KR20190081582A - Fermented product having an anti-obesity effect containing apigenin or naringenin as an active ingredient and a method for producing the same - Google Patents

Abstract

The present invention relates to an anti-obesity pharmaceutical composition or health food composition containing apigenin or naringenin as an active ingredient in multiple vegetable-fermented products and flavonoid-based compounds separated therefrom. According to the present invention, the vegetable-fermented products including a variety of vegetables such as broccoli, bok choy, okra, water spinach, eggplant, wax gourd, cabbage and apigenin or naringenin isolated therefrom show an excellent effect in preventing obesity by inhibiting fat accumulation by increasing energy consumption through expression of a heat-generating protein called UCP1 during intestinal vegetable degradation. Therefore, the composition containing the same may be useful as an anti-obesity pharmaceutical composition or health food composition.

Description

     A fermented product having an anti-obesity effect containing apigenin or naringenin as an active ingredient, and a fermented product having an anti-obesity effect containing apigenin or naringenin as an active ingredient and a method for producing same,

The present invention relates to a fermented product of various vegetables having an anti-obesity effect containing apigenin or naringenin as an active ingredient among flavonoid-based compounds and a process for producing the same.

     Obesity has become one of the serious diseases as mankind has developed into a prosperous society, and the World Health Organization (WHO) has declared that obesity is the subject of diseases to be treated. Obesity is a metabolic disorder caused by an imbalance in the intake and consumption of calories, and is defined as an increased state of adipose tissue. Obesity affects all age groups, especially among the elderly. The fact that obesity can cause serious diseases such as hyperlipemia, diabetes, cholelithiasis, atherosclerosis and cardiovascular diseases such as hypertension is considered to be a disease not only in the metabolic diseases or diseases but also as a disease. The risk of adult disease is 3 to 6 times higher than normal people.

The major cause of obesity has been the efforts to prevent the intake of energy from being excessively excessive as a result of excessive food supply due to the inability to control the food intake by itself. Some obese patients tried to control the amount of energy intake through dieting and exercise therapy, and other behavioral therapies, psychotherapy, pharmacotherapy, or surgical therapy were used in combination. However, due to serious side effects, It was limited.

In the case of dietary therapy, it is difficult to maintain a healthy body for a long period of time due to the lack of nutrients due to selective intake of foods having low fat or sugar content, and the psychological burden due to the fasting or control of food is increased. Improvement in metabolic rate during rest, correction of insulin resistance, and reduction of body fat. However, it is necessary to perform aerobic exercise for 20 minutes at least three times a week. However, exercise therapy usually requires unnecessary efforts, burdens, discomfort, and patience, and there are many cases where weight loss can not be continued for a long period of time. Or urgent treatment for extreme obesity is required, drug therapy or surgical therapy is performed, but surgical treatment is very burdensome to the patient. There are various kinds of drugs used for pharmacotherapy such as digestion inhibitor, obesity accumulation inhibitor, or metabolic accelerator. However, because of the use of these drugs, they may cause digestive disorders due to the burden on the human body and may cause short- There is a problem that they can not be used continuously for a long period of time. For example, there is a method of using the water-soluble substance such as cellulose and the like to inflate in the gastrointestinal tract after ingestion with water to inhibit the ingestion of additional food. However, this method causes ulcers in the gastrointestinal wall due to contact with the gastrointestinal tract for a long time, Due to the high adsorption of inorganic salts, it inhibited the absorption of inorganic salts and caused a decrease in inorganic salts. By taking natural products containing a large amount of caffeine and ephedrine, it increased the activity of autonomic nervous system and increased energy metabolism, In addition to general side effects such as gastric ulcers, induction may have side effects such as blood pressure, hypertension, palpitations, and stroke. In addition, when the fat absorption blocker is used, it has an effect of blocking the absorption of fat in the digestive organs, but it is possible to use only a very limited use because of various side effects such as diarrhea, And side effects related to the circulatory system and side effects at the same time when taking other medicines. Thus, there is a problem that it can not be used without the doctor's prescription.

As described above, various attempts have been made to treat obesity. However, until now, there has been almost no finding of a treatment method that can relieve the stability, efficacy and psychological burden.

As a result of the development of anti-obesity materials using natural materials, there are dietary fibers such as hydroxycitric acid (HCA), olive leaf, chicory, inulin, Chitosan and flavonoids are known as substances that inhibit absorption. Capsaicin of red pepper, catechin of green tea, retinoic acid and the like, and L-carnitine, conjugated linoleic acid, milk calcium and related proteins which regulate lipid metabolism have been reported as substances which induce heat generation and inhibit fat accumulation. As the herb medicine, it was reported that Zyseol, Yangyang, Kyeongjae, Green tea, pine needles, chrysanthemum, Chenguan,

Flavonoids are polyphenolic compounds with inherent properties that are involved in the color and flavor of plants and are known to protect against attack by pathogens and insects. In addition, flavonoids are representative antioxidants and exhibit anticancer, antiviral, anti-inflammatory, anti-allergic activity and are attracting attention as a substance that has a beneficial effect on human and animal health.

Oh Pinero Jean (apigenin) is the large amount as a kind of flavonoid ingredients contained in various fruits and vegetables, apples, beans, broccoli, celery, cherries, grapes, onions, parsley, tomatoes, etc., the formula is C ₁₅ H ₁₀ O ₅ , the molecular weight is 270.23, insoluble and has the effect of inhibiting inflammation and oxidative stress and promoting carbohydrate metabolism.

Naringenin is a flavonoid flavanone nonpolar compound with two benzene rings. It is a yellow colored powder with the formula C ₁₅ H ₁₂ O ₅ , a molecular weight of 272.257, a melting point of 247 to 250 / RTI > Citrus fruits such as grapes, oranges and citrus fruits. It helps to repair damaged DNA in cancer cells and has the effect of inhibiting aflatoxin B1 activity, a carcinogen. In addition, it has a very high antioxidative effect, and is known to be effective for anti-inflammation, fat reduction, promotion of carbohydrate metabolism, and control of the immune system.

Although research on the anti-obesity effect of flavonoid apigenin or naringenin has not been done much recently, it has recently been shown that when a normal diet is fed to a rats that induces obesity in a high fat diet and then fed to a high fat diet, Compared with the group in which the diet was continuously consumed, there was a high yo-yo phenomenon during the same period. The concentration of flavonoid which promotes the energy consumption of the intestine of the yo-yo was low and the expression of flavonoid-degrading enzyme was high . In obese rats, the share of bacteria that break down flavonoids, which increase energy consumption, is increased, and the causal relationship is confirmed by the fact that the intestinal flora changed in obesity lowered flavonoid. The flavonoids identified in this study were found to decompose vegetables into two classes of naringin and apigenin, and it was confirmed that the expression of heat-producing protein, UCP1, increased energy consumption and inhibited fat accumulation (Nature 20796, 2016, Persistent microbiome alterations modulate the rate of post-dieting weight regain).

Therefore, consumers are more interested in anti-obesity agents based on natural materials with less side effects, and development of safe and effective natural food materials is required to meet these demands.

It is an object of the present invention to provide a method for detecting an active ingredient having an anti-obesity effect from natural materials, wherein the flavonoid apigenin or naringenin inhibits fat accumulation by increasing energy consumption through expression of heat- It is characterized by containing an active ingredient of vegetable fermented product including a variety of vegetables such as broccoli, green tea, okra, air core, branches, Donga, cabbage and the like and apigenin or naringenin isolated therefrom By weight of the composition. The present invention also relates to a pharmaceutical composition for the prevention and treatment of obesity comprising, as an active ingredient, 33 kinds of fermented products of various kinds of vegetables such as broccoli, blueberry, okra, air core, branches, Donga and cabbage or apigenin or naringenin isolated therefrom Or a health food for obesity improvement.

In order to solve the above-mentioned problems, a method for producing a fermented product of various vegetables containing apigenin or naringenin as an active ingredient according to the present invention is as follows. The method for preparing a fermented vegetable according to the present invention is characterized in that it comprises the steps of: washing the 33 kinds of vegetables and then removing the water; preparing a material; cutting the prepared material to a predetermined size; mixing organic aloe juice and organic fructooligosaccharide Mixing step; Adding the plant lactic acid bacteria Lactobacillus plantarum PMO 08 to the material mixing step for 3 to 5 days at 37 ° C: adding the fermented material after the fermentation step to the juicer and adding the juice: adding 50% : 50% (v / v), and aging at 25 캜 at room temperature: quantitatively analyzing the aging solution with an apigenin or naringenin component by HPLC (high performance liquid chromatography) .

At this time, the 33 kinds of vegetables such as Okra, Kangdu, Yeoju, Dong-a, and Choshi are preferably grown in organic farming.

As another aspect of the present invention, a fermented product of a variety of vegetables is characterized in that it is produced by the above production method.

The 33 kinds of fermented vegetables of various kinds such as broccoli, blueberry, okra, air core, branches, Donga and cabbage contain a large amount of apigenin or naringenin as an active ingredient compared to a single vegetable or a mixture of multiple vegetables, . In addition, the fermented product of various vegetables according to the present invention has an advantage of antioxidative effect by lactic acid fermentation.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a view showing an entire process of a method for producing a fermented vegetable matter according to the present invention
FIG. 2 shows experimental results of apigenin and naringenin components of the multi-type fermented vegetable according to the present invention.
FIG. 3 is a graph showing an antioxidative effect of a fermented product of various vegetables according to the present invention.

Hereinafter, the present invention will be described in detail.

Fig. 1 shows the entire process of a method for producing a fermented vegetable matter according to the present invention. In preparing the ingredients, the main ingredients of the various vegetable fermented products are organic broccoli, ginger root, eggplant, carrot, yuzu, Donga, cauliflower, red cabbage, cabbage, lettuce, okra, eastern, asparagus, pumpkin, cucumber, Chinese green pepper It is a step to remove the water after washing the drip tomatoes, spinach, cucumbers, leeks, celery, radishes, tomatoes, potatoes, coriander leaves, corn, watermelon, Chinese cabbage, radishes, loofahs and young cucumbers.

Organic Aloe Juice: Organic Fructo-oligosaccharide = 50% Organic Fructo-oligosaccharides: Organic Fructo-oligosaccharides: Organic Fructo-oligosaccharides: Organic Fructo-oligosaccharides: : 30%: 20% (v / v).

In the fermentation step, the lactic acid bacteria Lactobacillus plantarum PMO 08 is added to the mixture, stirred and mixed at 37 ° C for 3 to 5 days, and the lactic acid bacteria is preferably added at a rate of 0.01 to 0.1%.

In the juice stage, the fermentation broth is added to the juicer and the broth and juice solution are separated to recover the juice solution.

In the aging step, the juice was mixed with 50% (v / v) 50% (v / v) of organic fructooligosaccharide and stored in a 1L to 2L container at 25 캜 for 6 months. And give a soft taste.

In the analysis step of apigenin or naringenin, 1 mL of methanol was added to 1 mL of the aged solution, and the mixture was centrifuged at 4,000 rpm for 10 minutes to obtain a supernatant, which was then filtered through a 0.45 μm filter and quantitatively analyzed by HPLC (high performance liquid chromatography) Or naringenin content of the sample.

The vegetable fermentation composition is prepared by adjusting the blending ratio of the vegetables in the descending order of apigenin or naringenin content.

   Organic broccoli, green broccoli, eggplant, eggplant, carrot, yujoo, Donga, cauliflower, red cabbage, cabbage, lettuce, okra, eastern, asparagus, pumpkin, cucumber, cabbage, green pepper, chili pepper, red tomato, spinach, , 500g each of them were prepared by crushing them in a grinder, and then 300g of organic aloe juice and 200g of organic fructo-oligosaccharide were mixed. Then, Lactobacillus plantarum PMO 08 was added at a rate of 0.01 to 0.1%, mixed with stirring, and fermented at 37 ° C for 3 to 5 days. Then, the fermentation broth was placed in a juicer and the juice was separated. 500 g of the juice was mixed with 500 g of organic fructooligosaccharide, and the mixture was placed in 1 L of airtight container to produce 33 kinds of vegetable fermentation broth.

On the other hand, the vegetables used in Example 1 were used in the same manner, and the vegetables were cut and crushed. Then, the vegetables were dipped in organic fructooligosaccharides without the fermentation step, Was used as a comparative example.

In Example 3, the flavonoid apigenin or naringenin content analysis was performed on the anti-obesity effect of the vegetable fermentation broth or the general fermented milk prepared in Examples 1 to 3 above.

1 mL of methanol was added to 1 mL of the fermented fermentation broth or ordinary crude liquid, and the mixture was centrifuged at 4,000 rpm for 10 minutes to remove the supernatant. The supernatant was filtered through a 0.45-μm filter and analyzed by HPLC (high performance liquid chromatography) The contents were quantitatively analyzed. The standard materials apigenin and naringenin reagent were purchased from Sigma and the HPLC analysis conditions are shown in Table 1 below.

The results of the apigenin or naringenin contents of the vegetable fermented or vegetable composition quantitatively analyzed by HPLC in Example 3 are shown in Table 2 below. The 33 kinds of vegetables showed higher content of apigenin or naringenin than those of ordinary vegetables, and it was confirmed that the anti-obesity effect of vegetable fermentation by lactic acid bacteria fermentation was excellent.

In Example 5, the vegetable fermentation composition of Example 4 was prepared by adjusting the percentage of vegetables in the order of higher contents based on the analysis results of apigenin or naringenin. The compounding ratios of the various vegetable fermentation compositions are shown in Table 3, and the figures showing the contents of various common vegetable compositions and apigenin or naringenin are shown in [Fig. 2].

Antioxidant activity assay ABST antioxidant efficacy experiment

The ABST antioxidant efficacy tests were conducted using the food raw materials prepared in Examples 1 to 3 and the comparative examples using the date palm extract, Manuka honey, and propolis extract, respectively. Antioxidant efficacy tests using color change were tested and modified. ABST (2,2'-azinobis- (3-ethylbenzothiazoline-6-sulfonic acid) was dissolved in 5 mM potassium phosphate buffer solution (pH 7.4) at a concentration of 7 mM, added with 2.45 mM potassium persulfate and 0.5 Immediately prior to each test, potassium phosphate buffer solution was added to each well of a 96-well plate at a rate of 200 μM, and the absorbance at 734 nm was measured using an absorbance meter to obtain an ABST reagent diluted to 1.0 To each well of the 96-well plate, add the extract prepared in concentration, reference solution and 20 uL of the reference solution, add 180 μL of ABST solution to each well, leave at room temperature for 5 minutes, and measure the absorbance at 734 nm using a spectrophotometer The ABST radical scavenging activity was calculated as a percentage as shown in the following equation:

Radical scavenging ability (%) = 100 - (Absorption and inversion of test group / absorbance value of control group) × 100

    , And the experimental results are shown in Fig.

Claims (7)

The main ingredients of various vegetable fermented products are broccoli, blueberry, eggplant, carrot, yuju, Donga, cauliflower, red cabbage, cabbage, lettuce, okra, eastern, asparagus, pumpkin, cucumber, cabbage, green pepper, Ingredients to remove the water after washing the cucumber, Chinese cabbage, leek, celery, radish, tomato, potato, coriander, corn, watermelon, Chinese cabbage,
The above-mentioned 33 kinds of vegetables which had been washed and dried were cut to a predetermined size, respectively, followed by pulverization. Then, aloe juice and fructooligosaccharide were added to the mixture to prepare organic vegetable juice: Aloe juice: fructooligosaccharide = 50%: 30%: 20% v / v). Mixing step:
Lactobacillus plantarum PMO 08, a vegetable lactic acid bacterium, is added to the mixed solution, and the mixture is stirred and then fermented at 37 ° C for 3 to 5 days;
A step of adding the fermentation broth to an extractor and separating the broth and the juice solution to recover the juice solution;
An aging step of aging the juice solution by mixing the fructooligosaccharide;
Quantitatively analyzing the aged solution with high performance liquid chromatography (HPLC) equipment and quantitatively analyzing the content of apigenin or naringenin;
Wherein the individual fermentation microcapsules after the HPLC analysis are prepared by adjusting the blending ratio in the descending order of apigenin or naringenin content
2. The method according to claim 1, wherein the broccoli is selected from the group consisting of broccoli, sweet pepper, eggplant, carrot, ladybug, Donga, cauliflower, red cabbage, cabbage, lettuce, okra, eastern, asparagus, pumpkin, cucumber, Chinese green pepper, , A method of producing a multi-type vegetable fermentation broth comprising the step of cultivating a vegetable fermentation broth comprising the steps of:
The method according to claim 1, wherein the fructooligosaccharide and the aloe juice are cultivated in an organic manner.
The method according to claim 1, wherein the vegetable lactic acid bacterium content is 0.01 to 0.1% (v / v)
The fermented broccoli composition according to claim 1, which comprises 8% by weight of broccoli, 5% by weight of chrysanthemum root, 15% by weight of branches, 1% by weight of carrot, 1% by weight of yeast, 10% 2% by weight of cabbage, 1% by weight of lettuce, 5% by weight of young cabbage, 1% by weight of okra, 2.4% by weight of asparagus, 1% 1% by weight of red pepper, 0.02% by weight of drop tomato, 0.5% by weight of spinach, 0.5% by weight of leek, 0.2% by weight of celery, 0.1% by weight of celery, 4% , 2% by weight of corn, 1% by weight of watermelon, 1.8% by weight of samsar cabbage, 0.2% by weight of marbles, 0.2% by weight of scrubbers and 0.2% by weight of young cucumber
The fermented vegetable composition according to claim 5, which contains 0.074 mg / g of apigenin and 0.055 mg / g of naringenin
The fermented vegetable composition according to claim 5, which has an anti-obesity or antioxidative effect

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