as cardiovascular disease, osteoporosis and memory loss are known. The cause of these symptoms is known to be caused by interrupted production and secretion of estrogen. As a preventive measure, hormone replacement therapy has been proposed, but there are no results on usage, dose, and safety. In addition, there is a lack of appropriate drugs or health foods that have hepatoprotective effect and bone mineral density increase effect in postmenopausal women.

๋ณธ ๋ฐœ๋ช…์€ ํ๊ฒฝ๊ธฐ์˜ ์—ฌ์„ฑ์— ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ๋ฐ ๊ณจ ๋ฐ€๋„ ์ฆ๊ฐ€ ํšจ๊ณผ๋ฅผ ๊ฐ–๋Š” ์ฟ ๋ฉ”์Šคํ…Œ๋กค ํ•จ์œ  ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ ๋ฐ ์ด๋ฅผ ํฌํ•จํ•˜๋Š” ์กฐ์„ฑ๋ฌผ์— ๊ด€ํ•œ ๊ฒƒ์ด๋‹ค. ์ƒ๊ธฐ์˜ ์ฟ ๋ฉ”์Šคํ…Œ๋กค์„ ํ•จ์œ ํ•œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์ด์šฉํ•˜์—ฌ ํ๊ฒฝ๊ธฐ์˜ ์—ฌ์„ฑ์— ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ๋ฐ ๊ณจ ๋ฐ€๋„ ์ฆ๊ฐ€ ํšจ๊ณผ๋ฅผ ๊ฒ€์ฆํ•˜๊ณ ์ž ํ•œ๋‹ค. ๋˜ํ•œ, ๋ณธ ๋ฐœ๋ช…์—์„œ๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์‹œ๋ฃŒ๋กœ ํ•˜์—ฌ in vivo ์‹คํ—˜์„ ํ†ตํ•ด ๊ฐฑ๋…„๊ธฐ ์žฅ์•  ์‹œ ๋‚˜ํƒ€๋‚  ์ˆ˜ ์žˆ๋Š” ํ๊ฒฝ๊ธฐ์˜ ์—ฌ์„ฑ์— ๊ฐ„์„ธํฌ ๋ณดํ˜ธ ๋ฐ ๊ณจ ์†์‹ค์˜ ๊ฐœ์„ ํšจ๊ณผ๋ฅผ ๊ฒ€ํ† ํ•˜๊ธฐ ์œ„ํ•˜์—ฌ ํ˜ˆ์ค‘ ์นผ์Š˜๊ณผ ์ธ์˜ ๋†๋„, ๊ณจ ํ˜•์„ฑ ์ง€ํ‘œ์ธ ALP ํ™œ์„ฑ, ๊ณจ๋Ÿ‰(Bone mineral content)์„ ํ™•์ธํ•˜์˜€๊ณ , ํ•ด๋ฉด๊ณจ์˜ ๋‹ค์–‘ํ•œ ์ง€ํ‘œ๋ฅผ Micro-CT๋ฅผ ์ด์šฉํ•˜์—ฌ ์ธก์ •ํ•˜์˜€๋‹ค.The present invention relates to a cousterol-containing germinated fermented soybean extract having an effect of protecting liver cells and increasing bone density in postmenopausal women and a composition comprising the same. The germinated fermented soybean extract containing cumesterol to verify the effect of hepatocellular protection and bone density increase in postmenopausal women. In the present invention, the germinated fermented soybean extract as a sample in vivo In order to examine the effect of improving hepatocellular protection and bone loss in postmenopausal women during menopausal disorders, we examined blood calcium and phosphorus concentrations, ALP activity, bone mineral content, and bone mineral content. Various indicators were measured using Micro-CT.

ํ˜ˆ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ•˜๋Š” ๊ฒƒ์„ ํ†ตํ•ด ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ isoflavone์ด ์—์ŠคํŠธ๋กœ๊ฒ์˜ ์—ญํ• ์„ ๋Œ€์ฒดํ•  ์ˆ˜ ์žˆ๊ณ  ์ด๋กœ ์ธํ•ด ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์ด ์™„ํ™”๋  ๊ฒƒ์„ ๊ธฐ๋Œ€ํ•  ์ˆ˜ ์žˆ๋‹ค. ํ…Œ์Šคํ† ์Šคํ…Œ๋ก  ๋†๋„๋Š” ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ์œ ์˜์ ์œผ๋กœ (p < 0.05) ์ฆ๊ฐ€ํ•œ ๊ฒฐ๊ณผ์ž„์„ ํ™•์ธํ•˜์˜€๋‹ค.Significant increases in blood estrogen levels can be expected to replace the role of estrogens in germinated fermented soybeans, thereby alleviating menopausal symptoms. Testosterone concentration was found to be significantly increased (p <0.05) compared to the ovarian control group.

๋ณธ ๋ฐœ๋ช…์€ ๊ฐฑ๋…„๊ธฐ ์žฅ์•  ์‹œ ๋‚˜ํƒ€๋‚˜๋Š” ๊ฐ„๋…์„ฑ์— ๋Œ€ํ•˜์—ฌ in vitro ํ•ญ์‚ฐํ™”ํ™œ์„ฑ๊ณผ ์„ธํฌ์ƒ์กด์œจ ๊ทธ๋ฆฌ๊ณ  ๋™๋ฌผ๋ชจ๋ธ์˜ ํ˜ˆ์•กํ•™์  ๊ฒ€์‚ฌ, MDA, GST, GSH, SOD, CAT๋“ฑ์˜ ์ธก์ •์„ ํ†ตํ•ด ๊ฐ„์„ธํฌ ๋ณดํ˜ธ์— ํšจ๊ณผ์ ์ž„์„ ์•Œ ์ˆ˜ ์žˆ์—ˆ๋‹ค. ๋˜ํ•œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ํˆฌ์—ฌ๊ฐ€ ๊ณจ์˜ ๋ฏธ๋„ค๋ž„ ํ•จ๋Ÿ‰์„ ์ฆ๊ฐ€์‹œํ‚ค๋Š” ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค. ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ์ฅ์—์„œ ๊ณจ์˜ ๋ฐ€๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ–ˆ๊ณ , ํ•ด๋ฉด๊ณจ์˜ ๋‘๊ป˜์™€ ์ˆ˜๊ฐ€ ์ฆ๊ฐ€ํ•˜์˜€๋‹ค๋Š” ๊ฒƒ์„ ์˜๋ฏธํ•œ๋‹ค. ํ˜ˆ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ•˜๋Š” ๊ฒƒ์„ ํ†ตํ•ด ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ isoflavone์ด ์—์ŠคํŠธ๋กœ๊ฒ์˜ ์—ญํ• ์„ ๋Œ€์ฒดํ•  ์ˆ˜ ์žˆ๊ณ  ์ด๋กœ ์ธํ•ด ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์ด ์™„ํ™”๋  ๊ฒƒ์„ ๊ธฐ๋Œ€ํ•  ์ˆ˜ ์žˆ๋‹ค. ํŠนํžˆ, ํ˜ˆ์•ก ๋‚ด ์ค‘์„ฑ์ง€๋ฐฉ์˜ ๋†๋„์™€ ํฌ๋„๋‹น์˜ ๋†๋„๊ฐ€ ๊ฐ์†Œํ•˜๋Š” ๊ฒฝํ–ฅ์„ ๊ฐ–๋Š” ๊ฒƒ์œผ๋กœ ๋ณด์•„, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ํˆฌ์—ฌ๊ฐ€ ๊ฐฑ๋…„๊ธฐ ๋น„๋งŒ ๊ฐœ์„ ์— ๋„์›€์„ ์ค„ ๊ฒƒ์œผ๋กœ ์˜ˆ์ƒ๋˜๊ณ , ๊ณ ๋†๋„์˜ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ํˆฌ์—ฌ๊ฐ€ ์—์ŠคํŠธ๋กœ๊ฒ ํˆฌ์—ฌ์™€ ๋น„์Šทํ•œ ์ˆ˜์ค€์œผ๋กœ ๊ณจ๋‹ค๊ณต์ฆ์˜ ์˜ˆ๋ฐฉํšจ๊ณผ๊ฐ€ ์žˆ๋‹ค๋Š” ๊ฒƒ์„ ํ™•์ธํ–ˆ๋‹ค. ์ธ๊ฐ„์˜ ๋‹ค์–‘ํ•œ ์งˆ๋ณ‘์˜ ์›์ธ์œผ๋กœ ์ง€๋ชฉ๋˜๋Š” ์‚ฐํ™”์  ์ŠคํŠธ๋ ˆ์Šค์— ๋Œ€ํ•œ ์ƒ์ฒด์˜ ์‚ฐํ™”์  ์†์ƒ์€ catalase, SOD, GSH, GST์™€ ๊ฐ™์€ ํ•ญ์‚ฐํ™” ํšจ์†Œ๋“ค์ด ํ•„์š”ํ•˜๋‹ค. ์ด์— ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ์ด์šฉํ•˜์—ฌ ์ธ์œ„์  ๊ฐ„ ์†์ƒ์„ ๊ฐ€์ง„ ๋™๋ฌผ๋ชจ๋ธ์„ ๊ฐ€์ง€๊ณ  ๊ฐ„ํšจ์†Œ ํ•ญ์‚ฐํ™” ํ™œ์„ฑ์„ ์ธก์ •ํ•œ ๊ฒฐ๊ณผ SOD, GSH, GST, MDA๋“ฑ t-BHP๋งŒ ํˆฌ์—ฌํ•œ ๊ฐ„์†์ƒ ๋ชจ๋ธ์— ๋Œ€๋น„ํ•˜์—ฌ ์œ ์˜์ ์œผ๋กœ ๊ฐ์†Œ ๋ฐ ์ฆ๊ฐ€๋ฅผ ๋‚˜ํƒ€๋ƒˆ๋‹ค. ์ด์— ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์€ ์‚ฐํ™”์  ์ŠคํŠธ๋ ˆ์Šค์— ๋Œ€ํ•œ ๊ฐ„ ๋ณดํ˜ธ ํšจ๊ณผ๊ฐ€ ์žˆ์Œ์„ ์•Œ ์ˆ˜ ์žˆ์—ˆ๋‹ค.The present invention was found to be effective for the protection of hepatocytes by measuring in vitro antioxidant activity, cell viability and hematological tests, MDA, GST, GSH, SOD, CAT, etc. for hepatotoxicity in menopausal disorders. It was also confirmed that the administration of germinated fermented soybeans increased the mineral content of bone. Bone density was significantly increased in the rats treated with germinated fermented soybean, and the thickness and number of spongy bones were increased. Significant increases in blood estrogen levels can be expected to replace the role of estrogens in germinated fermented soybeans, thereby alleviating menopausal symptoms. In particular, since the concentration of triglycerides and glucose in blood tend to decrease, the administration of germinated fermented soybean is expected to help improve menopausal obesity, and the high concentration of germinated fermented soybean is similar to that of estrogen. It was confirmed that there is a preventive effect of osteoporosis. The oxidative damage of the body to oxidative stress, which is the cause of various diseases in humans, requires antioxidant enzymes such as catalase, SOD, GSH, and GST. As a result of measuring liver enzyme antioxidant activity using an animal model with artificial liver damage using germinated fermented soybean, it was significantly reduced and increased compared to liver damage model administered only t- BHP such as SOD, GSH, GST, and MDA. Indicated. The germinated fermented soybean was found to have a liver protective effect against oxidative stress.

๋„ 1์€ ์—์ŠคํŠธ๋กœ๊ฒ์„ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ ์ฒด์ค‘ ์ฆ๊ฐ€๋Ÿ‰์„ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 1 shows the weight gain of the group receiving the estrogen.

๋„ 2๋Š” ํ˜ˆ์ฒญ ์ง€์งˆ, ์ฝœ๋ ˆ์Šคํ…Œ๋กค ๋ฐ ํฌ๋„๋‹น ๋†๋„๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹คFigure 2 shows serum lipids, cholesterol and glucose concentrations

๋„ 3์€ ํ˜ธ๋ฅด๋ชฌ์˜ ๋ณ€ํ™”๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.3 shows the change in hormone.

๋„ 4๋Š” ๊ณจ๋Œ€์‚ฌ ์ง€ํ‘œ ์ธก์ •๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 4 shows the bone metabolic index measurement results.

๋„ 5๋Š” ๊ณจ์—ผ๋Ÿ‰ ์ธก์ •๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 5 shows the results of bone salt measurement.

๋„ 6์€ ๊ณจ ๋ฏธ์„ธ ๊ตฌ์กฐ ์ง€ํ‘œ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.6 shows bone microstructural indicators.

๋„ 7์€ Micro-CT๋ฅผ ์ด์šฉํ•˜์—ฌ ์ดฌ์˜ํ•œ 2D์ด๋ฏธ์ง€๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 7 shows a 2D image taken by using the Micro-CT.

๋„ 8์€ 70% ์—ํƒ„์˜ฌ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ sep-pak์„ ์ด์šฉํ•œ ๋ถ„๋ฆฌ๊ณผ์ •์„ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 8 shows the separation process using sep-pak of 70% ethanol germinated fermented soybeans.

๋„ 9๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ๋ฐ 70% ์—ํƒ„์˜ฌ ์ถ”์ถœ๋ฌผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ์„ธํฌ์ƒ์กด์œจ์„ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 9 shows the cell viability of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.

๋„ 10์€ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ๋ฐ 70% ์—ํƒ„์˜ฌ ์ถ”์ถœ๋ฌผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ t-BHP์„ธํฌ์ƒ์กด์œจ์„ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 10 shows the growth rate of t- BHP cells of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.

๋„ 11์€ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ๋ฐ 70% ์—ํƒ„์˜ฌ ์ถ”์ถœ๋ฌผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ROS ์ธก์ • ๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 11 shows the results of ROS measurement of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.

๋„ 12๋Š” ๋™๋ฌผ๋ชจ๋ธ ํ˜ˆ์•กํ•™์ ๊ฒ€์‚ฌ AST(GOT), ALT(GPT) ์ธก์ •๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 12 shows the results of animal model hematology test AST (GOT), ALT (GPT).

๋„ 13์€ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ Liver์—์„œ์˜ MDA ์ธก์ •๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 13 shows the results of MDA measurement in Liver of the group administered t- BHP after the administration of germinated fermented soybean.

๋„ 14๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ Liver์—์„œ์˜ GST ์ธก์ •๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 14 shows the results of GST measurement in Liver of the group administered t- BHP after administration of germinated fermented soybeans.

๋„ 15๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ Liver์—์„œ์˜ GSH ์ธก์ •๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 15 shows the results of GSH measurement in Liver of the group administered t- BHP after administration of germinated fermented soybean.

๋„ 16์€ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ Liver์—์„œ์˜ SOD ์ธก์ •๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 16 shows the results of SOD measurement in Liver of the group administered t- BHP after the administration of germinated fermented soybean.

๋„ 17์€ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ Liver์—์„œ์˜ CAT ์ธก์ •๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ๊ฒƒ์ด๋‹ค.Figure 17 shows the results of CAT measurements in Liver of the group administered t- BHP after the administration of germinated fermented soybeans.

<์‹ค์‹œ์˜ˆ 1>; ์žฌ๋ฃŒ ๋ฐ ๋ฐฉ๋ฒ•<Example 1>; Materials and methods

1.1. ์‹คํ—˜์žฌ๋ฃŒ1.1. Experimental material

๋ณธ ์‹คํ—˜์— ์‚ฌ์šฉํ•œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์€ ํœด๋ฐ”์ด์˜ค๋กœ๋ถ€ํ„ฐ ์ œ๊ณต๋ฐ›์•˜๋‹ค. ์ œ์ฃผ ๋†์—…๊ธฐ์ˆ ์›(Jeju Agricultural Research and Extension Services)์œผ๋กœ๋ถ€ํ„ฐ ์–ป์€ ๋Œ€๋‘๋ฅผ ์„ ํ–‰์—ฐ๊ตฌ์— ๋”ฐ๋ผ ๋ฐœ์•„์‹œํ‚ค๊ณ  Aspergillus oryzae๋ฅผ ์ ‘์ข…ํ•˜์—ฌ ๋ฐœํšจ์‹œ์ผœ ์–ป๋Š”๋‹ค. ์ˆ˜์„ธ, ์ •์„  ๋ฐ ์ดˆ์ŒํŒŒ๋ฅผ ์ด์šฉํ•œ ์ถ”์ถœ ๊ณผ์ •์„ ๊ฑฐ์นœ ๋’ค, ๊ฑด์กฐ ํ•˜์—ฌ ์‹คํ—˜ ์‹œ๋ฃŒ๋กœ ์‚ฌ์šฉํ•˜์˜€๋‹ค.Germinated fermented soybean used in this experiment was provided by Hubio. Soybeans obtained from Jeju Agricultural Research and Extension Services are germinated and fermented by inoculation with Aspergillus oryzae according to previous studies. After the extraction process using water washing, choice and ultrasonic, it was dried and used as a test sample.

1.2. ์‹œ๋ฃŒ์˜ ์ œ์กฐ1.2. Preparation of Sample

๋Œ€๋‘๋Š” ์ˆ˜์„ธ, ์ •์„ , ๋ฐœ์•„ ๋ฐ Aspergillus oryzae๋ฅผ ์ด์šฉํ•œ ๋ฐœํšจ๊ณผ์ •์„ ๊ฑฐ์ณ ์—ฌ๊ณผํ•œ ๋’ค 60หšC์—์„œ ๊ฑด์กฐํ•˜์˜€๋‹ค. ๊ฑด์กฐ์‹œ๋ฃŒ 10 g์— 80% ์—ํƒ„์˜ฌ์„ 7 mL/g์œผ๋กœ ๊ฐ€ํ•˜์—ฌ ๋ถ„ํ•ดํ•œ ๋’ค, ์ดˆ์ŒํŒŒ๋ฅผ ์ด์šฉํ•œ ์—ด์ˆ˜์ถ”์ถœ์„ ํ•˜์˜€๋‹ค. ๊ทธ ํ›„, ์›์‹ฌ๋ถ„๋ฆฌ์„ ์ด์šฉํ•˜์—ฌ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์–ป์—ˆ๊ณ , ์ด๋ฅผ PTFE membrane์œผ๋กœ ์—ฌ๊ณผํ•œ ๋’ค, ๊ฑด์กฐํ•˜์—ฌ ์‹œ๋ฃŒ๋ฅผ ์–ป์—ˆ๋‹ค. ์œ„ ์‹œ๋ฃŒ๋ฅผ 3์ฐจ ์ฆ๋ฅ˜์ˆ˜์— ๋…น์—ฌ ๋™๋ฌผ์‹คํ—˜์— ์‚ฌ์šฉํ•˜์˜€๋‹คSoybeans wash, select, germinate and Aspergillus After the fermentation process using oryzae filtered and dried at 60หšC. 80% ethanol was added to 7 mL / g of 10 g of the dry sample, and the hydrothermal extract was extracted using ultrasonic waves. Thereafter, germinated fermented soybean extract was obtained by centrifugation, which was filtered through a PTFE membrane and dried to obtain a sample. The sample was dissolved in tertiary distilled water and used for animal experiments.

1.3. ์‹คํ—˜๋™๋ฌผ1.3. Laboratory animals

์‹คํ—˜๋™๋ฌผ์€ 8์ฃผ๋ น์˜ Sprague-Dawley๊ณ„ ์•”์ปท ํฐ์ฅ๋ฅผ ๋Œ€ํ•œ๋ฐ”์ด์˜ค๋งํฌ(๋Œ€์ „)๋กœ๋ถ€ํ„ฐ ๊ตฌ์ž…ํ•˜์—ฌ ๋ณธ ์‹คํ—˜์‹ค์—์„œ ๊ณ ํ˜•์‚ฌ๋ฃŒ(Purina)๋กœ ์‚ฌ์œกํ•˜์˜€๊ณ , ์‹คํ—˜ ์‹œ์ž‘ ์ „ 2์ฃผ์ผ ๋™์•ˆ ๋Œ€์กฐ๊ตฐ ์‹์ด๋กœ ์ ์‘์‹œํ‚จ ํ›„ ๋™๋ฌผ์˜ ์ฒด์ค‘์— ๋”ฐ๋ผ ๊ฐ ๊ตฐ์˜ ํ‰๊ท  ์ฒด์ค‘์„ 289.72ยฑ0.86 g์ด ๋˜๋„๋ก 7๋งˆ๋ฆฌ์”ฉ 6๊ตฐ[๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ(Negative control), ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ(sham), ์—์ŠคํŠธ๋กœ๊ฒ ํˆฌ์—ฌ๊ตฐ(OVX-estrogen), ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 0.5 g/kg ํˆฌ์—ฌ๊ตฐ(OVX-0.5), ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 1.0 g/kg ํˆฌ์—ฌ๊ตฐ(OVX-1.0) ๋ฐ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 2.0 g/kg ํˆฌ์—ฌ๊ตฐ(OVX-2.0)]์œผ๋กœ ๋‚˜๋ˆ„์–ด 6์ฃผ๊ฐ„ ์‹คํ—˜ํ•˜์˜€๋‹ค. ์‹คํ—˜๋™๋ฌผ์€ ์‚ฌ์œก์ผ€์ด์ง€ (20ร—26ร—13 cm)๋ฅผ ์ด์šฉํ•ด ์‹คํ—˜์‹ค ์˜จ๋„ 22-24โ„ƒ, ์Šต๋„ 60ยฑ5%๊ฐ€ ์œ ์ง€๋˜๋ฉฐ ๋ฐค๋‚ฎ ์ฃผ๊ธฐ(12 ์‹œ๊ฐ„ light/ 12 ์‹œ๊ฐ„ dark)๊ฐ€ ์ž๋™ ์กฐ์ ˆ ์žฅ์น˜์— ์˜ํ•ด ์กฐ์ ˆ๋˜๋Š” ๊ณ ๋ ค๋Œ€ํ•™๊ต ๋™๋ฌผ์‹ค์—์„œ ์‚ฌ์œกํ•˜๋ฉฐ ๊ณ ๋ ค๋Œ€ํ•™๊ต ๋™๋ฌผ์‹คํ—˜ ์œค๋ฆฌ์œ„์›ํšŒ ์Šน์ธ์„ ๋ฐ›์€ ํ›„ '์‹คํ—˜๋™๋ฌผ ๊ด€๋ฆฌ ๋ฐ ์ด์šฉ์— ๊ด€ํ•œ ์ง€์นจ (Guide for the care and use of Laboratory Animals, NRC)'์— ๋งž์ถ”์–ด ๊ด€๋ฆฌํ•˜๋ฉด์„œ ์‹ค์‹œํ•˜์˜€๋‹ค. ๋ฌผ๊ณผ ์‹์ด๋Š” ์ž์œ  ๊ณต๊ธ‰ํ•˜์˜€๊ณ , ์‹คํ—˜ ์‹œ๋ฃŒ๋Š” 3์ฐจ ์ฆ๋ฅ˜์ˆ˜์— ์šฉํ•ดํ•˜์—ฌ ๋งค์ผ 3 ml์”ฉ ๊ฒฝ๊ตฌ ํˆฌ์—ฌ ํ•˜์˜€๊ณ , ๋Œ€์กฐ๊ตฐ(Negative-control, sham)์€ ๋™์ผ์šฉ๋Ÿ‰์˜ 3์ฐจ ์ฆ๋ฅ˜์ˆ˜๋ฅผ ๋งค์ผ ๊ฒฝ๊ตฌ ํˆฌ์—ฌํ•˜์˜€๋‹ค.The experimental animals were purchased from Korea Biolink (Daejeon), 8-week-old Sprague-Dawley female rats, and were bred as solid feed (Purina) in the laboratory. Therefore, the average body weight of each group was 289.72 ยฑ 0.86 g, and 7 animals of 6 groups (Negative control, Non-ovarian control group (sham), Estrogen group (OVX-estrogen), Germinated fermented soybean extract 0.5 g / kg Administered group (OVX-0.5), germinated fermented soybean extract 1.0 g / kg administered group (OVX-1.0) and germinated fermented soybean extract 2.0 g / kg administered group (OVX-2.0)]. The experimental animals were kept in a laboratory temperature of 22-24 ยฐ C, humidity 60 ยฑ 5% using a breeding cage (20 ร— 26 ร— 13 cm), and the day and night cycle (12 hours light / 12 hours dark) was controlled by an automatic control device. It was raised in the university animal room and approved by Korea University's Animal Experiment Ethics Committee, and managed according to the 'Guide for the care and use of Laboratory Animals' (NRC). Water and diet were freely supplied, and the experimental samples were dissolved in tertiary distilled water and orally administered 3 ml daily, and the control group (Negative-control, sham) was orally administered daily with the same dose of tertiary distilled water.

1.4. ๋‚œ์†Œ ์ ˆ์ œ ์‹œ์ˆ 1.4. Ovarian excision procedure

2์ฃผ์ผ ๋™์•ˆ ์ฃผ์œ„ํ™˜๊ฒฝ์— ์ ์‘์‹œ์ผœ ์ฒด์ค‘์— ๋”ฐ๋ผ ๋‚œ๊ดด๋ฒ•(Randomized Complete Block Design)์— ์˜ํ•ด ๊ตฐ์„ ๋‚˜๋ˆ„์–ด ๋‚œ์†Œ์ ˆ์ œ ์‹œ์ˆ ์„ ํ•˜์˜€๋‹ค. ์ˆ˜์ˆ ์€ isoflurane์œผ๋กœ ๋งˆ์ทจ ํ›„ ์‹ฌ๋งˆ์ทจ๊ธฐ์— ์ด๋ฅด๋ฉด ๋“ฑ์ชฝ ์˜†๊ตฌ๋ฆฌ๋ฅผ ์ ˆ๊ฐœํ•˜์—ฌ ๋‚œ์†Œ๋ฅผ ์ œ๊ฑฐํ•˜๊ณ  ์ ˆ๊ฐœ๋ถ€์œ„๋ฅผ ๋ด‰ํ•ฉํ•˜์˜€๋‹ค. ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ(Sham)์˜ ๊ฒฝ์šฐ, ๊ฐ™์€ ๋ฐฉ๋ฒ•์œผ๋กœ ๋‚œ์†Œ ์˜†์˜ ์ง€๋ฐฉ์„ ์ œ๊ฑฐํ•˜๊ณ  ์ ˆ๊ฐœ๋ถ€์œ„๋ฅผ ๋ด‰ํ•ฉํ•˜์˜€๋‹ค. ์‹œ๋ฃŒ๋Š” ์ˆ˜์ˆ  ํ›„ 2์ฃผ๊ฐ„์˜ ํšŒ๋ณต๊ธฐ๋ฅผ ๊ฐ€์ง„ ํ›„์— ๊ฒฝ๊ตฌ ํˆฌ์—ฌํ•˜์˜€๋‹ค.For 2 weeks, ovarian resection was performed by dividing the group by randomized complete block design according to body weight. Surgery was performed with isoflurane and after anesthesia, the dorsal flank was incised to remove the ovary and the incision was closed. In the case of non-ovarian resection control (Sham), fat was removed from the ovary in the same way and the incision was closed. Samples were administered orally after having a recovery period of 2 weeks after surgery.

1.5. ํ˜ˆ์ฒญ ์ง€์งˆ ๋†๋„ ๋ฐ ํšจ์†Œ ํ™œ์„ฑ ๋ถ„์„1.5. Serum Lipid Concentration and Enzyme Activity Analysis

๋ถ„๋ฆฌํ•œ ํ˜ˆ์ฒญ์˜ ์ค‘์„ฑ์ง€๋ฐฉ(TG), ์ด ์ฝœ๋ ˆ์Šคํ…Œ๋กค(TCHO), ๊ณ ๋ฐ€๋„์ง€๋‹จ๋ฐฑ ์ฝœ๋ ˆ์Šคํ…Œ๋กค(HDL), ์ €๋ฐ€๋„์ง€๋‹จ๋ฐฑ ์ฝœ๋ ˆ์Šคํ…Œ๋กค(LDL), ํฌ๋„๋‹น(GLU)์€ FUJI DRI-CHEM 3500๋ฅผ ์ด์šฉํ•˜์—ฌ ์ธก์ •ํ•˜์˜€๋‹ค. Glutamate oxaloacetate transaminase(GOT), Glutamate pyruvate transmaminase (GPT) ํ™œ์„ฑ ๋˜ํ•œ ๊ฐ™์€ ๋ฐฉ๋ฒ•์œผ๋กœ ์ธก์ •ํ•˜์˜€๋‹ค.The triglyceride (TG), total cholesterol (TCHO), high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL) and glucose (GLU) of the isolated serum were measured using FUJI DRI-CHEM 3500. Glutamate oxaloacetate transaminase (GOT) and Glutamate pyruvate transmaminase (GPT) activities were also measured in the same way.

1.6. ํ˜ธ๋ฅด๋ชฌ ์ธก์ •1.6. Hormone measurement

ํ˜ˆ์žฅ ๋‚ด estradiol, testosterone์˜ ์–‘์€ ELISA๋ฒ•์„ ์ด์šฉํ•˜์—ฌ ๊ฐ๊ฐ rat estradiol Elisa kit(Enzo life science, NY, USA)๊ณผ rat testosterone Elisa kit(Enzo life science, NY, USA) ๋กœ ์ธก์ •ํ•˜์˜€๋‹ค.Plasma levels of estradiol and testosterone were measured by the rat estradiol Elisa kit (Enzo life science, NY, USA) and rat testosterone Elisa kit (Enzo life science, NY, USA) by ELISA.

1.7. ๊ณจ๋Œ€์‚ฌ ์ง€ํ‘œ ์ธก์ • 1.7. Bone metabolism indicator measurement

๊ณจํ˜•์„ฑ ์ง€ํ‘œ๋กœ ํ˜ˆ์žฅ ์นผ์Š˜๊ณผ ์ธ์˜ ๋†๋„, ํ˜ˆ์žฅ alkaline phosphatase(ALP)์˜ ๋†๋„๋Š” FUJI DRI-CHEM 3500๋กœ ์ธก์ •ํ•˜์˜€์œผ๋ฉฐ BONE-alkaline phosphatase(BONE-ALP)์€ ์„ ํ–‰ ์—ฐ๊ตฌ์— ๋”ฐ๋ผ ์ธก์ •ํ•˜์˜€๋‹ค. ๋Œ€ํ‡ด๊ณจ์„ ๊ฑด์กฐ, ์ œ์ง€๋ฐฉ ๊ณผ์ •์„ ๊ฑฐ์ณ ํšŒํ™”ํ•˜์˜€๊ณ , ๊ทธ ํ›„ ๋Œ€ํ‡ด๊ณจ์˜ ๊ณจ์—ผ๋Ÿ‰์„ ์ธก์ •ํ•˜์˜€๋‹ค.Plasma calcium and phosphorus concentrations and alkaline alkaline phosphatase (ALP) concentrations were measured by FUJI DRI-CHEM 3500 and BONE-alkaline phosphatase (BONE-ALP) as bone markers. The femur was dried and crushed through the fat removal process, and then the bone salt in the femur was measured.

1.8. ๊ณจ์กฐ์ง ํ˜•ํƒœ ์ธก์ •: Micro-CT ์ดฌ์˜1.8. Bone Morphology Measurement: Micro-CT

ํฌ์ƒ ์ฅ์˜ ๊ฒฝ๊ณจ ์ค‘ ์„ฑ์žฅํŒ์œผ๋กœ๋ถ€ํ„ฐ 1mm ๋–จ์–ด์ง„ ๋ถ€์œ„๋ฅผ 2mm ๋†’์ด ๋ถ€๋ถ„์„ Micro-CT(Skyscan1072, SKYSCAN, Antwerpen, Belgium)๋ฅผ ์ด์šฉํ•˜์—ฌ ์ดฌ์˜ํ•˜์˜€๋‹ค. ์ดฌ์˜ ๊ด€์ „์••์€ 50 kVp, ๊ด€์ „๋ฅ˜๋Š” 500 ฮผA, 0.5 mm ์•Œ๋ฃจ๋ฏธ๋Š„ ์—ฌ๊ณผ๋ฅผ ์ด์šฉํ•˜์˜€๊ณ , ์ดฌ์˜์‹œ๊ฐ„์€ 210 ms์ด์—ˆ์œผ๋ฉฐ, ํ™”์†Œ ํฌ๊ธฐ(pixel size)๋Š” 18 ฮผm์˜€๋‹ค. ๊ฐœ๊ฐœ์˜ ์˜์ƒ์—์„œ ๊ณจ์†Œ์ฃผ์™€ ๊ณจ์ˆ˜๊ฐ•์„ ๋ถ„๋ฆฌํ•˜๊ธฐ ์œ„ํ•ด ์—ญ์น˜๊ฐ’(threshold value)์„ 58๋กœ ๊ณ ์ •ํ•˜์˜€์œผ๋ฉฐ 3์ฐจ์› ์˜์ƒ์„ ์žฌ๊ตฌ์„ฑํ•˜์˜€๋‹ค. Micro-CT๋กœ ์ดฌ์˜๋œ ์˜์ƒ๋“ค๋กœ๋ถ€ํ„ฐ SkyscanTMCT-analyzersoftware๊ฐ€ ์‚ฌ์šฉ๋˜์–ด ๊ณจ ๋ฏธ์„ธ ๊ตฌ์กฐ ์ง€ํ‘œ๋“ค์ด ๋ถ„์„๋˜์—ˆ๋‹ค. ์ด์šฉ ๋ถ„์„๋œ ์ง€ํ‘œ๋“ค์€ Percent bone volume (BV/TV (%)), trabecular number (Tb.N ()), trabecular thickness (Tb.Th (ฮผm)), trabecular separation (Tb.Sp (ฮผm)), trabecular pattern factor (Tb.pf ()), and structure model index (SMI) ์ด์—ˆ๋‹ค.A 2 mm high part of the tibia of the sacrificial rats was photographed using Micro-CT (Skyscan1072, SKYSCAN, Antwerpen, Belgium). The imaging tube voltage was 50 kVp, the tube current was 500 ฮผA, 0.5 mm aluminum filtration, the imaging time was 210 ms, and the pixel size was 18 ฮผm. The threshold value was fixed at 58 to separate the bone column and bone marrow from the individual images, and the 3D image was reconstructed. Bone microstructure indicators were analyzed using Skyscan โ„ข CT-analyzersoftware from images taken with Micro-CT. The indicators analyzed were Percent bone volume (BV / TV (%)), trabecular number (Tb.N ()), trabecular thickness (Tb.Th (ฮผm)), trabecular separation (Tb.Sp (ฮผm)), trabecular pattern factor (Tb.pf ()), and structure model index (SMI).

1.9. ํ†ต๊ณ„์ฒ˜๋ฆฌ1.9. Statistical processing

์—ฐ๊ตฌ๊ฒฐ๊ณผ ์–ป์–ด์ง„ ์ž๋ฃŒ๋ฅผ SPSS (statistical package for social science, version 12.0) ํ†ต๊ณ„ ํ”„๋กœ๊ทธ๋žจ์„ ์‚ฌ์šฉํ•˜์—ฌ ์ฒ˜๋ฆฌํ•˜์˜€๋‹ค. ์ง‘๋‹จ๊ฐ„์˜ ์œ ์˜์ ์ธ ์ฐจ์ด๋ฅผ ์•Œ์•„๋ณด๊ธฐ ์œ„ํ•ด Negative-control ๊ตฐ์— ๋Œ€ํ•œ T-test๋ฅผ ์‹ค์‹œํ•˜์˜€๋‹ค. ์ธก์ •๊ฐ’์€ ํ‰๊ท  ํ‘œ์ค€์˜ค์ฐจ๋กœ ํ‘œ๊ธฐํ•˜์˜€์œผ๋ฉฐ, p<0.05, p<0.01, p<0.001 ์€ ๊ฐ๊ฐ *, **, *** ๋กœ ํ‘œ๊ธฐ๋˜์—ˆ๋‹ค.The data obtained were processed using the statistical package for social science (version 12.0) statistical program. T-test was performed on the negative-control group to identify the significant differences among the groups. The measured values are expressed as average standard error, and p <0.05, p <0.01, and p <0.001 are indicated by *, **, and *** , respectively.

<์‹ค์‹œ์˜ˆ 1> ์‹คํ—˜๊ฒฐ๊ณผExample 1 Experimental Results

2.1. ์ฒด์ค‘ ๋ณ€ํ™” ๋ฐ ์‹์ด์„ญ์ทจ๋Ÿ‰2.1. Weight change and dietary intake

6 ์ฃผ๊ฐ„ ์‹คํ—˜์„ ์ง„ํ–‰ํ•˜๋Š” ๋™์•ˆ, ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ๋‚œ์†Œ์ ˆ์ œ์ˆ ๋กœ ๊ฐฑ๋…„๊ธฐ๋ฅผ ์œ ๋„ํ•œ ๋ชจ๋“  ๊ตฐ์—์„œ ์ฒด์ค‘์ด ์ฆ๊ฐ€ํ•˜๋Š” ๊ฒฝํ–ฅ์„ ํ™•์ธํ–ˆ๋‹ค(๋„ 1). ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์˜ ์ฒด์ค‘ ์ฆ๊ฐ€๋Ÿ‰์€ 20.4 ยฑ 3.3 g ์ด๊ณ , ์ด์— ๋น„ํ•ด ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ 0.5 g/kg, 1.0 g/kg, 2.0 g/kg๋กœ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ ์ฒด์ค‘ ์ฆ๊ฐ€๋Ÿ‰์€ ๊ฐ๊ฐ 39.8 ยฑ 11.1 g, 42.2 ยฑ 4.5 g, 34.0 ยฑ 5.6 g, 36.8 ยฑ 4.3 g ์ž„์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค. ์—์ŠคํŠธ๋กœ๊ฒ์„ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ ์ฒด์ค‘ ์ฆ๊ฐ€๋Ÿ‰์€ 12.1 ยฑ 8.7g ์œผ๋กœ, ๋น„๊ต์  ๊ฐ์†Œํ•˜๋Š” ๊ฒฝํ–ฅ์„ ๋ณด์˜€์œผ๋‚˜, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ํˆฌ์—ฌ ๊ตฐ์—์„œ๋Š” ์œ ์˜์ ์ธ ์ฐจ์ด๊ฐ€ ๋ณด์ด์ง€ ์•Š์•˜๋‹ค. ์‹์ด์„ญ์ทจ๋Ÿ‰ ๋ณ€ํ™”์—์„œ๋„ ์œ ์˜์ ์ธ ์ฐจ์ด๋ฅผ ๋ณด์ด์ง€ ์•Š์•˜๋‹ค. ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ, ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ, ์—์Šคํ† ๋กœ๊ฒ ํˆฌ์—ฌ๊ตฐ์€ ๊ฐ Neg.C, Sham, Estradiol๋กœ ํ‘œ๊ธฐํ•˜์˜€๊ณ , ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 0.5g/kg, 1.0g/kg, 2.0g/kg ํˆฌ์—ฌ๊ตฐ์€ ๊ฐ๊ฐ 0.5, 1.0, 2.0์œผ๋กœ ํ‘œ๊ธฐํ•˜์˜€๋‹ค. ์ˆ˜์น˜๋Š” ํ‰๊ท  ยฑ ํ‘œ์ค€์˜ค์ฐจ๋กœ ํ‘œ๊ธฐํ•˜์˜€๋‹ค.During the six-week experiment, body weight gain was observed in all groups induced by menopause by ovarian resection compared to the non-ovarian resection control (FIG. 1). The weight gain of the non-ovarian control group was 20.4 ยฑ 3.3 g, whereas the weight gain of the ovarian control group and the germinated fermented soybean at 0.5 g / kg, 1.0 g / kg and 2.0 g / kg were 39.8 ยฑ 11.1, respectively. g, 42.2 ยฑ 4.5 g, 34.0 ยฑ 5.6 g, 36.8 ยฑ 4.3 g was confirmed. The weight gain of the estrogen-treated group was 12.1 ยฑ 8.7g, which tended to decrease relatively, but there was no significant difference in the germinated fermented soybean group. There was no significant difference in dietary intake changes. The ovarian resection control group, non-ovarian resection control group, and the estrogen-administered group were labeled as Neg.C, Sham, and Estradiol, and the germinated fermented soybean 0.5g / kg, 1.0g / kg, and 2.0g / kg groups were 0.5, 1.0, and 2.0, respectively. Notation. The figures are expressed as mean ยฑ standard error.

2.2. ํ˜ˆ์ฒญ ์ง€์งˆ ๋†๋„ ๋ฐ ํšจ์†Œ ํ™œ์„ฑ ๋ถ„์„2.2. Serum Lipid Concentration and Enzyme Activity Analysis

์—ฌ์„ฑ ๊ฐฑ๋…„๊ธฐ์™€ ๋งˆ์ฐฌ๊ฐ€์ง€๋กœ, ๋‚œ์†Œ์ ˆ์ œ๋ฅผ ์‹ค์‹œํ•œ ๊ทธ๋ฃน์—์„œ๋Š” ์—์ŠคํŠธ๋กœ๊ฒ ๋ถ„๋น„๊ฐ€ ๊ธ‰๊ฒฉํ•˜๊ฒŒ ๊ฐ์†Œํ•œ๋‹ค. ์ด๋•Œ ์‹ ์ฒด๋Š” ๊ฐ์†Œํ•œ ์—์ŠคํŠธ๋กœ๊ฒ ๋ถ„๋น„๋ฅผ ๋ณด๊ฐ•ํ•˜๊ธฐ ์œ„ํ•˜์—ฌ ์—์ŠคํŠธ๋กœ๊ฒ์„ ์ƒ์‚ฐํ•  ์ˆ˜ ์žˆ๋Š” ์ง€๋ฐฉ ์„ธํฌ๋ฅผ ๋งŽ์ด ์ถ•์ฒ™ํ•˜๊ฒŒ ๋˜๊ณ  ์ด๋กœ ์ธํ•ด ๊ฐฑ๋…„๊ธฐ ๋น„๋งŒ์ด ์œ ๋„๋œ๋‹ค. ๊ฐฑ๋…„๊ธฐ ๋น„๋งŒ์˜ ์ง€ํ‘œ๋กœ ํ˜ˆ์•ก ๋‚ด์˜ ์ง€์งˆ, ์ฝœ๋ ˆ์Šคํ…Œ๋กค, ํฌ๋„๋‹น ๋†๋„๋ฅผ ํ™•์ธํ•˜์˜€๊ณ  ๊ทธ ๊ฒฐ๊ณผ๋Š” ๋„ 2์™€ ๊ฐ™๋‹ค. As in female menopause, estrogen secretion decreases rapidly in the ovarian resection group. At this time, the body accumulates a lot of fat cells capable of producing estrogen in order to enhance the reduced estrogen secretion, which leads to menopausal obesity. Lipid, cholesterol, and glucose concentrations in blood were identified as indicators of menopausal obesity, and the results are shown in FIG. 2.

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ 0.5 g/kg, 1.0 g/kg, 2.0 g/kg๋กœ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ ์ค‘์„ฑ ์ง€๋ฐฉ ์ˆ˜์น˜๋Š” ๊ฐ๊ฐ 62.5 ยฑ 21.3 mg/dl, 59.0 ยฑ 8.5 mg/dl, 55.8 ยฑ 8.8 mg/dl์œผ๋กœ, ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ 66.8 ยฑ 12.6 mg/dl ๊ณผ ์œ ์‚ฌํ•œ ์ˆ˜์น˜๋กœ ๊ฐ์†Œํ•˜๋Š” ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค. Neutral fat levels of the groups administered with 0.5 g / kg, 1.0 g / kg and 2.0 g / kg of germinated fermented soybean were 62.5 ยฑ 21.3 mg / dl, 59.0 ยฑ 8.5 mg / dl and 55.8 ยฑ 8.8 mg / dl, respectively. The decrease was similar to that of the non-ovarian ablation control 66.8 ยฑ 12.6 mg / dl.

์ด ์ฝœ๋ ˆ์Šคํ…Œ๋กค์˜ ๊ฒฝ์šฐ, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 1.0 g/kg, 2.0 g/kg๋กœ ํˆฌ์—ฌํ•œ ๊ฒฝ์šฐ ๊ฐ๊ฐ 139.5 ยฑ 9.6 mg/dl, 136.7 ยฑ 6.5 mg/dl์œผ๋กœ ์ˆ˜์น˜๊ฐ€ ์ฆ๊ฐ€ํ•œ ๊ฒƒ์ด ํ™•์ธ๋˜์—ˆ๋‹ค. ๊ฒฐ๊ณผ์ ์œผ๋กœ cholesterol pool์ด ์ปค์ ธ ๊ณ ๋ฐ€๋„์ง€๋‹จ๋ฐฑ ์ฝœ๋ ˆ์Šคํ…Œ๋กค๊ณผ ์ €๋ฐ€๋„์ง€๋‹จ๋ฐฑ ์ฝœ๋ ˆ์Šคํ…Œ๋กค์—์„œ๋„ ์œ ์‚ฌํ•œ ํšจ๊ณผ๊ฐ€ ๋‚˜ํƒ€๋‚œ ๊ฒƒ์œผ๋กœ ๋ณด์ธ๋‹ค. In the case of total cholesterol, the values increased to 139.5 ยฑ 9.6 mg / dl and 136.7 ยฑ 6.5 mg / dl, respectively, when administered at 1.0 g / kg and 2.0 g / kg of germinated fermented soybean. As a result, the cholesterol pool was enlarged, and similar effects were observed in high density lipoprotein cholesterol and low density lipoprotein cholesterol.

๋„ 2๋ฅผ ์‚ดํŽด๋ณด๋ฉด, ํ˜ˆ์ค‘ ํฌ๋„๋‹น์˜ ๋†๋„ ๋˜ํ•œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ๋ชจ๋“  ๊ตฐ์—์„œ 98.3 ยฑ 16.1 mg/dl, 123.3 ยฑ 14.4 mg/dl, 112.8 ยฑ 6.6 mg/dl์œผ๋กœ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์˜ ํ˜ˆ์ค‘ ํฌ๋„๋‹น ๋†๋„์ธ 162.7 ยฑ18.6 mg/dl ์— ๋น„ํ•ด ๊ฐ์†Œํ•˜๋Š” ๊ฒฝํ–ฅ์ด ๋‚˜ํƒ€๋‚˜๋Š” ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค.Referring to Figure 2, the blood glucose concentration was also 98.3 ยฑ 16.1 mg / dl, 123.3 ยฑ 14.4 mg / dl, 112.8 ยฑ 6.6 mg / dl in all groups administered with germinated fermented soybeans, the blood glucose concentration of 162.7 ยฑ It was confirmed that the tendency to decrease compared to 18.6 mg / dl.

ํ˜ˆ์•ก ๋‚ด ์ค‘์„ฑ์ง€๋ฐฉ์˜ ๋†๋„์™€ ํฌ๋„๋‹น์˜ ๋†๋„๊ฐ€ ๊ฐ์†Œํ•˜๋Š” ๊ฒฝํ–ฅ์„ ๊ฐ–๋Š” ๊ฒƒ์œผ๋กœ ๋ณด์•„, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ํˆฌ์—ฌ๊ฐ€ ๊ฐฑ๋…„๊ธฐ ๋น„๋งŒ ๊ฐœ์„ ์— ๋„์›€์„ ์ค„ ๊ฒƒ์œผ๋กœ ์˜ˆ์ƒ๋œ๋‹ค.Given that the concentration of triglycerides and glucose in the blood tend to decrease, the administration of germinated fermented soybean is expected to help improve menopausal obesity.

GOT, GPT๋Š” ๊ฐ„์—์„œ ๋งŒ๋“ค์–ด์ง€๋Š” ์ •์ƒ ํšจ์†Œ์ด๋‹ค. ๊ฐ„์ด ์†์ƒ ๋ฐ›์„ ๊ฒฝ์šฐ ๊ฐ„์„ธํฌ๊ฐ€ ๊นจ์ง€๋ฉด์„œ ์ด ํšจ์†Œ๋“ค์ด ํ˜ˆ์•ก์œผ๋กœ ์œ ๋ฆฌ๋˜์–ด ์ •์ƒ๋ณด๋‹ค ๋งŽ์ด ์ฆ๊ฐ€ํ•˜๊ธฐ ๋•Œ๋ฌธ์— ์ด๋“ค์€ ๋Œ€ํ‘œ์ ์ธ ๊ฐ„ ๋…์„ฑ์˜ ์ง€ํ‘œ๋กœ ์‚ฌ์šฉ๋œ๋‹ค. ํ•˜๊ธฐ ํ‘œ 1์„ ์‚ดํŽด๋ณด๋ฉด ํ˜ˆ์ฒญ GOT, GPT ํ™œ์„ฑ ๋ถ„์„์€ ๊ทธ๋ฃน ๋ณ„๋กœ ์œ ์˜์ ์ธ ์ฐจ์ด๋ฅผ ๊ฐ–์ง€ ์•Š๋Š”๋‹ค. ๋”ฐ๋ผ์„œ ์‹œ๋ฃŒ์˜ ์ฒ˜๋ฆฌ๊ฐ€ ๊ฐ„ ๋…์„ฑ์„ ๋‚˜ํƒ€๋‚ด์ง€ ์•Š๋Š” ๊ฒƒ์„ ์•Œ ์ˆ˜ ์žˆ๋‹ค. GOT and GPT are normal enzymes made in the liver. When the liver is damaged, these cells are broken down and the enzymes are released into the blood, increasing more than normal, so they are used as representative indicators of liver toxicity. Looking at Table 1 below, serum GOT, GPT activity analysis does not have a significant difference between groups. Therefore, it can be seen that the treatment of the sample does not show liver toxicity.

ToxicityToxicity ShamSham Neg.CNeg.C EstradiolEstradiol 0.50.5 1.01.0 2.02.0 GOT(U/L)GOT (U / L) 92.00ยฑ10.4292.00 ยฑ 10.42 76.57ยฑ7.2876.57 ยฑ 7.28 63.71ยฑ5.8963.71 ยฑ 5.89 62.71ยฑ6.3562.71 ยฑ 6.35 74.00ยฑ3.9174.00 ยฑ 3.91 69.14ยฑ5.1169.14 ยฑ 5.11 GPT(U/L)GPT (U / L) 28.43ยฑ3.9028.43 ยฑ 3.90 23.86ยฑ1.5823.86 ยฑ 1.58 31.00ยฑ3.7231.00 ยฑ 3.72 22.00ยฑ6.2522.00 ยฑ 6.25 23.86ยฑ0.5923.86 ยฑ 0.59 26.43ยฑ5.4226.43 ยฑ 5.42

2.3. ํ˜ธ๋ฅด๋ชฌ ์ธก์ •2.3. Hormone measurement

๋‹ค์–‘ํ•œ ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์˜ ์›์ธ์€ ๋‚œ์†Œ์˜ ๋…ธํ™”๋กœ ์ธํ•œ ์—์ŠคํŠธ๋กœ๊ฒ ์ƒ์‚ฐ๊ณผ ๋ถ„๋น„์˜ ๊ฐ์†Œ์ด๋‹ค. ๋”ฐ๋ผ์„œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ํˆฌ์—ฌ์˜ ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์„ ์™„ํ™” ํšจ๊ณผ๋ฅผ ๋ณด๊ธฐ ์œ„ํ•ด์„œ๋Š” ํ˜ˆ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„ ์ฆ๊ฐ€๋ฅผ ํ™•์ธํ•˜๋Š” ๊ฒƒ์ด ์ค‘์š”ํ•˜๋‹ค. The cause of various menopausal symptoms is a decrease in estrogen production and secretion due to aging of the ovary. Therefore, it is important to confirm the increase of blood estrogen concentration in order to alleviate the menopausal symptoms of the administration of germinated fermented soybeans.

2.3.1. ์—์ŠคํŠธ๋กœ๊ฒ2.3.1. Estrogen

๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์˜ ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„ 1020. ยฑ 63.6 pg/ml์™€ ๋น„๊ตํ•˜์—ฌ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์˜ ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„๊ฐ€ 500.3 ยฑ 27.8 pg/ml์ธ ๊ฒƒ์œผ๋กœ ๋ณด์•„, ๋‚œ์†Œ์ ˆ์ œ๋ฅผ ํ†ตํ•ด ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ (p < 0.001) ๊ฐ์†Œํ•œ ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ๋‹ค. ์—์ŠคํŠธ๋กœ๊ฒ ํˆฌ์—ฌ๊ตฐ์˜ ๊ฒฝ์šฐ, ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ๊ณผ ์œ ์‚ฌํ•œ ์ˆ˜์น˜์ธ 1134.1 ยฑ 205.8 pg/ml๋กœ ํ˜ˆ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„๊ฐ€ ์ฆ๊ฐ€ํ•œ ๊ฒƒ์„ ํ™•์ธํ•˜์˜€๋‹ค(๋„ 3). ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ์„ธ ๊ตฐ ๋ชจ๋‘ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ๊ณผ ๋น„๊ตํ•˜์—ฌ ๊ฐ๊ฐ 1577.1 ยฑ 341.6 pg/ml, 2358.6 ยฑ 191.8 pg/ml, 938.5 ยฑ 51.2 pg/ml ์œ ์˜์ ์œผ๋กœ ํ˜ˆ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„๊ฐ€ ์ฆ๊ฐ€ํ•˜์˜€๋‹ค(๋„ 3). ํ˜ˆ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ ๋†๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ•˜๋Š” ๊ฒƒ์„ ํ†ตํ•ด ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ์ฟ ๋ฉ”์Šคํ…Œ๋กค์„ ํ•จ์œ ํ•œ ์ถ”์ถœ๋ฌผ์ด ์—์ŠคํŠธ๋กœ๊ฒ์˜ ์—ญํ• ์„ ๋Œ€์ฒดํ•  ์ˆ˜ ์žˆ๊ณ  ์ด๋กœ ์ธํ•ด ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์ด ์™„ํ™”๋  ๊ฒƒ์„ ๊ธฐ๋Œ€ํ•  ์ˆ˜ ์žˆ๋‹ค.The estrogen concentration of the ovarian resection control group was 500.3 ยฑ 27.8 pg / ml compared to the estrogen concentration of 1020. ยฑ 63.6 pg / ml of the non-ovarian resection group, resulting in a significant (p <0.001) decrease in estrogen concentration. You can see that. In the estrogen-administered group, it was confirmed that blood estrogen concentration was increased to 1134.1 ยฑ 205.8 pg / ml, which is similar to that of the non-ovarian ablation control group (FIG. 3). All three groups that received germinated fermented soybeans significantly increased blood estrogen levels in 1577.1 ยฑ 341.6 pg / ml, 2358.6 ยฑ 191.8 pg / ml, and 938.5 ยฑ 51.2 pg / ml, respectively, as compared to the ovarian ablation control (FIG. 3). Significantly increased blood estrogen levels can be expected to reduce the role of estrogens by extracts containing cousterol from germinated fermented soybeans, thereby alleviating menopausal symptoms.

2.3.2. ํ…Œ์Šคํ† ์Šคํ…Œ๋ก 2.3.2. Testosterone

ํ…Œ์Šคํ† ์Šคํ…Œ๋ก ์€ ๋Œ€ํ‘œ์ ์ธ ๋™ํ™” ํ˜ธ๋ฅด๋ชฌ (Anabolic hormone)์œผ๋กœ, ๊ทผ์œก๋Œ€์‚ฌ์™€ ์ง€๋ฐฉ๋Œ€์‚ฌ์— ๊ด€์—ฌํ•œ๋‹ค. ์ด ํ˜ธ๋ฅด๋ชฌ์˜ ์ˆ˜์น˜๊ฐ€ ๋‚ฎ์•„์งˆ ๊ฒฝ์šฐ, ์ฒด๋‚ด ๊ทผ์œก๋Ÿ‰์ด ๊ฐ์†Œํ•˜์—ฌ ๊ธฐ์ดˆ๋Œ€์‚ฌ๋Ÿ‰์ด ๋‚ฎ์•„์ง€๊ณ  ๋น„๋งŒ์„ ์‰ฝ๊ฒŒ ์•ผ ๊ธฐํ•˜๊ฒŒ ๋œ๋‹ค. ๋˜ํ•œ ๊ฐฑ๋…„๊ธฐ์˜ ๋Œ€ํ‘œ์ ์ธ ์ฆ์ƒ ์ค‘ ํ•˜๋‚˜์ธ ํ”ผ๋ถ€ ๊ฐ์งˆํ™”๋ฅผ ์œ ๋„ํ•˜๊ฒŒ ๋œ๋‹ค. ๋„ 3์„ ์‚ดํŽด๋ณด๋ฉด, ๋‚œ์†Œ์ ˆ์ œ๋ฅผ ์‹ค์‹œํ•œ ๋ชจ๋“  ๊ตฐ์—์„œ ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ํ˜ˆ์ค‘ ํ…Œ์Šคํ† ์Šคํ…Œ๋ก ์˜ ๋†๋„๊ฐ€ ๊ฐ์†Œํ•˜์˜€๋‹ค. ์ด ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ ํˆฌ์—ฌ๊ตฐ๊ณผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 1.0 g/kg๋กœ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ๋Š” ๊ฐ๊ฐ 2797.8 ยฑ 517.3 pg/ml, 1969.8 ยฑ 164.2 pg/ml์˜ ํ…Œ์ŠคํŠธ์Šคํ† ์Šคํ…Œ๋ก  ๋†๋„๋ฅผ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๊ณ , ์ด ๊ทธ๋ฃน๋“ค์˜ ํ…Œ์Šคํ† ์Šคํ…Œ๋ก  ๋†๋„๋Š” ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ ๊ตฐ์— ๋น„ํ•ด ์œ ์˜์ ์œผ๋กœ (p < 0.05) ์ฆ๊ฐ€ํ•œ ๊ฒฐ๊ณผ์ž„์„ ํ™•์ธํ•˜์˜€๋‹ค.Testosterone is a representative anabolic hormone that is involved in muscle and fat metabolism. Lower levels of these hormones reduce muscle mass in the body, which leads to lower basal metabolism and easier obesity. It also induces skin keratinization, one of the typical symptoms of menopause. Referring to FIG. 3, the concentration of testosterone in blood was decreased in all the groups that underwent ovarian ablation compared to the non-ovarian ablation control group. Among them, the testosterone concentrations of 2797.8 ยฑ 517.3 pg / ml and 1969.8 ยฑ 164.2 pg / ml were found in the estrogen-administered group and 1.0 g / kg of germinated fermented soybean, respectively. It was confirmed that the results were significantly increased (p <0.05) compared to the control group.

2.4. ๊ณจ๋Œ€์‚ฌ ์ง€ํ‘œ ์ธก์ • 2.4. Bone metabolism indicator measurement

๋„ 4๋ฅผ ๋ณด๋ฉด, ํ˜ˆ์ค‘ ์นผ์Š˜ ๋†๋„๋Š” ๊ทธ๋ฃน ๊ฐ„์˜ ์œ ์˜์ ์ธ ์ฐจ์ด๊ฐ€ ๋‚˜ํƒ€๋‚˜์ง€ ์•Š์•˜๊ณ , ์ธ์˜ ๊ฒฝ์šฐ, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 1.0 g/kg๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ์œ ์˜์  ์ฐจ์ด๋ฅผ ๋ณด์˜€๋‹ค(p < 0.05)). 4, there was no significant difference in blood calcium levels among the groups, and in the case of phosphorus, there was a significant difference in the group administered with 1.0 g / kg of germinated fermented soybeans (p <0.05)).

Alkaline phosphatase(ALP)๋Š” ๊ณจ๋Œ€์‚ฌ์— ๊ด€์—ฌํ•˜๋Š” ์กฐ ๊ณจ์„ธํฌ์˜ biomarker์ด๋‹ค. ๊ณจ๋‹ค๊ณต์ฆ๊ณผ ๊ฐ™์ด ๊ณจ๋Œ€์‚ฌ๊ฐ€ ๊ธ‰๊ฒฉํžˆ ์ฆ๊ฐ€ํ•˜๋Š” ์งˆ๋ณ‘์ด ์žˆ์„ ๊ฒฝ์šฐ, ALP์˜ ์ˆ˜์น˜ ๋˜ํ•œ ์ฆ๊ฐ€ํ•˜๊ฒŒ ๋œ๋‹ค. ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์˜ ํ˜ˆ์ค‘ ALP ์ˆ˜์น˜๋Š” 454.0 ยฑ 69.9 U/L์œผ๋กœ ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์˜ ํ˜ˆ์ค‘ ALP ์ˆ˜ ์น˜ 340.2 ยฑ 33.31 U/L ์— ๋น„ํ•ด ์ฆ๊ฐ€ํ•˜์˜€๋‹ค. ๋ฐ˜๋ฉด, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 0.5 g/kg๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ 260.0 ยฑ 646.9 U /L๋กœ, ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ์œ ์˜์ ์œผ๋กœ ๋‚ฎ์€ ALP ํ™œ์„ฑ์„ ๋ณด์˜€๋‹ค(p < 0.05). ๋˜ํ•œ, ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ๋ฐœ์•„๋ฐœํšจ์ฝฉ 0.5 g/kg๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ Bone-ALP์˜ ์ˆ˜์น˜๊ฐ€ 140.8 ยฑ 58.5 U/L.๋กœ ๋‚˜ํƒ€๋‚˜, ์œ ์˜์ ์œผ๋กœ (p < 0.05) ๋‚ฎ ์•„์ง€๋Š” ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค.Alkaline phosphatase (ALP) is a biomarker of osteoblasts involved in bone metabolism. In other diseases, such as osteoporosis, in which bone metabolism increases rapidly, the level of ALP also increases. The blood ALP level of the ovarian control group was 454.0 ยฑ 69.9 U / L, which was higher than that of the non-ovarian control group (340.2 ยฑ 33.31 U / L). On the other hand, in the group administered 0.5 g / kg of germinated fermented soybeans, 260.0 ยฑ 646.9 U / L, significantly lower ALP activity than the control group (p <0.05). In addition, the Bone-ALP level was 140.8 ยฑ 58.5 U / L. In the group administered with 0.5 g / kg of germinated fermented soybean compared to the control group of ovarian resection, indicating that it was significantly lower (p <0.05). .

๋„ 5๋ฅผ ์‚ดํŽด๋ณด๋ฉด, 6์ฃผ๊ฐ„์˜ ๊ฒฝ๊ตฌํˆฌ์—ฌ๋ฅผ ์‹ค์‹œํ•œ ๋’ค, ๋‚œ์†Œ์ ˆ์ œ ๋Œ€ ์กฐ๊ตฐ์—์„œ ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์™€ ๋น„๊ตํ–ˆ์„ ๋•Œ ์œ ์˜์ ์œผ๋กœ (p < 0.05) ๊ณจ์—ผ๋Ÿ‰์ด ๋‚ฎ์•„์ง€๋Š” ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ ๋‹ค. ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 1.0 g/kg, 2.0 g/kg๋กœ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ๊ฐ๊ฐ์˜ ๊ณจ์—ผ๋Ÿ‰์€ 61.3 ยฑ 0.36%, 60.8 ยฑ 0.31%์œผ๋กœ ๋‚˜ ํƒ€๋‚ฌ๋‹ค. ์ด๋Š” ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ์œ ์˜์ ์œผ๋กœ (p < 0.05) ๊ณจ์—ผ๋Ÿ‰์ด ์ฆ๊ฐ€ํ•œ ์ˆ˜์น˜์ด๋‹ค. ์ด๋ฅผ ํ†ตํ•ด ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ํˆฌ์—ฌ๊ฐ€ ๊ณจ์˜ ๋ฏธ๋„ค๋ž„ ํ•จ๋Ÿ‰์„ ์ฆ๊ฐ€์‹œํ‚ค๋Š” ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค.Referring to FIG. 5, after oral administration for 6 weeks, it was confirmed that the bone inflammation was significantly lower (p <0.05) in the ovarian resection group compared with the non-ovarian resection control group. In the groups administered with 1.0 g / kg and 2.0 g / kg of germinated fermented soybeans, the bone salts were 61.3 ยฑ 0.36% and 60.8 ยฑ 0.31%, respectively. This is a significant (p <0.05) increase in bone salt compared to ovarian resection control. This confirmed that the germination of fermented soybeans increased the mineral content of bone.

2.5. ๊ณจ์กฐ์ง ํ˜•ํƒœ ์ธก์ •: Micro-CT ์ดฌ์˜2.5. Bone Morphology Measurement: Micro-CT

๋‚œ์†Œ์ ˆ์ œ๋ฅผ ํ•œ ๋ชจ๋“  ๊ตฐ์—์„œ ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๊ตฐ์— ๋น„ํ•ด percent bone volume(BV/TV (%)), trabecular number(Tb.N (mm-1)), trabecular thickness(Tb.Th (ฮผm))์˜ ์ˆ˜์น˜๊ฐ€ ๊ฐ์†Œํ•œ ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ๋‹ค. ์—์ŠคํŠธ๋กœ๊ฒ ํˆฌ์—ฌ๊ตฐ๊ณผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 2.0 g/kg ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€ ์กฐ๊ตฐ์— ๋น„ํ•ด ์œ ์˜์ ์œผ๋กœ ๊ทธ ์ˆ˜์น˜๋“ค์ด ์ฆ๊ฐ€ํ•˜์˜€๋‹ค. BV/TV ์˜ ๊ฒฝ์šฐ, ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์—์„œ 13.7 ยฑ 1.3%๋กœ ๊ฐ์†Œํ•˜์˜€์œผ๋ฉฐ, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 2.0 g/kg๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์˜ BT/TV ๊ฐ€ 23.3 ยฑ 1.3% ์œผ๋กœ ๋‚˜ํƒ€๋‚˜, ์œ ์˜์ ์ธ ์ฐจ์ด๋ฅผ ๋ณด์˜€๋‹ค (p < 0.01) (๋„ 6). Tb.N ์˜ ๊ฒฝ์šฐ, ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์—์„œ 1.17 ยฑ 0.09 mm-1๋กœ ๋‚ฎ์€ ์ˆ˜์น˜๋ฅผ ๋ณด์˜€์œผ๋‚˜, ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 2.0 g/kg๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ 1.69 ยฑ 0.15 mm-1๋กœ ์—์ŠคํŠธ๋กœ๊ฒ ํˆฌ์—ฌ๊ตฐ๊ณผ ์œ ์‚ฌํ•œ ์ˆ˜์น˜๋กœ ์ฆ๊ฐ€ํ•œ ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค(p < 0.05). Tb.Th ์˜ ๊ฒฝ์šฐ, ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์—์„œ 0.12 ยฑ 0.00 mm-1๋กœ ๋‚ฎ์€ ์ˆ˜์น˜๋ฅผ ๋ณด์˜€์œผ๋‚˜, ์—์ŠคํŠธ๋กœ๊ฒ ๋ฐ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ๋ชจ๋“  ๋†๋„์—์„œ Tb.Th ๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ•˜์˜€๋‹ค(๋„ 6). ์ด๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ์ฅ์—์„œ ๊ณจ์˜ ๋ฐ€๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ–ˆ๊ณ  , ํ•ด๋ฉด๊ณจ์˜ ๋‘๊ป˜์™€ ์ˆ˜๊ฐ€ ์ฆ๊ฐ€ํ•˜์˜€๋‹ค๋Š” ๊ฒƒ์„ ์˜๋ฏธํ•œ๋‹ค. The values of percent bone volume (BV / TV (%)), trabecular number (Tb.N (mm -1 )), and trabecular thickness (Tb.Th (ฮผm)) in all ovarian sections compared to non-ovarian sections It can be seen that decreases. The estrogen-treated group and the 2.0 g / kg germinated fermented soybean group increased significantly compared to the ovarian ablation control group. BV / TV was decreased to 13.7 ยฑ 1.3% in the control group of ovarian ablation, and BT / TV of the group treated with 2.0 g / kg of germinated fermented soybean was 23.3 ยฑ 1.3%, showing a significant difference (p <0.01 ) (FIG. 6). In the case of Tb.N, the ovarian control group showed a low value of 1.17 ยฑ 0.09 mm -1 , but increased to 1.69 ยฑ 0.15 mm -1 similar to that of the estrogen group in the 2.0 g / kg germinated fermented soybean group. It could be confirmed (p <0.05). In the case of Tb.Th, the ovarian control group showed a low value of 0.12 ยฑ 0.00 mm โˆ’1 , but Tb.Th was significantly increased at all concentrations of estrogen and germinated fermented soybeans (FIG. 6). This means that the bone density of the germinated fermented soybeans increased significantly, and the thickness and number of spongy bones increased.

๋„ 6์„ ๋ณด๋ฉด, Trabecular separation (Tb .Sp (ฮผm)), trabecular pattern factor (Tb.pf (mm-1))์™€ structure model index (SMI)์˜ ๊ฒฝ์šฐ ๋‚œ์†Œ์ ˆ์ œ๋ฅผ ํ•œ ๋ชจ๋“  ๊ตฐ์—์„œ ๋น„ ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ๊ทธ ์ˆ˜์น˜๋“ค์ด ์ฆ๊ฐ€ํ–ˆ๋‹ค. ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 2.0 g/kg ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ Tb.Sp ์ˆ˜์น˜๋Š” 0.70 ยฑ 0.10 ์œผ๋กœ ๋‚˜ํƒ€๋‚˜ ์œ ์˜์ ์œผ๋กœ ๊ฐ์†Œํ•˜์˜€๋‹ค(p < 0.05). ์ด ๊ฒฐ๊ณผ๋ฅผ ํ†ตํ•ด ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ๊ณจ๋‹ค๊ณต์ฆ๊ณผ ๋ฐ€์ ‘ํ•œ ๊ด€๋ จ์ด ์žˆ๋Š” ํ•ด๋ฉด๊ณจ์˜ ๋ถ„์‚ฐ๋„๊ฐ€ ๊ฐ์†Œํ–ˆ์Œ์„ ์•Œ ์ˆ˜ ์žˆ๋‹ค. Tb.pf ์˜ ๊ฒฝ์šฐ, ์—์ŠคํŠธ๋กœ๊ฒ ํˆฌ์—ฌ๊ตฐ๊ณผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 2.0 g/kg ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ๋‚œ์†Œ์ ˆ ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ์œ ์˜์ ์œผ๋กœ (p < 0.01, p < 0.05) ๊ฐ์†Œํ–ˆ๋‹ค. ๊ทธ ์ˆ˜์น˜๋Š” ๊ฐ๊ฐ 0.76 ยฑ 0.48 mm -1, 0.63 ยฑ 1.25 mm-1 ์œผ๋กœ ๋‚˜ํƒ€๋‚ฌ๋‹ค. ๋งˆ์ง€๋ง‰์œผ๋กœ, structure model inde x (SMI)์˜ ๊ฒฝ์šฐ, ์—์ŠคํŠธ๋กœ๊ฒ ํˆฌ์—ฌ๊ตฐ๊ณผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 2.0 g/kg ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ๊ฐ๊ฐ 1.54 ยฑ 0.07 , 1.53 ยฑ 0.10 ์œผ๋กœ, ๋‚œ์†Œ์ ˆ์ œ ๋Œ€์กฐ๊ตฐ์— ๋น„ํ•ด ์œ ์˜์ ์œผ๋กœ(p < 0.01) ๊ฐ์†Œํ–ˆ๋‹ค. ๋”ฐ๋ผ์„œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ 2.0 g/kg ํˆฌ์—ฌ๊ฐ€ ์—์Šค ํŠธ๋กœ๊ฒ ํˆฌ์—ฌ์™€ ๋น„์Šทํ•œ ์ˆ˜์ค€์œผ๋กœ ๊ณจ๋‹ค๊ณต์ฆ์„ ์˜ˆ๋ฐฉํšจ๊ณผ๊ฐ€ ์žˆ์Œ์œผ๋กœ ์˜ˆ์ƒํ•œ๋‹ค. 6, in the case of trabecular separation (Tb.Sp (ฮผm)), trabecular pattern factor (Tb.pf (mm โˆ’1 )) and structure model index (SMI), all groups with ovarian resection were compared to the non-ovarian resection control group. The figures increased. In the 2.0 g / kg group of germinated fermented soybean, the Tb.Sp level was 0.70 ยฑ 0.10, which was significantly decreased (p <0.05). These results show that the degree of dispersal of spongy bone, which is closely related to osteoporosis, was decreased in the group to which the germinated fermented soybeans were administered compared to the ovarian control group. In the case of Tb.pf, estrogen-treated and 2.0 g / kg of germinated fermented soybeans were significantly decreased (p <0.01, p <0.05) compared to the ovarian control group. The figures were 0.76 ยฑ 0.48 mm -1 and 0.63 ยฑ 1.25 mm -1 , respectively. Finally, the structure model inde x (SMI) was 1.54 ยฑ 0.07, 1.53 ยฑ 0.10 in the estrogen-administered group and 2.0 g / kg of germinated fermented soybean, respectively (p <0.01). Decreased. Therefore, the germination fermented soybean 2.0 g / kg administration is expected to be effective in preventing osteoporosis to a level similar to the administration of estrogen.

3. ๊ฐ„ ๋…์„ฑ ๋ชจ๋ธ์˜ ํ•ญ์‚ฐํ™” ํšจ๊ณผ3. Antioxidant Effect of Liver Toxicity Model

3.1. In vitro ํ•ญ์‚ฐํ™” ํ™œ์„ฑ(DPPH, ABTS, Polyphenol)์ธก์ • ๊ฒฐ๊ณผ 3.1. In vitro antioxidant activity (DPPH, ABTS, Polyphenol) measurement results

ํ•˜๊ธฐ ํ‘œ 2๋ฅผ ์‚ดํŽด๋ณด๋ฉด ๊ฐ ์‹œ๋ฃŒ์˜ ํ•ญ์‚ฐํ™” ํ™œ์„ฑ์„ ์ธก์ •ํ•œ ๊ฒฐ๊ณผ ํ•ญ์ƒํ™” ํ™œ์„ฑ์€ ํœด๋ฐ”์ด์˜ค์—์„œ ์ œ๊ณตํ•œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ๋ณด๋‹ค 70% EtOH์„ ์ด์šฉ ํ•˜์—ฌ ์ถ”์ถœํ•œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ํ•ญ์‚ฐํ™” ํ™œ์„ฑ ๋ฐ Polyphenolํ•จ๋Ÿ‰์ด ๋” ๋†’๊ฒŒ ์ธก์ •๋˜์—ˆ๋‹ค.Referring to Table 2 below, the antioxidant activity of each sample was measured, and the homeostasis activity of the germinated fermented soybeans extracted with 70% EtOH was higher than the germinated fermented soybeans provided by Hubio.

In vitro ํ•ญ์‚ฐํ™”ํ™œ์„ฑ(DPPH, ABTS, Polyphenol) ์ธก์ •๊ฒฐ๊ณผIn vitro antioxidant activity (DPPH, ABTS, Polyphenol) measurement results IC50ValuedIC 50 Valued ABTS(mg/ml)ABTS (mg / ml) DPPH(mg/ml)DPPH (mg / ml) Polyphenol(ฮผg/ml)Polyphenol (ฮผg / ml) GFSGFS 4.2936ยฑ0.034.2936 ยฑ 0.03 2.4272ยฑ0.042.4272 ยฑ 0.04 324.09ยฑ9.43324.09 ยฑ 9.43 70%์—ํƒ„์˜ฌ GFS70% Ethanol GFS 2.522ยฑ0.012.522 ยฑ 0.01 0.7366ยฑ0.190.7366 ยฑ 0.19 684.61ยฑ8.94684.61 ยฑ 8.94 ์•„์Šค์ฝœ๋นˆ์‚ฐ(IC50 ฮผg/ml)Ascorbic Acid (IC50 ฮผg / ml) 68.2545ยฑ0.9568.2545 ยฑ 0.95 30.6857ยฑ1.8730.6857 ยฑ 1.87 --

3.2 ์‹คํ—˜ ์žฌ๋ฃŒ (Germinated Fermented and Soybeans : GFS)์˜ ๋ถ„๋ฆฌ ๋ฐ ์ •์ œ3.2 Isolation and Purification of Experimental Materials (Germinated Fermented and Soybeans: GFS)

๋„ 8์„ ์‚ดํŽด๋ณด๋ฉด ํ•ญ์‚ฐํ™” ํ™œ์„ฑ์ด ๋†’๊ฒŒ ์ธก์ •๋œ 70% EtOH์ถ”์ถœ๋ฌผ์„ ์ด์šฉํ•˜์—ฌ Sep-pak์„ ์ด์šฉํ•˜์—ฌ ๋ฌผ์งˆ ๋ถ„๋ฆฌ ํ›„ ํ•ญ์‚ฐํ™” ํ™œ์„ฑ์„ ์ธก์ •ํ•œ ๊ฒฐ๊ณผ Metanol 30%์™€ 70%์—์„œ ๋†’์€ Polyphenol ํ•จ๋Ÿ‰์„ ๋ณด์—ฌ์ฃผ์—ˆ๋‹ค. ๋˜ํ•œ, ํ•˜๊ธฐ ํ‘œ 3์„ ์‚ดํŽด๋ณด๋ฉด ํ•ญ์‚ฐํ™” ํ™œ์„ฑ ์ธก์ • ๋ฐฉ๋ฒ•์ธ ABTS ๊ฒฐ๊ณผ Methanol 70%์— ๊ฐ€์žฅ ๋†’์€ ํ•ญ์‚ฐํ™” ํ™œ์„ฑ์„ ๋‚˜ํƒ€๋‚ด์—ˆ๋‹ค. ํ•˜๊ธฐ ํ‘œ 3์€ In vitro ํ•ญ์‚ฐํ™” ํ™œ์„ฑ(ABTS IC50value, Polyphenol)์ธก์ • ๊ฒฐ๊ณผ๋ฅผ ๋‚˜ํƒ€๋‚ธ ํ‘œ์ด๋‹ค.Referring to FIG. 8, 70% EtOH extract with high antioxidant activity was used to measure the antioxidant activity after separation of the material using Sep-pak. As a result, high polyphenol contents were shown at 30% and 70% of metanol. In addition, looking at the following Table 3 showed the highest antioxidant activity in 70% Methanol results of ABTS results of the antioxidant activity measurement method. Table 3 is a table showing the results of in vitro antioxidant activity (ABTS IC50value, Polyphenol) measurement.

70%์—ํƒ„์˜ฌ GFS ๋ถ„ํš70% Ethanol GFS Fraction 1One 22 33 44 55 66 ABTS(mg/ml)ABTS (mg / ml) 2.96ยฑ0.072.96 ยฑ 0.07 1.16ยฑ0.011.16 ยฑ 0.01 1.09ยฑ0.031.09 ยฑ 0.03 0.68ยฑ0.010.68 ยฑ 0.01 5.62ยฑ0.075.62 ยฑ 0.07 1.09ยฑ0.091.09 ยฑ 0.09 Polyphenol(ฮผg/ml)Polyphenol (ฮผg / ml) 383.1396ยฑ7.61383.1396 ยฑ 7.61 256.7509ยฑ1.63256.7509 ยฑ 1.63 298.9451ยฑ2.65298.9451 ยฑ 2.65 261.6921ยฑ1.73261.6921 ยฑ 1.73 92.8187ยฑ1.5592.8187 ยฑ 1.55 988.1484ยฑ9.67988.1484 ยฑ 9.67

(Ascorbic acid(IC50 ug/ml): 2.13ยฑ0.06)(Ascorbic acid (IC 50 ug / ml): 2.13 ยฑ 0.06)

3.3. HepG2 cell์„ ์ด์šฉํ•œ t-BHP๋กœ ์ธํ•œ ์†์ƒ์œผ๋กœ๋ถ€ํ„ฐ์˜ ์‹œ๋ฃŒ์˜ ๋ณดํ˜ธ ํ™œ์„ฑ ์ธก์ •3.3. Determination of Protective Activity of Samples from Damage Caused by t-BHP Using HepG2 Cells

3.3.1. ์„ธํฌ์ƒ์กด์œจ3.3.1. Cell survival rate

MTT assay๋Š” ๋…ธ๋ž€์ƒ‰์˜ ๊ฐ€์šฉ์„ฑ tetraz olium ์—ผ์ด ์‚ด์•„์žˆ๋Š” ์„ธํฌ์˜ ๋ฏธํ† ์ฝ˜๋“œ๋ฆฌ์•„ succina dehydrogenase์— ์˜ํ•ด ํŒŒ๋ž€์ƒ‰์˜ ๋ถˆ์šฉ์„ฑ formazan ์ƒ์„ฑ๋ฌผ๋กœ ํ™˜์›๋˜๋Š” ์›๋ฆฌ๋Š” ์ด์šฉํ•œ ์‹คํ—˜ ๋ฐฉ๋ฒ•์œผ๋กœ formazan์˜ ํก๊ด‘๋„๋Š” 540 nm์˜ ํŒŒ์žฅ์—์„œ ์ตœ๋Œ€๊ฐ€ ๋œ๋‹ค. The MTT assay is based on the principle that the yellow soluble tetraz olium salt is reduced to the blue insoluble formazan product by mitochondrial succina dehydrogenase in living cells. The absorbance of formazan is maximized at a wavelength of 540 nm.

์ด ํŒŒ์žฅ์—์„œ ์ธก์ •๋œ ํก๊ด‘๋„๋Š” ์‚ด์•„์žˆ๊ณ  ๋Œ€์‚ฌ์ ์œผ๋กœ ์™•์„ฑํ•œ ์„ธํฌ์˜ ๋†๋„๋ฅผ ๋ฐ˜์˜ํ•˜๋Š” ๊ฒƒ์œผ๋กœ ๋ฏธํ† ์ฝ˜๋“œ๋ฆฌ์•„ ํ™œ์„ฑ์˜ ์ง€ํ‘œ๋กœ ์‚ฌ์šฉ๋˜๊ณ  ์žˆ๋‹ค. MTT๋Š” ์„ธํฌ ๋…์„ฑ๋„ ์กฐ์‚ฌ๋กœ ์‹œํ—˜๋ฌผ์งˆ์— ์–ด๋Š ์ •๋„ ๋…์„ฑ์„ ๋‚˜ํƒ€๋‚ด๋Š”์ง€ ๋ฏธํ† ์ฝ˜๋“œ๋ฆฌ์•„์˜ ํ™œ์„ฑ์„ ์ธก์ •ํ•˜๋Š” ์‹คํ—˜์œผ๋กœ ์‹œํ—˜๋ฌผ์งˆ์ด ๋…์„ฑ์ด ์žˆ์–ด ๋ฏธํ† ์ฝ˜๋“œ๋ฆฌ์•„๊ฐ€ ํŒŒ๊ดด๋˜๋ฉด ํก๊ด‘๋„ ๊ฐ’์ด ๋‚ฎ๊ฒŒ ๋‚˜ํƒ€๋‚œ๋‹ค.The absorbance measured at this wavelength reflects the concentration of living and metabolically active cells and is used as an indicator of mitochondrial activity. MTT is a test for measuring the mitochondrial activity to determine how toxic to a test substance by cytotoxicity test. If the test substance is toxic and the mitochondria are destroyed, the absorbance value is low.

๋„ 9A๋ฅผ ๋ณด๋ฉด, HepG2 cell์— ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์‹œ๋ฃŒ๋งŒ ์ฒ˜๋ฆฌํ•œ ๊ฒฝ์šฐ์—๋Š” ์‹œ๋ฃŒ๋ฅผ ์ฒ˜๋ฆฌํ•˜์ง€ ์•Š์€ ์„ธํฌ(100%) ๋น„๊ตํ•˜์˜€์„ ๋•Œ ๋ฐœ์•„๋ฐœํšจ ์ฝฉ์„ ์ฒ˜๋ฆฌํ•œ ๊ตฐ์—์„œ 0.0312(98.52 ยฑ 3.64), 0.0625(97.08 ยฑ 2.93), 0.125(94.44 ยฑ 1.72), 0.25(96.46 ยฑ 2.86), 0.5(96.19 ยฑ 3.03)mg/mL ๋†๋„์—์„œ ์œ ์˜์ ์œผ๋กœ ์•ฝ๊ฐ„์˜ ๋…์„ฑ์„ ๋‚˜ํƒ€๋‚ด์—ˆ๋‹ค.ย 9A shows that when HepG2 cells were treated with only germinated fermented soybean samples, 0.0312 (98.52 ยฑ 3.64), 0.0625 (97.08 ยฑ 2.93) in the group treated with germinated fermented soybeans when compared with the untreated cells (100%). ), 0.125 (94.44 ยฑ 1.72), 0.25 (96.46 ยฑ 2.86), and 0.5 (96.19 ยฑ 3.03) mg / mL concentrations showed significant toxicity.

๋˜ํ•œ, ๋„ 9B๋ฅผ ๋ณด๋ฉด 70% EtoH ์ถ”์ถœ๋ฌผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ์ด์šฉํ•˜์—ฌ ์„ธํฌ์— ์ฒ˜๋ฆฌํ•˜์˜€์„ ๋•Œ์˜ ์„ธํฌ์ƒ์กด์œจ์€ ์‹œ๋ฃŒ๋ฅผ ์ฒ˜๋ฆฌํ•˜์ง€ ์•Š์€ ๊ตฐ๊ณผ ์ฐจ์ด๊ฐ€ ์—†์—ˆ๋‹ค.In addition, as shown in Figure 9B the cell survival rate when the cells were treated with 70% EtoH extract germinated fermented soybean did not differ from the group not treated with the sample.

์•„์šธ๋Ÿฌ, ๋„ 10์˜ A๋ฅผ ๋ณด๋ฉด, HepG2 cell์„ ์ด์šฉํ•˜์—ฌ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ๊ณผ t-BHP๋ฅผ ์ฒ˜๋ฆฌํ•œ ๊ฒฝ์šฐ์—๋Š” ์•„๋ฌด๊ฒƒ๋„ ์ฒ˜๋ฆฌํ•˜์ง€ ์•Š์€ ์„ธํฌ์™€ ๋น„๊ตํ•˜์˜€์„ ๋•Œ, t-BHP๋งŒ ์ฒ˜๋ฆฌํ•œ ์„ธํฌ์˜ ๊ฒฝ์šฐ์—๋Š” ์•ฝ60% ์ •๋„ ๋‚ฎ๊ฒŒ ๋‚˜์™”์œผ๋ฉฐ t-BHP๋งŒ ์ฒ˜๋ฆฌํ•œ ์„ธํฌ์™€ ๋น„๊ตํ•˜์˜€์„ ๋•Œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์‹œ๋ฃŒ๋ฅผ ์ฒ˜๋ฆฌํ•˜์˜€์„ ๋•Œ๋Š” t-BHP๋งŒ ์ฒ˜๋ฆฌํ•œ ์„ธํฌ์™€ ํฐ ์ฐจ์ด๋ฅผ ๋ณผ ์ˆ˜ ์—†์—ˆ๋‹ค. ํ•˜์ง€๋งŒ, ๋„ 10B๋ฅผ ์‚ดํŽด๋ณด๋ฉด, 70%EtoH ์ถ”์ถœ๋ฌผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ t-BHP๋งŒ ์ฒ˜๋ฆฌํ•œ ์„ธํฌ(64.67 ยฑ 0.7 8%)์™€ ๋น„๊ตํ•˜์˜€์„ ๋•Œ๋Š” ๋†๋„ ์œ ์˜์ ์ด์ง€๋Š” ์•Š์ง€๋งŒ ์„ธํฌ์˜ ์ƒ์กด์œจ์ด ์ฆ๊ฐ€ํ•˜๋Š” ๊ฒƒ์„ ํ™•์ธํ•  ์ˆ˜ ์žˆ๋‹ค. 0.0625( 80.11 ยฑ 0.90%), 0.125(82.67 ยฑ 0.84%), 0.25(83.52 ยฑ1.98%), 0.5(82.54 ยฑ 2.58%)mg/mL.In addition, as shown in Figure 10A, when treated with germinated fermented soybeans and t-BHP using HepG2 cells, compared to cells that did not process anything, about 60% of cells treated with t-BHP only When compared with the cells treated with t-BHP alone, the germinated fermented soybean samples were not significantly different from those treated with t-BHP alone. However, referring to FIG. 10B, when the 70% EtoH extract germinated fermented soybean is compared with the cells treated with t-BHP only (64.67 ยฑ 0.7 8%), the concentration of the cells is not significant but it can be seen that the survival rate of the cells increases. 0.0625 (80.11 ยฑ 0.90%), 0.125 (82.67 ยฑ 0.84%), 0.25 (83.52 ยฑ 1.98%), 0.5 (82.54 ยฑ 2.58%) mg / mL.

3.4. HepG2 cell์„ ์ด์šฉํ•œ ROS(reactive oxygen species) ์ธก์ • 3.4. Reactive oxygen species (ROS) measurement using HepG2 cells

DCFH-DA (dichlorofluorescin diacetate)๋Š” ์„ธํฌ๋‚ด์˜ ํ™œ์„ฑ์‚ฐ์†Œ (ROS)์™€ ๋ฐ˜์‘ํ•˜์—ฌ ๊ฐ•ํ•œ ํ˜•๊ด‘๋ฌผ์งˆ์ธ DCF (dichlorofluorescein)๋กœ ์ „ํ™˜๋˜๊ฒŒ ๋œ๋‹ค. ๋”ฐ๋ผ์„œ DCF์— ์˜ํ•œ ํ˜•๊ด‘๋„๋ฅผ ์ธก์ •ํ•จ์œผ๋กœ์จ ROS์˜ ์ƒ๋Œ€์ ์ธ ๋Ÿ‰์„ ์ถ”์ธกํ•  ์ˆ˜ ์žˆ๋‹ค. Dichlorofluorescin diacetate (DCFH-DA) reacts with free radicals (ROS) in the cell and converts it into a strong fluorescent substance, dichlorofluorescein (DCF). Therefore, the relative amount of ROS can be estimated by measuring the fluorescence by DCF.

70%EtoH ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์— ๋Œ€ํ•œ ROS ํšจ๊ณผ๋ฅผ ํ™•์ธํ•˜๊ธฐ ์œ„ํ•˜์—ฌ 70%EtoH ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์„ธํฌ๋…์„ฑ์ด ์—†๋Š” ๋†๋„๋กœ 12์‹œ๊ฐ„ ๋™์•ˆ ๋ฐฐ์–‘ํ•˜๊ณ  t-BHP 1mM์„ 1์‹œ๊ฐ„ ๋™์•ˆ ์ฒ˜๋ฆฌํ•˜์—ฌ ์‚ฐํ™”์  ์ŠคํŠธ๋ ˆ์Šค๋ฅผ ์œ ๋ฐœํ•˜์˜€๋‹ค. ์—ฌ๊ธฐ์— 25 ฮผM์˜ DCFH-DA๋ฅผ 60๋ถ„ ๋™์•ˆ ๋„ฃ์€ ํ›„์— DCF์˜ ํ˜•๊ด‘๋„๋ฅผ ์ธก์ •ํ•˜์˜€๋‹ค. In order to confirm the ROS effect on 70% EtoH germinated fermented soybean extract, 70% EtoH germinated fermented soybean extract was incubated for 12 hours in a non-cytotoxic concentration and t- BHP 1 mM was treated for 1 hour to induce oxidative stress. . After 25 ฮผM of DCFH-DA was added for 60 minutes, the fluorescence of DCF was measured.

๊ทธ ๊ฒฐ๊ณผ๋Š” ๋„ 11๊ณผ ๊ฐ™๋‹ค. ๋„ 11์„ ์‚ดํŽด๋ณด๋ฉด, Normal(100 ยฑ 1.00%)์— ๋น„ํ•ด t-BHP ์ฒ˜๋ฆฌ์— ์˜ํ•ด (267.62 ยฑ 2.68%)๋กœ ํ˜•๊ด‘๋„๊ฐ€ ์ฆ๊ฐ€ ํ•˜์˜€๊ณ , 70%EtoH ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ ์ „์ฒ˜๋ฆฌ์— ์˜ํ•ด์„œ 0.5(195.24 ยฑ 1.95%), 0.25mg/mL(195.24 ยฑ 1.95), 0.125mg/mL(240 ยฑ 2.41), 0.0625mg/mL (252.80 ยฑ 2.53)๋กœ ๊ฐ€์žฅ ๋†’ ์€ ๋†๋„๋ถ€ํ„ฐ ๋†๋„ ์œ ์˜์ ์œผ๋กœ ์–ต์ œ๋˜์–ด 70% EtoH ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์— ์˜ํ•œ ํ™œ์„ฑ์‚ฐ์†Œ ๊ฐ์†Œํšจ๊ณผ๋ฅผ ํ™•์ธํ•  ์ˆ˜ ์žˆ์—ˆ๋‹ค.The result is shown in FIG. Referring to Figure 11, compared with Normal (100 ยฑ 1.00%) fluorescence increased by (267.62 ยฑ 2.68%) by t-BHP treatment, 0.5 (195.24 ยฑ 1.95%) by 70% EtoH germinated fermented soybean extract pretreatment , 0.25mg / mL (195.24 ยฑ 1.95), 0.125mg / mL (240 ยฑ 2.41), 0.0625mg / mL (252.80 ยฑ 2.53), significantly inhibited the concentration from the highest concentration to 70% EtoH germinated fermented soybean extract It was confirmed that the active oxygen reduction effect by.

3.5. ๋™๋ฌผ๋ชจ๋ธ์„ ์ด์šฉํ•œ ํ˜ˆ์•กํ•™์  ๊ฒ€์‚ฌ3.5. Hematological tests using animal models

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•˜๊ณ  t-BHPํˆฌ์—ฌํ•œ ๊ตฐ์˜ ํ˜ˆ์•ก ์ƒํ™”ํ•™์  ๊ฒ€์‚ฌ๊ฒฐ๊ณผ๋Š” ์•„๋ž˜์˜ ํ‘œ 4์— ๋‚˜ํƒ€๋‚ด์—ˆ๋‹ค. Total Cholestrol, LDH, Total Bilirubin, ALP, ALT, ์ธก์ •ํ•œ ๊ฒฐ๊ณผ ๋Œ€์กฐ๊ตฐ์— ๋น„๊ตํ•˜์˜€์„ ๋•Œ ์œ ์˜์ ์ธ ์ฐจ์ด(ํ‘œ 4)๊ฐ€ ๋‚˜ํƒ€๋‚ฌ๋‹ค. LDH๋Š” ํ•ด๋‹น๊ณ„ ํšจ์†Œ๋กœ์„œ ์ฒด๋‚ด์˜ ๊ฐ ์กฐ์ง์— ๋„๋ฆฌ ๋ถ„ํฌ๋˜์–ด ์žˆ์œผ๋ฉฐ, ํŠนํžˆ ์‹ฌ์žฅ, ๊ฐ„์žฅ, ์‹ ์žฅ ๋ฐ ๊ทผ์œก์— ์กด์žฌํ•˜๋Š” ํ˜ˆ ์ฒญ์ค‘ LDH๋Š” ์‹ฌ์งˆํ™˜, ๊ฐ„์งˆํ™˜ ์•…์„ฑ์ข…์–‘ ๋ฐ ๋ฐฑํ˜ˆ๋ณ‘ ๋“ฑ์—์„œ ์ฆ๊ฐ€ํ•˜๋‚˜ ํŠน์ด์„ฑ์ด ๋‚ฎ๋‹ค. ๊ธ‰์„ฑ๊ฐ„์—ผ์—์„œ๋Š” AST, ALT์™€ ๋™์ผํ•˜๊ฒŒ ๊ฐ„์„ธํฌ๋กœ๋ถ€ํ„ฐ ๋น ์ ธ๋‚˜์™€์„œ ๊ธ‰์„ฑ๊ฐ„์—ผ์ดˆ๊ธฐ์— ์ฆ๊ฐ€๋˜๋ฉฐ ๋งŒ์„ฑ๊ฐ„์—ผ ๋ฐ ๊ฐ„๊ฒฝ๋ณ€์ฆ์—์„œ๋Š” ์ •์ƒ์ด๊ฑฐ๋‚˜ ๊ฒฝ๋„๋กœ ์ฆ๊ฐ€ํ•œ๋‹ค. ๊ฐ„์žฅ์• ๋ฅผ ๋‚˜ํƒ€๋‚ด๋Š” ์ง€ํ‘œ ์ค‘์˜ ํ•˜๋‚˜์ธ AST์™€ ALT์—์„œ ์œ ์˜์ ์ธ ์ฐจ์ด๊ฐ€ ๋‚˜ํƒ€๋‚ฌ๋‹ค.Blood biochemical test results of the group administered with germinated fermented soybean and administered with t-BHP are shown in Table 4 below. Total Cholestrol, LDH, Total Bilirubin, ALP, ALT, and measurement results showed a significant difference (Table 4) when compared to the control group. LDH is a glycolytic enzyme and widely distributed in each tissue in the body. Especially, LDH in serum, liver, kidney, and muscle increases in heart disease, liver disease, malignancy, and leukemia, but has low specificity. In acute hepatitis, it escapes from hepatocytes like AST and ALT and increases in early acute hepatitis. In chronic hepatitis and cirrhosis, it increases to normal or mild. Significant differences were found in AST and ALT, one of the indicators of liver failure.

ALT์™€ AST๋Š” ๊ฐ„์„ธํฌ ๋ˆ„์ถœํšจ์†Œ๋กœ ๊ฐ„์„ธํฌ์˜ ๊ดด์‚ฌ์™€ ๊ฐ„ ์กฐ์ง์˜ ํŒŒ๊ดด๊ฐ€ ์ง„ํ–‰๋จ์— ๋”ฐ๋ผ ํšจ์†Œ๊ฐ€ ํ˜ˆ์ค‘์œผ๋กœ ์œ ๋ฆฌ๋˜์–ด ๋‚˜ํƒ€๋‚˜๋ฏ€๋กœ ๊ฐ„๋…์„ฑ ์—ฐ๊ตฌ์— ํ™œ์šฉ๋˜๊ณ  ์žˆ๋‹ค. ๋„ 12๋Š” ALT์™€ ASTํ™œ์„ฑ์„ ์ธก์ •ํ•œ ๊ฒฐ๊ณผ๋‹ค. AST๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ๋†๋„๊ฐ€ 0.5 g/kg(83.20 ยฑ 2.86 U/L), 1g/kg(81.80 ยฑ 6.72U/L)๋กœ ๋†๋„ ์œ ์˜์  ๋ณ€ํ™”๋Š” ์—†์—ˆ์ง€๋งŒ, ์‹œ๋ฃŒ ์ฒ˜๋ฆฌ๊ตฐ์—์„œ t-BHP๋งŒ ์ฒ˜๋ฆฌํ•œ ๊ตฐ(94.25 ยฑ 7.37 U/L)๊ณผ ๋น„๊ตํ•˜์˜€์„ ๋•Œ ์œ ์˜์ ์œผ๋กœ ๋‚ฎ์•„์กŒ๋‹ค. ALT๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ๋†๋„๊ฐ€ 0.5g/kg(33.71 ยฑ 5.59 U/L), 1g/kg(38.17 ยฑ 4.49 U/L)๋กœ ๋†๋„ ์œ ์˜์  ๋ณ€ํ™”๋Š” ์—†์—ˆ์ง€๋งŒ, ์‹œ๋ฃŒ ์ฒ˜๋ฆฌ๊ตฐ์—์„œ t-BHP๋งŒ ์ฒ˜๋ฆฌํ•œ ๊ตฐ(48.25 ยฑ 16.19 U/L)๊ณผ ๋น„๊ตํ•˜์˜€์„ ๋•Œ ์œ ์˜์ ์œผ๋กœ ๋‚ฎ์•„์กŒ๋‹ค.ย ALT and AST are hepatocellular leak enzymes, which are used for hepatotoxicity studies because enzymes are released into the blood as hepatic necrosis and hepatic tissue destruction progress. 12 shows the results of measuring ALT and AST activity. The concentration of AST was 0.5 g / kg (83.20 ยฑ 2.86 U / L) and 1 g / kg (81.80 ยฑ 6.72 U / L), but there was no significant change in the concentration of germinated fermented soybeans. Significantly lower compared to the group (94.25 ยฑ 7.37 U / L). The ALT showed no significant change in the concentration of germinated fermented soybean at 0.5 g / kg (33.71 ยฑ 5.59 U / L) and 1 g / kg (38.17 ยฑ 4.49 U / L), but only the t-BHP was treated in the sample treatment group. Significantly lower compared to (48.25 ยฑ 16.19 U / L).

TreatmentTreatment TCHO(mg/dl)TCHO (mg / dl) LDH(U/Liter)LDH (U / Liter) TBIL(mg/dl)TBIL (mg / dl) NormalNormal 80.00ยฑ7.81abc 80.00 ยฑ 7.81 abc 123.33ยฑ21.83b 123.33 ยฑ 21.83 b 0.48ยฑ0.05ab 0.48 ยฑ 0.05 ab t-BHP Controlt-BHP Control 85.25ยฑ3.77a 85.25 ยฑ 3.77 a 227.0ยฑ46.66a 227.0 ยฑ 46.66 a 0.60ยฑ0.22a 0.60 ยฑ 0.22 a SilymarinSilymarin 80.8ยฑ6.87ab 80.8 ยฑ 6.87 ab 141.33ยฑ24.34b 141.33 ยฑ 24.34 b 0.54ยฑ0.05ab 0.54 ยฑ 0.05 ab GFS 0.5g/kg+t-BHPGFS 0.5g / kg + t-BHP 66.29ยฑ9.89bc 66.29 ยฑ 9.89 bc 127.0ยฑ27.38b 127.0 ยฑ 27.38 b 0.49ยฑ0.07bc 0.49 ยฑ 0.07 bc GFS 1g/kg+t-BHPGFS 1g / kg + t-BHP 64.63ยฑ7.89c 64.63 ยฑ 7.89 c 102.20ยฑ16.83b 102.20 ยฑ 16.83 b 0.41ยฑ0.04c 0.41 ยฑ 0.04 c

3.6. ๋™๋ฌผ๋ชจ๋ธ์„ ์ด์šฉํ•œ MDA์ธก์ •๊ฒฐ๊ณผ3.6. MDA measurement results using animal models

์ง€์งˆ ๊ณผ์‚ฐํ™”๋Š” ์ƒ์ฒด์—์„œ ์ž์—ฐ์ ์œผ๋กœ ๋ฐœ์ƒ๋  ์ˆ˜ ์žˆ๋Š” superoxide anion radical๋กœ๋ถ€ํ„ฐ ์œ ๋ž˜ํ•˜๋Š” ๊ฐ์ข…์˜ free radical์— ์˜ํ•ด ์„ธํฌ๋ง‰์˜ ๋‹ค๊ฐ€ ๋ถˆํฌํ™”์ง€๋ฐฉ์‚ฐ์ด ๊ณผ์‚ฐํ™”๋˜๋Š” ํ˜„์ƒ์„ ์ง€์นญํ•œ๋‹ค. ๋ถˆํฌํ™”์ง€๋ฐฉ์‚ฐ์€ ๊ณผ์‚ฐํ™” ๊ณผ์ •์„ ํ†ตํ•ด ๋ถ„ํ•ด๋˜์–ด MDA๋ฅผ ์ƒ์„ฑํ•˜๋ฏ€๋กœ ์ด๋ฅผ ์ง€์งˆ๊ณผ์‚ฐํ™”๋ฅผ ์ธก์ •ํ•˜๋Š” ์ง€ํ‘œ๋กœ ์‚ผ๋Š”๋‹ค. ๋„ 13์„ ๋ณด๋ฉด, ๊ฐ„์กฐ์ง์˜ ๊ฒฝ์šฐ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์€ ์‹œ๋ฃŒํˆฌ์—ฌ๋†๋„ 0.5g/kg(34.25 ยฑ 5.80), 1g/kg(32.68 ยฑ 12.57) ์—์„œ ๊ฐ ๊ตฐ๊ฐ„์˜ ์œ ์˜์ ์ธ ์ฐจ์ด๋Š” ์—†์—ˆ์œผ๋‚˜ ๋†๋„๊ฐ€ ๋†’์•„์งˆ์ˆ˜๋ก ๊ฐ์†Œํ•˜์˜€์œผ๋ฉฐ t-BHP๋งŒ ํˆฌ์—ฌํ•œ ๊ตฐ(44.66 ยฑ 1.94)๊ณผ ๋น„๊ตํ•˜์˜€์„ ๋•Œ ๊ฐ์†Œํ•˜๋Š” ๊ฒฝํ–ฅ์„ ๋ณด์˜€๋‹ค.Lipid peroxidation refers to a phenomenon in which polyunsaturated fatty acids in cell membranes are peroxidated by various free radicals derived from superoxide anion radicals that can occur naturally in a living body. Unsaturated fatty acids are degraded through peroxidation to produce MDA, which is an indicator of lipid peroxidation. 13, in the case of liver tissue, the group administered t- BHP after the administration of germinated fermented soybeans was significantly different between the groups at the sample concentration of 0.5 g / kg (34.25 ยฑ 5.80) and 1 g / kg (32.68 ยฑ 12.57). There was no difference, but it decreased as the concentration increased and decreased compared to the t- BHP-only group (44.66 ยฑ 1.94).

3.7. ๋™๋ฌผ๋ชจ๋ธ์„ ์ด์šฉํ•œ GST ์ธก์ •๊ฒฐ๊ณผ3.7. GST measurement results using animal models

GST๋Š” ์‹ ์ฒด์˜ ๊ฑฐ์˜ ๋ชจ๋“  ์„ธํฌ์— ์กด์žฌํ•˜๊ณ  ๊ฐ„์˜ ํ•ด๋…์ž‘์šฉ๋ฟ๋งŒ ์•„๋‹ˆ๋ผ ๋งŽ์€ ์‹ ์ง„๋Œ€์‚ฌ์— ๊ด€์—ฌ ํ•˜๋Š” ํšจ์†Œ์ด๋‹ค. ํŠนํžˆ GST๋Š” ์ „์ž์นœํ™”์„ฑ ๊ธฐ์งˆ์„ GSH์— ๊ฒฐํ•ฉ์‹œํ‚ค๋Š” ๋ฐ˜์‘์„ ์ด‰๋งคํ•˜๋Š” ์—ญํ• ์„ ํ•˜๊ฒŒ๋œ๋‹ค. ๋„ 14๋ฅผ ๋ณด๋ฉด, GST์ธก์ • ๊ฒฐ๊ณผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ 0.5g/kg, 1g/kg, t-BHP๋งŒ ํˆฌ์—ฌํ•œ ๊ตฐ (25.68 ยฑ 4.28)๊ณผ ๋น„๊ตํ•˜์˜€์„ ๋•Œ ์‹œ๋ฃŒํˆฌ์—ฌ๋†๋„ 0.5g/kg (33.65 ยฑ 12.97), 1g/kg (48.43 ยฑ 8.90) ์—์„œ ์‹œ ๋ฃŒ๋†๋„๊ฐ€ ๋†’์•„์งˆ์ˆ˜๋ก ์ฆ๊ฐ€ํ•˜์˜€์œผ๋‚˜ t-BHPํˆฌ์—ฌํ•œ ๊ตฐ๊ณผ์˜ ์œ ์˜์ ์ธ ์ฐจ์ด๋Š” 1g/kg์—์„œ๋งŒ ๋‚˜ํƒ€๋‚ฌ๋‹ค.GST is an enzyme that exists in almost every cell of the body and is involved in many metabolism as well as detoxification of the liver. In particular, GST catalyzes the reaction of binding an electron-affinity substrate to GSH. Referring to FIG. 14, GST measurements germination fermentation after administration of the beans treated group, t -BHP 0.5g / kg, 1g / kg, only one group administered t -BHP (25.68 ยฑ 4.28) Sample administration as compared to the concentration At 0.5 g / kg (33.65 ยฑ 12.97) and 1 g / kg (48.43 ยฑ 8.90), the sample concentration increased with increasing sample concentration, but significant difference with the t-BHP group was only observed at 1 g / kg.

3.8. ๋™ ๋ฌผ๋ชจ๋ธ์„ ์ด์šฉํ•œ GSH ์ธก์ •๊ฒฐ๊ณผ3.8. GSH measurement result using animal model

GSH๋Š” ์ฒด๋‚ด์—์„œ ํ•ด๋…๊ธฐ๋Šฅ, ๋ฉด์—ญ๊ธฐ๋Šฅ, ํ•ญ์‚ฐํ™” ๊ธฐ๋Šฅ๋“ฑ์˜ ํ•ต์‹ฌ์ ์ธ ์—ญํ• ์„ ํ•˜๋Š” ์ธ์ฒด์—์„œ๋Š” ๋งค์šฐ ์ค‘์š”ํ•œ ํ•ด๋…์ œ์ด๋ฉฐ ๊ฐ•๋ ฅํ•œ ํ•ญ์‚ฐํ™”์ œ์ด๋‹ค. ๋„ 13์„ ๋ณด๋ฉด, GSH์ธก์ • ๊ฒฐ๊ณผ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ 0.5g/kg, 1g/kg, t-BHP๋งŒ ํˆฌ์—ฌํ•œ ๊ตฐ21.07 ยฑ 8.33)๊ณผ ๋น„๊ตํ•˜์˜€์„ ๋•Œ ์‹œ๋ฃŒํˆฌ์—ฌ๋†๋„ 0.5g/kg (25.12 ยฑ 2.44), 1g/kg (24.40 ยฑ 2.52) ์—์„œ ์‹œ๋ฃŒ๋†๋„๊ฐ€ ๋†’์•„์งˆ์ˆ˜๋ก ์œ ์˜์  ์ฆ๊ฐ€๋Š” ์—†์—ˆ๋‹ค. ๋˜ํ•œ t-BHPํˆฌ์—ฌํ•œ ๊ตฐ๊ณผ์˜ ์‹œ๋ฃŒ๋ฅผ ํˆฌ์—ฌํ•œ๊ตฐ ๊ฐ„์˜ ์œ ์˜์  ์ฐจ์ด๋Š” ๋ณด์ด์ง€ ์•Š์•˜์ง€๋งŒ, ์‹œ๋ฃŒ๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ์•ฝ๊ฐ„์˜ ์ฆ๊ฐ€ํ•˜๋Š” ๊ฒฝํ–ฅ์„ ๋ณด์—ฌ์ฃผ์—ˆ๋‹ค.GSH is a very important antidote and powerful antioxidant in the human body that plays a key role in detoxification, immune function, and antioxidant function in the body. Referring to FIG. 13, GSH measurements germination fermentation after administration of the beans treated group, t -BHP 0.5g / kg, 1g / kg, t -BHP only administered group 21.07 ยฑ 8.33) Sample administration concentration of 0.5 as compared to the There was no significant increase as the sample concentration increased at g / kg (25.12 ยฑ 2.44) and 1 g / kg (24.40 ยฑ 2.52). In addition, there was no significant difference between the groups receiving the sample and the t- BHP group, but there was a slight increase in the group receiving the sample.

3.9. ๋™๋ฌผ๋ชจ๋ธ์„ ์ด์šฉํ•œ SOD ์ธก์ •๊ฒฐ๊ณผ3.9. SOD measurement results using animal models

SOD๋Š” superoxide anion(O2)๋ฅผ H2O2๋กœ ํ™˜์›์‹œํ‚ค๋Š” ์ž‘์šฉ์„ ํ•˜๋Š” ํšจ์†Œ๋กœ์„œ, cytosol๊ณผ mitochondria์— ์กด์žฌํ•œ๋‹ค. Cytosol์—๋Š” Cu/Zn-SOD๋งŒ์ด ์กด์žฌํ•˜๋‚˜ mitochondria์˜ matrix์—๋Š” Mn-SOD๊ฐ€, ๋‚ด๋ง‰๊ณผ ์™ธ๋ง‰ ์‚ฌ์ด์—๋Š” Cu/Zn-SOD๊ฐ€ ์†Œ๋Ÿ‰ ์กด์žฌ ํ•œ๋‹ค. SOD์ธก์ • ๊ฒฐ๊ณผ(๋„ 9) ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t-BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ 0.5g/kg, 1g/kg, t-BHP๋งŒ ํˆฌ์—ฌํ•œ ๊ตฐ(23.11 ยฑ 1.71)๊ณผ ๋น„๊ตํ•˜์˜€์„ ๋•Œ ์‹œ๋ฃŒํˆฌ์—ฌ๋†๋„ 0.5g/kg (27.35 ยฑ 3.32), 1g/kg (25 .67 ยฑ 0.02) ์—์„œ ์‹œ๋ฃŒ๋†๋„๊ฐ€ ๋†’์•„์งˆ์ˆ˜๋ก ์œ ์˜์  ์ฆ๊ฐ€๋Š” ์—†์—ˆ๋‹ค. ๋˜ํ•œ t-BHPํˆฌ์—ฌํ•œ ๊ตฐ๊ณผ์˜ ์‹œ๋ฃŒ๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ๊ฐ„์˜ ์œ ์˜์  ์ฐจ์ด๊ฐ€ ์žˆ๋‹ค. ๋‹ค๋งŒ, ์‹œ๋ฃŒ๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ๋†๋„์˜์กด์ ์ธ ํšจ๊ณผ๋ฅผ ๋ณด์—ฌ์ฃผ์ง€๋Š” ์•Š์•˜๋‹ค.SOD is an enzyme that reduces superoxide anion (O 2 ) to H 2 O 2 , and is present in cytosol and mitochondria. Only Cu / Zn-SOD exists in the cytosol, but Mn-SOD exists in the matrix of mitochondria, and a small amount of Cu / Zn-SOD exists between the inner and outer layers. SOD measurement results (Fig. 9) germination fermentation after administration of the beans when compared with the group (23.11 ยฑ 1.71) of administration of only the group treated with t -BHP 0.5g / kg, 1g / kg, t -BHP administration sample concentration 0.5 There was no significant increase as the sample concentration increased at g / kg (27.35 ยฑ 3.32) and 1 g / kg (25 .67 ยฑ 0.02). There is also a significant difference between the group administered with the sample and the group administered with t- BHP. However, there was no concentration-dependent effect in the group administered the sample.

3.10. ๋™๋ฌผ๋ชจ๋ธ์„ ์ด์šฉํ•œ CAT ์ธก์ •๊ฒฐ๊ณผ3.10. CAT measurement results using animal model

Catalase๋Š” SOD์˜ ์ž‘์šฉ์ด๋‚˜ O2์˜ ํ™˜์›์œผ๋กœ ์ƒ์„ฑ๋œ H2O2๋ฅผ ๋ฌด๋…์„ฑ์˜ H2O๋กœ ํ™˜์›์‹œ์ผœ ์‚ฐ์†Œ์˜ ๋…์œผ๋กœ๋ถ€ํ„ฐ ์ƒ์ฒด๋ฅผ ๋ณดํ˜ธํ•˜๋Š” ํšจ์†Œ๋กœ์„œ ๋Œ€๋ถ€๋ถ„ peroxisome์— ์กด์žฌํ•˜๊ณ  mitochondria์—๋„ ์†Œ๋Ÿ‰ ์กด์žฌํ•œ๋‹ค. CAT์ธก์ • ๊ฒฐ๊ณผ(๋„ 10) ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ํˆฌ์—ฌํ•œ ํ›„ t -BHP๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ 0.5g/kg, 1g/kg, t-BHP๋งŒ ํˆฌ์—ฌํ•œ ๊ตฐ(182.24 ยฑ 19.29)๊ณผ ๋น„๊ตํ•˜์˜€์„ ๋•Œ ์‹œ๋ฃŒํˆฌ์—ฌ๋†๋„ 0.5g/kg (204.49 ยฑ 41.51), 1g/kg (293.19 ยฑ 61.02) ์—์„œ ์‹œ๋ฃŒ๋†๋„๊ฐ€ ๋†’์•„์งˆ์ˆ˜๋ก ์œ ์˜์  ์ฆ๊ฐ€ํ•˜์˜€๋‹ค. ๋˜ํ•œ t-BHPํˆฌ์—ฌํ•œ ๊ตฐ๊ณผ์˜ ์‹œ๋ฃŒ๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ๊ฐ„์˜ ์œ ์˜์  ์ฐจ์ด๊ฐ€ ์žˆ์œผ๋ฉฐ, ์‹œ๋ฃŒ๋ฅผ ํˆฌ์—ฌํ•œ ๊ตฐ์—์„œ ๋†๋„์˜์กด์ ์ธ ํšจ๊ณผ๋ฅผ ๋ณด์—ฌ์ฃผ์—ˆ๋‹ค.Catalase is an enzyme that protects the body from oxygen poison by reducing H 2 O 2 produced by the action of SOD or O 2 reduction to non-toxic H 2 O. Mostly, it is present in peroxisome and in small amounts in mitochondria. CAT measurement results (Fig. 10) after germination fermented beans treated with the group treated with t -BHP 0.5g / kg, 1g / kg, t -BHP only one dose group (182.24 ยฑ 19.29) administration sample concentration of 0.5 as compared to the At g / kg (204.49 ยฑ 41.51) and 1 g / kg (293.19 ยฑ 61.02), the sample concentration increased significantly. In addition, there was a significant difference between the group administered with the sample and the t- BHP group, and the group administered with the sample showed a concentration-dependent effect.

<์ ์šฉ์˜ˆ> <Application example>

๋ณธ ๋ฐœ๋ช…์€ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์˜ ์ถ”์ถœ๋ฌผ ์ด์™ธ์— ์•ฝ๋ฆฌํ•™์ , ๊ฑด๊ฐ•๊ธฐ๋Šฅ์‹ํ’ˆ๋ฒ•, ์‹ํ’ˆ์œ„์ƒ๋ฒ•์ƒ์œผ๋กœ ํ—ˆ์šฉ๋˜๋Š” ๋ถ€ํ˜•์ œ(๋ถ•ํ•ด์ œ, ๊ฒฐํ•ฉ์ œ, ํ™œํƒ์ œ), ์•ˆ์ •์ œ, ์œ ํ™”์ œ, ์ฒจ๊ฐ€๋ฌผ ๋“ฑ์œผ๋กœ ์ด๋ฃจ์–ด์ง€๋Š” ๊ตฐ์—์„œ 1์ข… ์ด์ƒ ์„ ํƒ๋œ ๊ฒƒ์„ ํฌํ•จํ•  ์ˆ˜ ์žˆ๋‹ค. ๋˜ํ•œ, ์ƒ๊ธฐ ์กฐ์„ฑ๋ฌผ์€ ๊ฒฝ๊ณ ์ œ, ๊ณผ๋ฆฝ์ œ, ์‚ฐ์ œ, ์ •์ œ, ๋˜๋Š” ์บก์Š์ œ ๋“ฑ์˜ ๊ฒฝ๊ตฌ์šฉ์œผ๋กœ ๋งŒ๋“ค์–ด ์‚ฌ์šฉ๋  ์ˆ˜ ์žˆ๋‹ค. ์ƒ๊ธฐ ์กฐ์„ฑ๋ฌผ์˜ ํˆฌ์—ฌ๋Ÿ‰์€ ํ•ด๋‹น ๋ฒ•๋ฅ ์—์„œ ์ •ํ•˜๊ณ  ์žˆ๋Š” ๊ธฐ์ค€(์šฉ๋„, ์‚ฌ์šฉ๋ชฉ์ , ํ™˜์ž์˜ ์ƒํƒœ, ๋‚˜์ด, ์„ฑ๋ณ„, ์ฒด์ค‘, ์ฒ˜๋ฐฉ์•ฝ, ์งˆ๋ณ‘)์— ๋”ฐ๋ผ ์กฐ์ ˆํ•ด์•ผ ํ•œ๋‹ค.The present invention may include one or more selected from the group consisting of excipients (disintegrants, binders, lubricants), stabilizers, emulsifiers, additives, etc., which are acceptable under the pharmacological, health functional food method, food hygiene method, in addition to the extract of germinated fermented soybean. Can be. In addition, the composition may be made and used for oral use such as warnings, granules, powders, tablets, or capsules. The dosage of the composition should be adjusted according to the criteria set forth in the relevant laws (use, purpose of use, patient's condition, age, sex, weight, prescription drug, disease).

<์ ์šฉ์˜ˆ 1>; ์ œ์ œ <Application Example 1>; Formulation

์ œ์ œ๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์— ์•ฝ์‚ฌ๋ฒ•์—์„œ ํ—ˆ์šฉํ•˜๋Š” ๋ถ€ํ˜•์ œ ๋˜๋Š” ๋‹ด์ฒด๋ฅผ ์„ž์–ด ์ œ์ œ๋กœ์„œ ์‚ฐ์ œ, ์ •์ œ, ์บก์Š์ œ, ํ™˜, ๊ณผ๋ฆฝ์„ ์ œ์กฐํ•˜์˜€๋‹ค. The formulation was prepared by mixing the germinated fermented soybean extract with excipients or carriers acceptable in the pharmacological method to prepare powders, tablets, capsules, pills, and granules as preparations.

<์‚ฐ์ œ> <Product>

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 2 g ๋ฐ ์œ ๋‹น 1 g์„ ํ˜ผํ•ฉํ•˜๊ณ  ๊ธฐ๋ฐ€ํฌ์— ์ถฉ์ง„ํ•˜์—ฌ ์‚ฐ์ œ๋ฅผ ์ œ์กฐํ•˜์˜€๋‹ค. 2 g of germinated fermented soybean extract and 1 g of lactose were mixed and filled into an airtight cloth to prepare a powder.

<์ •์ œ> <Tablet>

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 100 ใŽŽ, ์˜ฅ์ˆ˜์ˆ˜์ „๋ถ„ 100 ใŽŽ, ์œ  ๋‹น 100 ใŽŽ ๋ฐ ์Šคํ…Œ์•„๋ฆฐ์‚ฐ ๋งˆ๊ทธ๋„ค 2 ใŽŽ์„ ํ˜ผํ•ฉํ•œ ํ›„, ํƒ€์ •ํ•˜์—ฌ ์ •์ œ๋ฅผ ์ œ์กฐํ•˜์˜€๋‹ค. Germinated fermented soybean extract 100 mg, corn starch 100 mg, lactose 100 mg and stearic acid magnesium 2 mg were mixed and then compressed into tablets.

<์บก์Š์ œ> <Capsules>

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 100 ใŽŽ, ์˜ฅ์ˆ˜์ˆ˜์ „๋ถ„ 100 ใŽŽ, ์œ  ๋‹น 100 ใŽŽ, ์Šคํ…Œ์•„๋ฆฐ์‚ฐ ๋งˆ๊ทธ๋„ค์Š˜ 2 ใŽŽ์„ ํ˜ผํ•ฉํ•œ ํ›„, ์ ค๋ผํ‹ด ์บก์Š์— ์ถฉ์ „ํ•˜์—ฌ ์ œ์กฐํ•˜์˜€๋‹ค.Germinated fermented soybean extract 100 mg, corn starch 100 mg, lactose 100 mg, magnesium stearate 2 mg was mixed, and then prepared by filling in gelatin capsules.

<ํ™˜> <Hwan>

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 1 g, ์œ ๋‹น 1.5 g, ๊ธ€๋ฆฌ์„ธ๋ฆฐ 1 g, ์ž์ผ๋ฆฌํ†จ 0.5 g์„ ํ˜ผํ•ฉํ•œ ํ›„, 4 g ํฌ๊ธฐ๋กœ ํ™˜์„ ์ œ์กฐํ•˜์˜€๋‹ค.After germination of fermented soybean extract 1 g, lactose 1.5 g, glycerin 1 g, and xylitol 0.5 g, a ring was prepared in a size of 4 g.

<๊ณผ๋ฆฝ><Granule>

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 150 ใŽŽ, ๋Œ€๋‘์ถ”์ถœ๋ฌผ 50 ใŽŽ, ํฌ๋„๋‹น 200 ใŽŽ, ์ „๋ถ„ 600 ใŽŽ์„ ํ”„๋ฆฌ๋ฏน์Šคํ•œ ํ›„, 30% ์—ํƒ„์˜ฌ์šฉ์•ก 100 ใŽŽ์„ ์ฒจ๊ฐ€ํ•˜์—ฌ 60 โ„ƒ์—์„œ ๊ฑด์กฐํ•˜์—ฌ ๊ณผ๋ฆฝ์„ ํ˜•์„ฑํ•œ ํ›„ ํฌ์— ์ถฉ์ง„ํ•˜์˜€๋‹ค.After germination of fermented soybean extract 150 mg, soy extract 50 mg, glucose 200 mg, and starch 600 mg, 100 mg of 30% ethanol solution was added and dried at 60 ยฐ C. to form granules, which were then filled into fabric.

<์ ์šฉ์˜ˆ 2>; ์‹ํ’ˆ<Application Example 2>; food

์‹ํ’ˆ์€ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์— ์‹ํ’ˆ์œ„์ƒ๋ฒ• ๋˜๋Š” ๊ฑด๊ฐ•๊ธฐ๋Šฅ์‹ํ’ˆ๋ฒ•์—์„œ ํ—ˆ์šฉํ•˜๋Š” ๋ถ€ํ˜•์ œ ๋˜๋Š” ๋‹ด์ฒด๋ฅผ ์„ž์–ด ๊ฑด๊ฐ•์‹ํ’ˆ ๋ฐ ์Œ๋ฃŒ๋ฅผ ์ œ์กฐํ•˜์˜€๋‹ค.Foods were prepared by mixing the germinated fermented soybean extract with excipients or carriers allowed by the food hygiene or dietary supplement method.

<๊ฑด๊ฐ•์‹ํ’ˆ> <Health food>

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 1g, ์ดˆ์‚ฐ๋น„ํƒ€๋ฏผ A 70 ใŽ, ๋น„ํƒ€๋ฏผ E 1.0 ใŽŽ, ๋น„ํƒ€๋ฏผ 0.13 ใŽŽ, ๋น„ํƒ€๋ฏผ B2 0.15 ใŽŽ, ๋น„ํƒ€๋ฏผ B6 0.5 ใŽŽ, ๋น„ํƒ€๋ฏผ B12 0.2 ใŽ, ๋น„ํƒ€๋ฏผ C 10 ใŽŽ, ๋น„์˜คํ‹ด 10 ใŽ, ๋‹ˆ์ฝ”ํ‹ด์‚ฐ์•„๋ฏธ๋“œ 1.7 ใŽŽ, ์—ฝ์‚ฐ 50 ใŽ, ํŒํ† ํ…์‚ฐ ์นผ์Š˜ 0.5 ใŽŽ, ๋ฌด๊ธฐ์งˆ ํ˜ผํ•ฉ๋ฌผ ์ ๋Ÿ‰, ํ™ฉ์‚ฐ์ œ1์ฒ  1.75 ใŽŽ, ์‚ฐํ™”์•„์—ฐ 0.82 ใŽŽ, ํƒ„์‚ฐ๋งˆ๊ทธ๋„ค์Š˜ 25.3 ใŽŽ, ์ œ1์ธ์‚ฐ์นผ๋ฅจ 15 ใŽŽ, ์ œ2์ธ์‚ฐ์นผ์Š˜ 55 ใŽŽ, ๊ตฌ์—ฐ์‚ฐ์นผ๋ฅจ 90 ใŽŽ, ํƒ„์‚ฐ์นผ์Š˜ 100 ใŽŽ, ์—ผํ™”๋งˆ๊ทธ๋„ค์Š˜ 24.8 ใŽŽ๋ฅผ ์„ž์–ด ๊ฑด๊ฐ•์‹ํ’ˆ์„ ์ œ์กฐํ•˜์˜€๋‹ค. Germinated fermented soybean extract 1g, Vitamin A 70 ฮผg, Vitamin E 1.0 mg, Vitamin 0.13 mg, Vitamin B2 0.15 mg, Vitamin B6 0.5 mg, Vitamin B12 0.2 ฮผg, Vitamin C 10 mg, Biotin 10 ฮผg, Nicotinamide 1.7 mg, Folic acid 50 ใŽ, calcium pantothenate 0.5 mg, mineral mixture appropriate amount, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monophosphate 15 mg, calcium diphosphate 55 mg, potassium citrate 90 mg, carbonate Health food was prepared by mixing 100 mg of calcium and 24.8 mg of magnesium chloride.

์ƒ๊ธฐ์—์„œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์— ๋น„ํƒ€๋ฏผ ๋ฐ ๋ฏธ๋„ค๋ž„์„ ํ˜ผํ•ฉํ•œ ๋น„์œจ์€ ์ œํ˜•์ด๋‚˜ ์ˆ˜์š”์ž์ธต์— ๋”ฐ๋ผ ๋‹ค์–‘ํ•˜๊ฒŒ ๋ณ€ํ˜•ํ•  ์ˆ˜ ์žˆ๋‹ค. The proportion of the vitamin and minerals mixed with the germinated fermented soybean extract may be variously modified according to the formulation or the consumer layer.

<์Œ๋ฃŒ> <Beverage>

๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ 1%, ์•ก์ƒ๊ณผ๋‹น 0.5%, ์˜ฌ๋ฆฌ๊ณ ๋‹น 2%, ์„คํƒ• 2%, ์‹์—ผ 0.5% ๋ฐ ์ž”๋ถ€์˜ ์ •์ œ์ˆ˜์™€ ๋ถ€์žฌ๋ฃŒ๋ฅผ ๋ฐฐํ•ฉํ•˜์—ฌ ์‚ด๊ท ํ•œ ํ›„ ํฌ์žฅํ•˜์—ฌ ์Œ๋ฃŒ๋ฅผ ์ œ์กฐํ•˜์˜€๋‹ค.Germinated fermented soybean extract 1%, liquid fructose 0.5%, oligosaccharide 2%, sugar 2%, salt 0.5% and the balance of the purified water and subsidiary materials were combined and sterilized and then packaged to prepare a beverage.

์ƒ๊ธฐ์—์„œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์— ๋‹น๋ฅ˜, ์‹์ด์„ฌ์œ , ๋น„ํƒ€๋ฏผ ๋ฐ ๋ฏธ๋„ค๋ž„์„ ํ˜ผํ•ฉํ•œ ๋น„์œจ์€ ์ œํ˜•์ด๋‚˜ ์ˆ˜์š”์ž์ธต์— ๋”ฐ๋ผ ๋‹ค์–‘ํ•˜๊ฒŒ ๋ณ€ํ˜•ํ•  ์ˆ˜ ์žˆ๋‹ค. The ratio of sugars, dietary fiber, vitamins and minerals to the germinated fermented soybean extract may be variously modified depending on the dosage form or consumer layer.

๋ณธ ๋ฐœ๋ช…์˜ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์€ ํ˜ˆ์ค‘ ์—์ŠคํŠธ๋กœ๊ฒ๊ณผ ํ…Œ์Šคํ† ์Šคํ…Œ๋ก  ๋†๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ•˜๊ณ , ์ด๋กœ ์ธํ•ด ๊ฐฑ๋…„๊ธฐ ์ฆ์ƒ์„ ์™„ํ™”์‹œํ‚ค๊ณ , ๊ณจ์˜ ๋ฏธ๋„ค๋ž„ ํ•จ๋Ÿ‰์„ ์ฆ๊ฐ€์‹œ์ผœ ๊ณจ์˜ ๋ฐ€๋„๊ฐ€ ์œ ์˜์ ์œผ๋กœ ์ฆ๊ฐ€ํ•˜๊ณ , ํ•ด๋ฉด๊ณจ์˜ ๋‘๊ป˜์™€ ์ˆ˜๊ฐ€ ์ฆ๊ฐ€ํ•œ๋‹ค. ๋”ฐ๋ผ์„œ ์‚ฐ์—…์ƒ ์ด์šฉ๊ฐ€๋Šฅ์„ฑ์ด ์žˆ๋‹ค.Germinated fermented soybean extract of the present invention significantly increases blood estrogen and testosterone concentrations, thereby alleviating menopausal symptoms, increasing bone mineral content, significantly increasing bone density, and the thickness and number of sponges Increases. Therefore, there is industrial applicability.

Claims (10)

์ฟ ๋ฉ”์Šคํ…Œ๋กค์„ ํ•จ์œ ํ•˜๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ. Germinated fermented soybean extract containing cumesterol. ์ œ1ํ•ญ์— ์žˆ์–ด์„œ, ์ƒ๊ธฐ ์ถ”์ถœ๋ฌผ์€ ๊ฐ„์„ธํฌ ๋ณดํ˜ธ์šฉ์ธ ๊ฒƒ์„ ํŠน์ง•์œผ๋กœ ํ•˜๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ. The germinated fermented soybean extract of claim 1, wherein the extract is for protecting hepatocytes. ์ œ1ํ•ญ์˜ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ํฌํ•จํ•˜๋Š” ๊ฐ„์„ธํฌ ๋ณดํ˜ธ์šฉ ์กฐ์„ฑ๋ฌผ. A composition for protecting hepatocytes comprising the germinated fermented soybean extract of claim 1. ์ œ1ํ•ญ์— ์žˆ์–ด์„œ, ์ƒ๊ธฐ ์ถ”์ถœ๋ฌผ์€ ๊ณจ ๋ฐ€๋„ ์ฆ๊ฐ€์šฉ์ธ ๊ฒƒ์„ ํŠน์ง•์œผ๋กœ ํ•˜๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ. The germinated fermented soybean extract according to claim 1, wherein the extract is for increasing bone density. ์ œ1ํ•ญ์˜ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ํฌํ•จํ•˜๋Š” ๊ณจ ๋ฐ€๋„ ์ฆ๊ฐ€์šฉ ์กฐ์„ฑ๋ฌผ. Composition for increasing bone density comprising the germinated fermented soybean extract of claim 1. ์ œ3ํ•ญ ๋˜๋Š” ์ œ5ํ•ญ์— ์žˆ์–ด์„œ, ์•ฝ์‚ฌ๋ฒ•์—์„œ ํ—ˆ์šฉํ•˜๋Š” ๋ถ€ํ˜•์ œ๋ฅผ ๋” ํฌํ•จํ•˜๋Š” ์•ฝํ•™ ์กฐ์„ฑ๋ฌผ์ธ ๊ฒƒ์„ ํŠน์ง•์œผ๋กœ ํ•˜๋Š” ์กฐ์„ฑ๋ฌผ.The composition according to claim 3 or 5, which is a pharmaceutical composition further comprising an excipient acceptable in the pharmacological method. ์ œ3ํ•ญ ๋˜๋Š” ์ œ5ํ•ญ์— ์žˆ์–ด์„œ, ๊ฑด๊ฐ•๊ธฐ๋Šฅ์‹ํ’ˆ๋ฒ•์—์„œ ํ—ˆ์šฉํ•˜๋Š” ๋ถ€ํ˜•์ œ๋ฅผ ๋” ํฌํ•จํ•˜๋Š” ๊ฑด๊ฐ•๊ธฐ๋Šฅ์‹ํ’ˆ ์กฐ์„ฑ๋ฌผ์ธ ๊ฒƒ์„ ํŠน์ง•์œผ๋กœ ํ•˜๋Š” ์กฐ์„ฑ๋ฌผ.The nutraceutical composition according to claim 3 or 5, further comprising an excipient which is allowed by the nutraceutical method. ์ œ3ํ•ญ ๋˜๋Š” ์ œ5ํ•ญ์— ์žˆ์–ด์„œ, ์‹ํ’ˆ์œ„์ƒ๋ฒ•์—์„œ ํ—ˆ์šฉํ•˜๋Š” ๋ถ€ํ˜•์ œ๋ฅผ ๋” ํฌํ•จํ•˜๋Š” ์‹ํ’ˆ ์กฐ์„ฑ๋ฌผ์ธ ๊ฒƒ์„ ํŠน์ง•์œผ๋กœ ํ•˜๋Š” ์กฐ์„ฑ๋ฌผ.The composition according to claim 3 or 5, which is a food composition further comprising an excipient acceptable in the food hygiene method. ์ œ3ํ•ญ ๋‚ด์ง€ ์ œ5ํ•ญ ์ค‘ ์–ด๋Š ํ•œ ํ•ญ์— ์žˆ์–ด์„œ, ์ƒ๊ธฐ ์กฐ์„ฑ๋ฌผ์˜ ์ œํ˜•์€ ์•ก์ œ, ๊ณผ๋ฆฝ์ œ, ํ™˜์ œ ๋˜๋Š” ์บก์Š์ธ ๊ฒƒ์„ ํŠน์ง•์œผ๋กœ ํ•˜๋Š” ์กฐ์„ฑ๋ฌผ.The composition of any one of claims 3 to 5, wherein the formulation of the composition is a liquid, granule, pill or capsule. ๋Œ€๋‘๋ฅผ ๋ฐœ์•„์‹œ์ผœ ํ™ฉ๊ตญ๊ท ์„ ์ ‘์ข…ํ•˜์—ฌ ๋ฐœํšจ์‹œํ‚ค๊ณ , ์—ฌ๊ณผํ•œ ํ›„ ๊ฑด์กฐํ•˜์—ฌ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์„ ์–ป๋Š” ๋‹จ๊ณ„;Germinating soybeans to inoculate and ferment the Rheum bacteria, and filtering and drying to obtain germinated fermented soybeans; ์ƒ๊ธฐ ๊ฑด์กฐ๋œ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ์— ์—ํƒ„์˜ฌ ์šฉ์•ก์„ ๊ฐ€ํ•˜์—ฌ ๋ถ„ํ•ดํ•œ ํ›„, ์ดˆ์ŒํŒŒ๋กœ ์—ด์ˆ˜ ์ถ”์ถœํ•˜์—ฌ ์—ด์ˆ˜ ์ถ”์ถœ๋ฌผ์„ ์–ป๋Š” ๋‹จ๊ณ„; ๋ฐDecomposing by adding ethanol solution to the dried germinated fermented soybeans, and extracting hot water by ultrasonication to obtain a hot water extract; And ์ƒ๊ธฐ ์—ด์ˆ˜ ์ถ”์ถœ๋ฌผ์„ ์›์‹ฌ๋ถ„๋ฆฌํ•˜์—ฌ ์–ป์€ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์—ฌ๊ณผํ•œ ํ›„, ๊ฑด์กฐํ•˜์—ฌ ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์„ ์–ป๋Š” ๋‹จ๊ณ„๋ฅผ ํฌํ•จํ•˜๋Š” ๊ฒƒ์„ ํŠน์ง•์œผ๋กœ ํ•˜๋Š” ๋ฐœ์•„ ๋ฐœํšจ์ฝฉ ์ถ”์ถœ๋ฌผ์˜ ์ œ์กฐ๋ฐฉ๋ฒ•.Filtrating the germinated fermented soybean extract obtained by centrifuging the hot water extract, and then drying to obtain a germinated fermented soybean extract.
PCT/KR2016/012399 2015-10-30 2016-10-31 Coumestrol-containing germinated fermented soybean extract having hepatocyte protective effect and bone density increase effect and composition containing same Ceased WO2017074159A2 (en)

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