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WO2019231076A1 - Composition pour améliorer la fonction hépatique, contenant un produit fermenté de bactéries d'acide lactique de radis noir, et procédé de préparation de produit fermenté de bactéries d'acide lactique de radis noir - Google Patents

Composition pour améliorer la fonction hépatique, contenant un produit fermenté de bactéries d'acide lactique de radis noir, et procédé de préparation de produit fermenté de bactéries d'acide lactique de radis noir Download PDF

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Publication number
WO2019231076A1
WO2019231076A1 PCT/KR2019/002164 KR2019002164W WO2019231076A1 WO 2019231076 A1 WO2019231076 A1 WO 2019231076A1 KR 2019002164 W KR2019002164 W KR 2019002164W WO 2019231076 A1 WO2019231076 A1 WO 2019231076A1
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Prior art keywords
black radish
lactobacillus
lactic acid
composition
acid bacteria
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Korean (ko)
Inventor
김현규
이학성
김성은
백신화
김기옥
양다운
홍승현
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JEJU TECHNOPARK
Kolmar BNH Co Ltd
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JEJU TECHNOPARK
Kolmar BNH Co Ltd
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    • 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
    • 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
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A23L19/01—Instant products; Powders; Flakes; Granules
    • 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
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • 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
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A23L19/10—Products from fruits or vegetables; Preparation or treatment thereof of tuberous or like starch containing root crops
    • 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065—Microorganisms
    • 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
    • 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)
    • A61K36/21—Amaranthaceae (Amaranth family), e.g. pigweed, rockwort or globe amaranth
    • 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
    • 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
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/125—Casei
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/147—Helveticus
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/169—Plantarum
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/173—Reuteri
    • 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 a liver function improving composition comprising a fermented black radish, said composition relates to a composition for improving liver function comprising a fermented black radish of the genus Lactobacillus spp.
  • the liver is one of the important organs for detoxification by synthesizing nutrients absorbed from the small intestine into metabolites or transforming them into toxic substances that are harmless to humans.
  • the liver emulsifies fat to produce bile to facilitate the absorption of fat in the small intestine, and plays a most important role in relation to metabolism because it plays a role in synthesizing the absorbed fat component back to fat. .
  • fatty tissue When abnormalities occur in the liver resulting in imbalance in fat synthesis and metabolism, fatty tissue accumulates in liver cells and causes fatty liver and cirrhosis as well as obesity and type 2 diabetes, which are a type of metabolic syndrome.
  • ROS reactive oxygen species
  • black radish is called black radish or black radish, and is a kind of radish which is the root vegetable of the cruciferous family.
  • the outside is black but the inside has white flesh and is known to have various physiological activities such as antihyperlipidemia, anti-inflammatory, antioxidant, and toxin removal.
  • black radish has an effect of improving liver function. As described above, the possibility of liver dysfunction caused by stimulating external environment and natural aging is gradually increased, and thus, black radish is used in various forms. The development of health functional foods that can be ingested is actively progressing.
  • black radish like other plants in the cruciferous family, is degraded by myrosinase to form isothiocyanate as glucosinolate in vegetables is damaged by damage to the black radish tissue.
  • Spicy taste, especially the spicy taste is particularly strong compared to other radish varieties, there is a problem that is not easy to consume.
  • the liver function improvement effect is excellent as described above, the development of health functional foods using black radish was not diversified, and was limited to being developed only to cover the spicy taste by external seasoning such as pickling. .
  • the active ingredient having an effect of improving liver function contained in the black radish is not easy to separate from the component that gives a spicy taste, the black radish extract still has a spicy taste, it was also difficult to take in the form of extracts.
  • the present invention aims to solve the problems of the prior art as described above and the technical problems that have been requested from the past.
  • An object of the present invention is to provide a composition for improving liver function, comprising a black lactic acid bacterium fermentation product, which has enhanced efficacy than black radish extract and improves sensory effect than black radish to black radish extract.
  • the antioxidant effect of the black radish is also good, it is an object to provide a composition for improving liver function comprising a black radish lactic acid bacteria fermented product having liver function improvement and antioxidant efficacy.
  • An object of the present invention is to provide a health food comprising a black radish lactic acid bacteria fermented product.
  • an object of the present invention is to provide a method for producing fermented black radish.
  • the black radish used in the present invention utilizes Jeju Dosan black radish originated from Jeju Island.
  • the composition comprises a black radish fermentation of the genus Lactobacillus spp.
  • Lactobacillus genus Lactobacillus is Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus casei ( Lactobacillus casei ), Lactobacillus helveticus ( Lactobacillus helveticus ) and Lactobacillus ruteri ( Lactobacillus reuteri ) selected from the group consisting of one or more. It may be.
  • the Lactobacillus genus lactic acid bacteria may be Lactobacillus plantarum ( Lactobacillus plantarum ).
  • the composition may be a composition for improving liver function, characterized in that to improve the liver function by inhibiting the formation of fat spheres of liver tissue.
  • the fermented black radish contained in the composition for improving liver function of the present invention may be included at a concentration of 50 to 1500 ug / ml.
  • the composition for improving liver function may be a pharmaceutical composition or a food composition.
  • the present invention also provides a health food comprising a composition for improving liver function according to the present invention.
  • the production method of fermented black radish bacteria a method for producing a fermented black radish bacteria fermented by inoculating the fermented lactobacillus ( Lactobacillus spp.) Lactic acid bacteria in black radish (black radish), grinding the black radish , Inoculated and fermented by lactobacillus lactic acid bacteria to ground black radish and powdered process to sterilize the fermented black radish and then powdered.
  • Lactobacillus spp. Lactic acid bacteria in black radish (black radish)
  • black radish black radish
  • Inoculated and fermented by lactobacillus lactic acid bacteria to ground black radish and powdered process to sterilize the fermented black radish and then powdered.
  • the grinding process may further include a sterilization process of mixing and sterilizing the purified water in the crushed black radish, the sterilization process may be performed at a temperature of 50 to 150 °C, the fermentation process is lactobacillus in the sterilized ground black radish
  • the genus lactic acid bacteria may be inoculated with 5 x 10 2 to 5 x 10 10 CFU / 100g.
  • the fermentation process may be a fermentation for 12 to 120 hours at a temperature of 20 to 50 °C, the powdering process may be to freeze-dried powdered fermented black radish.
  • composition of the present invention is effective in improving liver function since it has an effect of inhibiting fat cell formation in hepatocytes, which is one of the indicators of inhibition of liver damage.
  • composition of the present invention has a further improved liver function improvement effect by improving the absorption of the ingredients effective in improving liver function of black radish which is effective in improving liver function.
  • composition of the present invention has the effect of improving liver function more excellent than the black radish extract and the black radish nutrient source.
  • composition of the present invention has an easy effect of consumers intake as the pungent taste of the black radish extract is weakened to improve the sensory effect.
  • composition of the present invention also has an excellent antioxidant effect in addition to the liver damage inhibitory effect.
  • 1 is a graph showing the cell viability of each sample
  • 3 is a graph showing the AST value of each sample.
  • the liver breaks down the digested lipolysis component of the small intestine into fat, converts some of the absorbed carbohydrates into fat, and stores it. It also decomposes toxins absorbed from the outside or nitrogen waste produced by the human body. It is a very important organ in metabolism and detoxification because it plays a role in transforming into a harmless urea component.
  • free radicals which are known to cause aging, act on proteins, lipids, and DNA of cells to cause oxidative damage, and when free radicals act on liver cells to cause oxidative damage, liver damage becomes more rapid. .
  • liver damage In particular, in adults who are aging, natural aging is likely to cause hepatic cell damage, resulting in weakened liver function. Oxidative damage, such as excessive drinking, overeating, and frequent drinking, can damage liver cells. When triggered, liver damage progresses very quickly, increasing the likelihood of causing various diseases caused by liver dysfunction, such as fatty liver or cirrhosis.
  • the present invention utilizes black radish known to be effective in improving liver function, but when fermenting black radish, in particular, the fermented product by lactic acid bacteria not only has an effect on improving liver function or inhibits liver damage, but also has an antioxidant effect, thereby causing liver damage. It was confirmed that the inhibitory effect was more excellent, the sensory effect was improved, and the present invention was completed.
  • composition for improving liver function comprising the fermented black radish bacteria according to the present invention includes the fermented black radish of the genus Lactobacillus spp.
  • fermentation refers to a process of decomposing organic substances using microorganisms or bacteria, and in the food field, fermented foods or fermented products obtained by fermenting foods or extracts are used.
  • the black radish lactic acid bacteria fermented extract has the effect of improving the liver function of black radish, as well as the antioxidant effect of preventing oxidative aging and the inhibition of liver function. Compared to the improved effect and the synergistic effect caused by the antioxidant effect has the effect of improving the liver function more effectively.
  • the Lactobacillus genus Lactobacillus is specifically, Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus casei ( Lactobacillus casei ), Lactobacillus helveticus ( Lactobacillus helveticus ) and Lactobacillus Luteri ( Lactobacillus reuteri ) in the group consisting of. It may be one or more selected, and more specifically, it may be Lactobacillus plantarum .
  • Lactobacillus plantarum ( Lactobacillus plantarum ) is also excellent in antioxidant efficacy, compared to other lactic acid bacteria, in particular, it was confirmed that the synergistic effect on improving liver function is more excellent.
  • the effect of improving liver function in the present invention may be to improve liver function by reducing hepatic cell inflammation or inhibiting the formation of fat cells in liver tissue, preferably may be to improve the function of the liver by inhibiting the formation of fat cells. have.
  • the fermented black radish contained in the composition for improving liver function of the present invention may be included at a concentration of 50 to 1500 ug / ml. It may preferably be in a concentration of 75 to 1400 ug / ml, more preferably 90 to 1300 ug / ml, most preferably 550 to 1200 ug / ml.
  • L.plantarum fermentation is excellent in the effect of inhibiting AST in the most preferred range, it is superior in the liver function improvement effect than the fermentation by other lactic acid bacteria strains.
  • the composition for improving liver function of the present invention may be a pharmaceutical composition or a food composition, and more particularly, may be provided in the form of a health food comprising the same, and in a specific form, a nutrient, health juice, health powder, Health foods such as reproductive compositions.
  • the method for producing a fermentation of black radish lactic acid bacteria inoculated by fermenting Lactobacillus spp. Lactic acid bacteria to black radish (black radish) according to the present invention, the grinding process of grinding black radish, Lactobacillus genus to ground black radish It includes a fermentation process inoculated and fermented by lactic acid bacteria and a powdering process to sterilize fermented black radish and powder.
  • the grinding process may be crushed after the sterilized black radish, specifically, the temperature for sterilization may be a heat treatment for 5 to 25 minutes at a temperature of 70 to 120 °C.
  • the heat treatment may be performed at a temperature of 80 to 110 ° C, most preferably 90 to 100 ° C.
  • the bacteria may not be completely removed, if the heat treatment at a temperature higher than the temperature, there is a problem that the active ingredient for improving the liver function of the black radish comes out from the black radish.
  • the grinding process may further include a sterilization process of mixing and sterilizing the purified black radish with sterilized black radish. Specifically, after adding purified water to the ground black radish at a ratio of 1: 0.5 to 3 by weight, 5 to 5 at 50 to 150 ° C. It may be autoclaved for 60 minutes.
  • purified water to the black radish ground in the sterilization process in a ratio of 1: 0.7 to 2, and more preferably, 1: 0.85 to 1.5 weight ratio of purified water to the ground black radish.
  • the amount of purified water is added in excess of the above ratio, there is a problem in that the amount of raw materials added per 1 LOT in the fermentation process after the sterilization process is less, so that the fermentation process is not performed smoothly, and the amount of purified water is smaller than the ratio.
  • the fermentation process is not made smoothly due to lack of fluidity in the subsequent fermentation process.
  • the sterilization process may be preferably sterilized at a temperature of 70 to 140 °C, most preferably 90 to 130 °C. If the heat treatment at a temperature lower than the temperature is less sterilization effect, when heat treatment at a higher temperature is damaged black mud tissue, may be unsuitable for fermentation by lactic acid bacteria.
  • the fermentation process may be inoculated with 5 x 10 2 to 5 x 10 10 CFU / 100g of Lactobacillus sp.
  • 5 x 10 5 to 5 x 10 9 CFU / 100g Preferably, 5 x 10 6 to 5 x 10 8 CFU / 100g.
  • the fermentation process may be to ferment for 12 to 120 hours at a temperature of 20 to 50 °C.
  • the fermentation may be performed at a temperature of 25 to 45 ° C, most preferably, 32 to 40 ° C. If the fermentation at a lower temperature than this, the fermentation itself may not occur, even when fermentation at a higher temperature than the optimum temperature for the lactic acid bacteria fermentation may not occur.
  • Fermentation time may be a fermentation at the above temperature conditions, preferably 24 to 100 hours, most preferably 30 to 72 hours fermentation.
  • the antioxidant active ingredient In the case of fermentation for a shorter time than the time, the antioxidant active ingredient is not formed sufficiently, lactic acid bacteria fermented product does not appear to have a significant level of improvement in liver function compared to the black radish or black radish extract, when fermentation for longer than the time, Degradation of the active ingredient and the black radish components by over fermentation has a problem that the liver function improvement effect is significantly reduced.
  • Strains used in the present invention are strains that can be freely distributed through the microbial application for preservation at Korea Microbial Preservation Center (KCCM).
  • Jeju black acid black radish was washed with purified water and sterilized for 15 minutes at 95 ° C., and then ground.
  • the ground black radish and purified water were mixed at a weight ratio of 1: 1, followed by autoclaving at 121 ° C. for 15 minutes.
  • Lactobacillus plantarum (KCCM 12116), Lactobacillus casei (KCCM 12452), Lactobacillus helveticus (KCCM 40989) at the Korea Culture Center of Microorganisms (KCCM) And Lactobacillus reuteri (KCCM 40717) were used for the experiment.
  • Strains received were subcultured on sloped medium and liquid medium and used for the experiment. Specifically, seeded strains were inoculated in a slope medium and incubated at 37 ° C. for 24 hours in an incubator, and when colonies were formed, seeded in culture in 37 ° C. for 48 hours in an incubator to prepare a seed culture. For spawn culture, MRS medium (Difco, USA) was used.
  • Jejudo black radish was washed with purified water and sterilized at 95 ° C. for 15 minutes, and then ground.
  • the mixed black radish and purified water were mixed at a weight ratio of 1: 1, and then autoclaved at 121 ° C. for 15 minutes, and then cooled to room temperature.
  • the seed culture medium previously cultured in 2) was inoculated at 5 ⁇ 10 8 CFU / 100g in the sterilized ground black radish mixture. Liquid fermentation by shaking culture for 48 hours in a 37 °C incubator. After fermentation was terminated at 95 °C for 15 minutes, the fermentation product was lyophilized to powder.
  • Jejudo black radish was washed with purified water, and 15 times of purified water was added thereto, followed by primary extraction for 4 hours at 95 ° C.
  • the washed black radish was first extracted at 70 ° C. for 4 hours by adding mixed alcohol of 30%, 50%, and 70% (percent concentration) instead of 15 multiples of purified water.
  • the supernatant was used for the experiment after ultrasonic extraction for 30 minutes by adding 10% DMSO. 10% DMSO was used as a control for each experiment.
  • This test method is to measure the extent to which ABTS radical cation is removed by the sample and the color of the reagent becomes light.
  • ABTS reagent was dissolved in 3.45 mM potassium persulfate and 7 mM 2,2 ⁇ -Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS, Sigma Co., USA) in tertiary distilled water. It was left to stand in the dark for 12 hours and used.
  • L.plantarum fermentation showed high activity compared to fermented by black radish extract, nutrients and other strains at 2.83%, 11.29%, 19.87% at all experimental concentrations, 250 ug / ml, 500 ug / At ml concentration, L.plantarum fermented products showed the highest activity compared to other fermented products.
  • the total polyphenol compounds and total flavonoid compounds content as indirect indicators of antioxidant efficacy were measured.
  • the total polyphenol content was measured by mixing 100 ul of Folin-denis reagent (Sigma, USA) with 20 ul of the sample according to the Folin-denis method (9), reacting at 40 ° C for 1 minute, and then 80 ul of a 7.5% Na 2 CO 3 solution. It was added and reacted at 40 degreeC for 15 minutes. After the reaction, the absorbance was measured at 765 nm.
  • the total polyphenol content was expressed as gallic acid equivalent (GAE) by substituting the absorbance of the sample in the standard curve prepared in the same way using the gallic acid standard. The experiment was repeated three times, and Duncan analysis confirmed the significance at p ⁇ 0.05 level.
  • the total flavonoid compound content was measured according to the method of Davis (10). Diethylene glycol 200 ul and 1 N NaOH 20 ul were mixed in 20 ul of the sample, followed by reaction at 37 ° C. for 1 hour, and then the absorbance was measured at 420 nm. The total flavonoid content was expressed as naringin equivalent (NE) by substituting the absorbance of the sample before the standard grain prepared by the same method using the naringin standard in advance. The experiment was repeated three times, and Duncan analysis confirmed the significance at p ⁇ 0.05 level.
  • Total polyphenol content and total flavonoid content are shown in Table 2.
  • purified water extract showed the highest content.
  • black radish lactic acid bacteria fermented products showed higher total polyphenolic compound content than black radish alcohol extract, and the content increased significantly compared to the black radish nutrient source.
  • purified water and alcohol extract were measured below the detection limit, but the black radish lactic acid bacteria fermented products were all measured flavonoid compounds. It can be seen that the increase in the flavonoid compound through the fermentation process, among which L.plantarum fermented products showed the highest content of 0.450 mg / g.
  • the black radish lactic acid bacteria fermented product had better antioxidant effect than the black radish nutrient source or the black radish extract samples.
  • MTT reagent was used after dissolving Thizolyl blue tetrazolium bromide (Sigma, USA) in 1xPBS so that the final concentration was 5 mg / ml.
  • 1x10 5 cells / ml were dispensed into a 96-well plate and incubated for 24 hours, and each sample dissolved in the medium was treated by concentration (100 ug / ml, 500 ug / ml, 1000 ug / ml). Incubated for 24 hours. After incubation, the MTT reagent was added at a ratio of about 10: 1 and reacted for 3 hours.
  • Hep G2 cells were dispensed in a 24-well plate at 2x10 5 cells / ml and incubated for 24 hours. Each sample was treated with 100 ug / ml and 500 ug / ml concentrations, and after 1 hour, oleic acid (OA) (Sigma, USA) was treated with 600 uM concentration for 24 hours. As a positive control, milk thistle extract, an individual functional ingredient, was used at a concentration of 500 ug / ml.
  • OA oleic acid
  • OA treatment group was confirmed that about 2.21 times the fat spheres were formed compared to the untreated group.
  • 500 ug / ml of milk thistle extract used as a positive control inhibited 125.25% of fat globules and slightly higher levels of fat globules were formed.
  • L.plantarum fermented product was found to have a similar level of fat globule formation (117.77%) to the untreated group (100.00%).
  • ALT and AST tests are commonly used for diagnosing liver disease.Alt and AST elevations suggest hepatic damage.In this experiment, ALT and AST inhibition was By confirming the effect, the experiment was conducted to confirm the effect of improving liver function.
  • Hep G2 cells were dispensed in 6-well plate 2x10 5 cells / ml and incubated for 24 hours. After treating the cells with ethanol (40 ul / ml), each sample was treated at a concentration of 100 ug / ml, 500 ug / ml, 1000 ug / ml. After 24 hours, cells were detached using the buffer of each kit (abcam ⁇ , cat # ab105134 and ab105135) to measure ALT and AST activity, and then the procedure was performed according to the kit protocol.
  • milk thistle extract an individual functional ingredient, was used at a concentration of 500 ug / ml. The experiment was repeated three times, and ANOVA analysis confirmed the significance at the p ⁇ 0.05 level. ALT measurement results are shown in FIG. 2 and AST measurement results are shown in FIG. 3.
  • ALT was measured as 20.6 mU / ml in the control group (cell before ethanol treatment), and increased to 33.8 mU / ml after ethanol treatment.
  • Milk thistle extract treatment group used as a positive control group was 24.6 mU / ml significantly reduced by 27% compared to the ethanol treatment group.
  • the ALT level of the cells treated with the purified water extract and the ethanol extract at the concentrations of 100 and 500 ug / ml was measured from 27.56 mU / ml to 28.54 mU / ml, which reduced the ALT level by 15.6 to 18.5% compared to the ethanol treated group.
  • As a numerical value it was confirmed that the milk thistle extract treatment group had a level of efficacy that was less than the ALT inhibitory effect.
  • ALT level was measured from 18.66 ( L.plantrum fermented product) to 24.19 mU / ml, which was reduced by 28.5 ⁇ 44.8% compared to ethanol treated group. In comparison with the simple black radish extract treatment group, it was confirmed that the black radish lactic acid bacteria fermented product had a particularly excellent ALT inhibitory effect.
  • L. plantarum fermented products suppressed ALT by 34.0%, 37.9%, and 44.8%, respectively, according to the concentration, compared to those of the positive control milkmilk extract and other black radish lactic acid bacteria. It was confirmed that it has excellent efficacy.
  • ALT measurement of fermented water samples by type of black radish bacteria showed that L.reuteri fermentation, L.casei fermentation, and L.helveticus fermentation suppressed ALT at the same level as the milk thistle extract in all concentration samples.
  • L.plantrum fermented products it was confirmed that ALT inhibitory effect was superior to the milk thistle extract in all concentration samples.
  • the ALT inhibitory effect was nearly 50% compared to the cells after ethanol treatment, which was compared with the positive control milkmilk extract treatment group. It can be seen that the comparison has a significant ALT inhibitory effect.
  • AST was measured to be 7.57 mU / ml in control (cells before ethanol treatment) and increased to 10.88 mU / ml after ethanol treatment.
  • the milk control group a positive control group, was 8.71 mU / ml, and the AST level was reduced by 19.9% compared to the ethanol treatment group.
  • the black radish nutrient treatment group showed values of 10.53 mU / ml, 7.77 mU / ml, and 8.88 mU / ml at each concentration, and significantly decreased compared with the control group only in the 500 and 1000 ug / ml concentration treatment groups.
  • the purified water extract treatment group was measured to be 7.72 mU / ml ⁇ 9.50 mU / ml AST value of 1000 ug / ml, ethanol extract treatment group has a significant effect only in 500 ug / ml or more treatment group.
  • Simple black radish extract treatment group was confirmed to have AST inhibitory effect only when using a relatively high concentration of the sample.
  • Lactic acid bacteria fermented black radish decreased AST level significantly at all concentrations.
  • L.casei, L.helveticus and L.reuteri fermentations showed similar levels of AST inhibitory effects in all concentrations of samples regardless of concentration.
  • the L.casei fermentation treatment group was 6.92 mU / ml
  • the L.helveticus fermentation treatment group was 6.61 mU / ml
  • the L.reuteri fermentation treatment group was 6.71 mU / ml, respectively, compared to the ethanol treatment group 36.3. %, 39.2%, 38.3% AST decreased.
  • the L.plantarum fermentation had values of 6.12 mU / ml and 5.52 mU / ml in the 500 and 1000 ug / ml treatment groups, respectively, reducing AST levels to 43.8% and 49.3%, respectively. Having an AST inhibitory effect of nearly 50%, it was confirmed that the effect of improving liver function is very excellent among the black radish lactic acid bacteria fermentation products.

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Abstract

L'invention concerne une composition pour améliorer la fonction hépatique, contenant un produit fermenté de bactéries d'acide lactique d'un radis noir, la composition pour améliorer la fonction hépatique contenant un Lactobacillus spp. L'invention concerne également un produit fermenté de bactéries d'acide lactique d'un radis noir.
PCT/KR2019/002164 2018-05-31 2019-02-21 Composition pour améliorer la fonction hépatique, contenant un produit fermenté de bactéries d'acide lactique de radis noir, et procédé de préparation de produit fermenté de bactéries d'acide lactique de radis noir Ceased WO2019231076A1 (fr)

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