KR20200016610A - Composition for Improvement of Fatty Liver - Google Patents

Abstract

The present invention relates to a composition for alleviating fatty liver and, more specifically, to a food composition for alleviating fatty liver comprising, as an active ingredient, fermented products obtained by sterilizing fermented products of rice bran by lactic acid bacteria and yeast and adding the sterilized fermented products to Citrus unshiu peel, and fermenting the same in a natural state. According to the present invention, a composition for diet, a composition for alleviating fatty liver, a composition for protecting liver cells, a composition for relieving hangover, and the like can be provided.

Description

Composition for Improvement of Fatty Liver

The present invention relates to a fatty liver improver composition, and more particularly, to a fatty liver improver composition comprising a dermis extract or fermented product.

Fatty liver refers to a condition in which lipid fears, especially triglycerides, are excessively accumulated in the liver and fat fear is observed in more than half of the liver cells because normal fat metabolism is not achieved.

Clinically, fatty liver is classified as fatty liver when more than 5% of liver weight.

The causes of fatty liver include alcohol, diabetes, obesity, malnutrition (long-term low-protein diet, high-fat diet, hunger, vitamin deficiency, excess sugar intake), drug abuse (Sung Hoon Kim et al., "Cause of fatty liver diagnosed by abdominal ultrasound") Korean Journal of Medicine 175 ('95 .12) pp. 785-794).

Fatty liver is classified into cholesterol fatty liver and triglyceride fatty liver. The most common fatty liver is fatty liver when triglyceride is abnormally accumulated in liver.

Commercially available fatty liver treatments include Legaron ™ (Bugwang Pharm., Korea), Lycaba ™ (Yuhan, Korea), Heparin ™™ (Boryeong Pharm., Korea), Heparmels ™ (Hanhwa Pharmaceutical, Korea), etc. This may be exemplified, and as the prior art, Korean Patent Publication No. 2006-0028561 using Genistein, etc., and Korean Patent No. 0568550 using Rosewood Fragrance Extract may be exemplified.

On the other hand, the dermis is the skin of the citrus sunki (Jeju's native tangerine) has been reported antioxidant activity, antimicrobial activity, anti-inflammatory activity, blood pressure lowering activity (Jeong WS, Park SW, Chung SK. Food Sci. Biotechnol. 6: 292-296 (1997); Kim YC, Koh KS, Koh JS. Food Sci. Biotechnol. 10: 483-487 (2001); Cho C, Park BJ, Chung SH, Kim CB, Cha BS, Byun MW. Food Sci. Biotechnol. 13: 384-386 (2004); Murakami A, et al. Carcinogenesis 21: 1843-1850 (2000)).

An object of the present invention is to provide a fatty liver improver composition.

Other technical problems of the present invention will be presented below.

In one aspect, the present invention relates to a fatty liver improver composition comprising an dermis-derived extract or fermented product as an active ingredient.

In order to confirm fatty liver inhibitory activity as another physiological activity of the dermis, the present inventors have identified a total of five kinds of dermis-derived extracts or fermentations, that is, extracts of ethanol aqueous solution of dermis and extracts of ethanol aqueous solution of dermis, as confirmed in the following examples. A water fraction, a fermentation product using lactic acid bacteria and yeast of this water fraction, a natural fermentation product of dermis, and a natural fermentation product of dermis using rice bran fermentation product by lactic acid bacteria and yeast were prepared, and as confirmed in the following experimental example, As a result of confirming that each extract or fermented product had fatty liver inhibitory activity, all of them were confirmed to have fatty liver inhibitory activity.

The fatty liver improver composition of the present invention is provided based on the results of these experiments.

Therefore, the fatty liver improver composition of the present invention is characterized by including any one of the following dermal-derived extracts or fermented products of (I) to (IV) as an active ingredient.

(I) extracts of ethanol, water or mixed solvents of dermis and fractions thereof

(II) fermented product by lactic acid bacteria and yeast of said extract or fraction;

(III) fermented product obtained by fermenting the dermis in a natural state; And

(IV) A fermented product obtained by sterilizing and adding fermented rice bran by lactic acid bacteria and yeast to the dermis and fermenting in a natural state.

As used herein, "fatty liver" is meant to include both types of cholesterol fatty liver and triglyceride fatty liver as previously classified, and includes both types of alcoholic fatty liver and non-alcoholic fatty liver depending on the cause of fatty liver. I mean. It is shown that the following Examples and Experimental Examples show that the dermis-derived extract or fermented product has the activity of lowering the total cholesterol content and triglyceride content between the experimental animals, whether alcoholic fatty liver or non-alcoholic fatty liver. Fatty liver is due to the accumulation of cholesterol or triglycerides in the liver.

In addition, in the present specification, the "improvement" is meant to include prevention of symptoms, alleviation of symptoms and removal of symptoms.

In addition, in the present specification, the "extract fraction" means a fraction of each solvent, that is, an ethanol fraction or water fraction obtained by separating these solvents when the extraction solvent is a mixed solvent of ethanol and water, that is, an ethanol aqueous solution. Although only the fatty liver improving activity of the water fraction is disclosed in the following Examples and Experimental Examples, it is obvious that the ethanol fraction will also have the fatty liver improving activity in that the following Examples and Experimental Examples disclose that the aqueous ethanol extract extracts have improved the fatty liver. Therefore, the meaning of the fraction of the extract should be understood as including the ethanol fraction as well as the water fraction directly confirmed its activity in the following examples and experimental examples.

In addition, in the present specification, the "natural state" means a state in which no artificial manipulation is applied in temperature, humidity, CO ₂ concentration, microorganisms to be inoculated, presence of oxygen, or the like for fermentation. Therefore, fermentation in the natural state means that fermentation proceeds under aerobic conditions and at room temperature without artificially inoculating any microorganisms.

In addition, in the present specification, the "lactic acid bacteria" may be understood as bacteria that produce lactic acid by decomposing sugars such as glucose. Lactic acid bacteria were Lactobacillus sp . , Streptococcus sp . , And Pediococcus spp . sp . ), Leuconostoc sp . ) And a Bifidobacterium lactic acid bacteria (Bifidobacterium sp . The lactic acid bacteria are meant to include all of these lactic acid bacteria. However, in the following examples, Lactobacillus lactic acid bacteria were used, preferably, the lactic acid bacteria may be understood as Lactobacillus lactic acid bacteria, and in particular, may be understood as Lactobacillus plantarum ( Lactobacillus plantarum ).

Also herein, the term “yeast” refers to Saccharomyces spp. Yeast, for example Saccharomyces rouxii), saccharose in my process serenity non-zero (Sacharomyces cereviciae), access to the MY Ob pole miss Saccharomyces (Saccharomyces oviformis ), Saccharomyces steineri and the like.

In addition, in the present specification, the "rice bran" is a by-product obtained in the process of refining brown rice with white rice, and refers to including rice snow and a whistle layer. It is to be understood that the rice bran is included in the rice bran even if it additionally includes the outer skin and / or seed of the outer shell of brown rice and the endosperm and / or the endosperm, as long as it includes the rice bran and the whistle layer.

On the other hand, as shown in the following Examples and Experimental Examples as the active ingredient of the fatty liver improver composition of the present invention, the most active is a fermentation product obtained by sterilizing the fermentation of rice bran by lactic acid bacteria and yeast added to the dermis and fermentation in natural state ( IV). This fermented product is much higher in activity than fermented product (III) obtained by fermenting dermis only in natural state without adding fermented product of rice bran by lactic acid bacteria and yeast. When rice bran is fermented by lactic acid bacteria and yeast, it is speculated that a substance is newly produced which promotes the fermentation in the natural state of the dermis in a useful direction (that is, in the direction of increasing the physiological activity of the dermis).

In order to obtain the fermented product (IV), fermentation by lactic acid bacteria and yeast of rice bran should be preceded. In this case, carbon sources such as water, glucose, and sucrose are generally added to promote fermentation. Even without water, rice bran moisture or rice bran carbon source may be used, so adding water or carbon source is not necessary for fermentation by lactic acid bacteria and yeast of rice bran as long as adequate time is secured.

And before the fermentation of the rice bran is added to the dermis, the sterilization should be done, such sterilization can be done using any sterilization method known in the art, even if using a saturated salt solution in the following examples.

And the amount of the fermented rice bran is added to the dermis powder is preferably 0.5 parts by weight to 10 parts by weight based on 100 parts by weight of the dermis. If it is 0.5 parts by weight or less, the effect of promoting the fermentation of the rice bran fermentation in a useful direction will be insignificant, and if it is 10 parts by weight or more, the fermentation in the natural state is not intended by the inventors (ie, lowers the physiological activity of the dermis Direction).

Meanwhile, the fatty liver improver composition of the present invention may include any of the four dermis-derived extracts or fermented products, which are the active ingredients, in any amount depending on the use, formulation, and blending purpose, and is generally based on the total weight of the composition. 0.001% to 20% by weight.

On the other hand, fatty liver may progress to hepatitis, cirrhosis, liver cancer and the like if left untreated.

Therefore, the four dermis-derived extracts or fermented products may be useful for the purpose of preventing hepatitis, liver cirrhosis or liver cancer, which are diseases caused by fatty liver.

Therefore, in another aspect, the present invention can also be understood as a composition for the prevention of diseases caused by fatty liver comprising the extract or fermented products of the four dermis as an active ingredient.

In another aspect, the present invention relates to an obesity improving composition comprising the four dermis-derived extracts or fermented products as an active ingredient.

In the specification, "obesity" refers to a condition in which fat tissue is abnormally increased, whether it is caused by genetic factors or obesity due to environmental factors, and obesity according to the classification of body mass index (when BMI is 30.0 or more) and overweight (The meaning includes the case where the BMI is 25-30.

In the obesity improver composition of the present invention, with respect to the preferred embodiment of the active ingredient, its content and the like, the above-described bar is effective as it relates to the fatty liver improver composition of the present invention.

In another aspect, the present invention relates to a composition for a diet comprising the four dermis-derived extracts or fermented products as an active ingredient.

As used herein, "diet" is defined as a condition in which a weight condition is not obese or overweight, but for which a reduction in weight / body fat is desirable or necessary for cosmetic or health purposes. Usually the composition for a diet of the present invention will be prepared for use for beauty or health purposes of normal people.

In the composition for a diet of the present invention, in relation to the preferred embodiment of the active ingredient, its content and the like, the above-described bar is effective as it relates to the fatty liver improving composition of the present invention.

In another aspect, the present invention can be understood as a composition for protecting liver cells comprising the four dermis-derived extracts or fermented products as an active ingredient.

As confirmed in the following Examples and Experimental Examples, the extracts or fermented products derived from the four dermis were glutamic acid pyruvate transaminase (hereinafter referred to as "GPT") and glutamate oxaloacetate transaminase (hereinafter referred to as "GOT"). The four dermis-derived extracts or fermentations are useful for protecting liver cells, as they possess the activity of lowering the activity of ") and have the protective activity of HepG2, a liver cell that is damaged by alcohol. Can be.

In the above, GPT and GOT are enzymes that are used as general indicators of liver dysfunction because the liver is secreted into the blood from the liver when damaged due to obesity alcohol or the like.

Also in the composition for liver cell protection of this invention, what was described with respect to the fatty liver improver composition of this invention as mentioned above regarding the preferable aspect of its active ingredient, its content, etc. is effective as it is.

In another aspect, the present invention relates to a hangover relief composition comprising the four dermis-derived extracts or fermented products as an active ingredient.

Hangovers, such as nausea and headaches after drinking too much, are caused by aldehydes in alcohol metabolism. Generally, alcoholase (ADH) and aldehydease (ALDH) are present in the liver, and when alcohol enters the body, it is converted into acetate, which is harmless to the body.

As confirmed in the following examples, the four dermis-derived extracts or fermented products may be usefully used for hangover relief in that they promote ADH activity and ALDH activity.

In the hangover-relieving composition of the present invention, as described above with respect to the fatty liver improving composition of the present invention as described above with respect to the preferred embodiment of the active ingredient, its content and the like is effective as it is.

On the other hand, the fatty liver improving composition, the composition for preventing diseases caused by fatty liver, obesity improving composition, diet composition or hangover composition (hereinafter collectively referred to as "composition of the present invention") in a specific embodiment It can be understood as a pharmaceutical composition.

The pharmaceutical composition of the present invention may include pharmaceutically acceptable conventional ingredients in addition to the above four dermal-derived extracts or ferments, which are the active ingredients. As commonly used, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup , Methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like.

The pharmaceutical compositions of the present invention may also further comprise lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like as additives. Components suitable for each use, such as lubricants, wetting agents, etc. are already known in the art and those skilled in the art can select and use appropriate components.

Such ingredients may be included in the pharmaceutical composition of the present invention in an amount of from 85% to about 99.99% by weight, preferably from about 90% to about 99.99% by weight relative to its total weight.

On the other hand, the pharmaceutical composition of the present invention can be administered by various routes, for example, orally or rectally, or by injection into veins, muscles, subcutaneous, intrauterine dural, and the like.

The pharmaceutical compositions of the present invention may be prepared in unit dose form or may be prepared by incorporation into a multidose container. In this case, the formulation may be in the form of a solution, suspension, or emulsion, or may be in the form of elexirs, extracts, powders, granules, tablets, and the like.

The pharmaceutical composition of the present invention usually has a daily dosage in the range of 0.001 to 150 mg / kg body weight within a range that is not toxic to the human body, and may be administered once or in several divided doses. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as the route of administration, the age, sex, weight of the patient, the severity of the patient and the like, the dosage may limit the scope of the present invention in any aspect. It should not be understood as.

The composition of the present invention can be regarded as a food composition in specific embodiments.

The food composition of the present invention may be prepared from gums, vitamin complexes, dietary supplements, special nutritional supplements, functional drinks (particularly in the case of hangover relief composition of the present invention), and the like.

In addition to the four dermal-derived extracts or fermented products, the food composition of the present invention may be added with glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, or polysaccharides such as dextrin and cyclodextrin. Sugar alcohols such as xylitol, sorbitol, erythritol and the like may also be added, sweeteners such as taurine, stevia extract and the like may also be added, and other food additives may also be added.

As mentioned above, according to the present invention there is provided a fatty liver improver composition. In addition, according to the present invention there is also provided a diet composition, fatty liver improver composition, liver cell protective composition, hangover composition and the like.

It will be described below with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

<Example> Dermis Extract, Fermented Dermal Extract and Dermis Fermented Product

Example 1 Preparation of Dermis Extract

1L of 80% ethanol aqueous solution was added to 100 g of the dermis powder obtained by drying and pulverizing the dermis at room temperature, and extracted at room temperature for 7 days, followed by filtration (Whatman GF / C Fiter), and the filtered extract was concentrated under reduced pressure and lyophilized. Was prepared. This extract was stored at −20 ° C. and used in the experiments below.

Example 2 Preparation of Dermis Extract

1L of 80% ethanol aqueous solution was added to 100 g of dermis powder, extracted at room temperature for 7 days, filtered (Whatman GF / C Fiter), and the filtered extract was concentrated by a rotary concentrator to remove the supernatant ethanol fraction and freeze the remaining water fraction. Dried to prepare a dermis extract. This extract was stored at −20 ° C. and used in the experiments below.

Example 3 Preparation of Fermented Dermal Extract

The dermis extract obtained in <Example 2> was autoclaved, and the sterilized dermis extract 50% (w / w), sucrose 5% (w / w) and 5% (w / w) in a mixture of water w) inoculated with Lactobacillus plantarum (ATCC 8014) (3X10 ⁶ cells / mL) and 5% (w / w) Saccharomyces cerevises (IFO 0203) 8014) (4X10 ⁶ cells / mL) Fermented anaerobicly at 38 ° C. for 7 days and lyophilized to obtain fermented product of dermis extract. This fermentation was stored at -20 ° C and used for the experiments below.

Example 4 Preparation of Dermal Fermented Product

0.1 L of water was added to 1 kg of dermis powder and fermented in a natural state for about 2 weeks to obtain a fermented product having a water content of 15%. This fermentation was stored at -20 ° C and used for the experiments below.

Example 5 Preparation of Dermal Fermented Product

5% (w / w) Lactobacillus plantarum (ATCC 8014) (3X10 ⁶ cells / mL) and 5% (w / w) Saccharomyces cerevisiae (IFO 0203) 8014) (4X10 ⁶ ) cells / mL) and incubated anaerobicly at 38 ° C. for 7 days, and mixed the salt saturated solution in a 1:50 weight ratio (fermented solution: saturated salt solution) and sterilizing with salt and filtering (Whatman GF / C). Fiter). Next, 0.1L of water was added to 1 kg of dermis powder, and 1% (w / w) of the filtrate was added thereto, followed by natural fermentation at room temperature for about 2 weeks to obtain a fermentation product having a water content of 15%. This fermentation was stored at -20 ° C and used for the experiments below.

Experimental Example Experiments with improving obesity

Experimental Example 1 Experiment of improving obesity such as dermis extract

Three-week-old ICR male rats purchased from Daehan Biolink Co., Ltd. (Seoul, South Korea) were acclimated for one week with solid feed and water, and randomly divided rats of similar weight into three groups of 13 rats. The normal group (I) is a normal diet, the obesity control group (II) is a high fat diet, and the experimental group (III) is a high fat diet and a mixture of the extracts or fermentation products of the above examples (the high fat diet is 100 parts by weight of the extract of each example) Or 0.2 parts by weight of the fermented product was dissolved in water and mixed), and fed one by one for 97 days. The breeding environment was maintained at a temperature of 23 ± 2 ° C., a humidity of 50 ± 5%, and a night and day cycle for 12 hours.

The normal diet used Laboratory Rodent Diet 5001 (PMI Nutrition International, United States) feed, and the high-fat diet was Dyets # 101556 (> 17.7% protein, 40% fat, 31.4% carbohydrate) (Dyets Inc., United States). ) Feed was used.

Body weight was measured using an animal weight scale at the beginning of the experiment and after 97 days of breeding. The average body weights of the 39 experimental rats in three groups at the start of the experiment and the average body weight of each experimental group after breeding for 97 days are shown in Tables 1 to 7 below for each of the extracts or fermentations administered. Initial body weight was expressed as the average body weight of 39 mice in 3 groups.

When the extract of <Example 1> is administered division Initial weight (g) Final weight (g) Normal group (Ⅰ)
28.32 ± 2.13
46.86 ± 2.56 Obesity control group (Ⅱ) 52.63 ± 7.47 Experimental group (Ⅲ) 50.47 ± 5.46

* Is p <0.05 and ** is p <0.01.

When the extract of <Example 2> is administered division Initial weight Final weight Normal group (Ⅰ)
26.59 ± 3.25
45.46 ± 3.21 Obesity control group (Ⅱ) 52.57 ± 5.63 Experimental group (Ⅲ) 49.42 ± 5.76

* Is p <0.05 and ** is p <0.01.

When the fermentation product of <Example 3> is administered division Initial weight Final weight Normal group (Ⅰ)
26.45 ± 3.32
46.98 ± 2.78 Obesity control group (Ⅱ) 55.45 ± 8.28 Experimental group (Ⅲ) 51.52 ± 5.37 *

* Is p <0.05 and ** is p <0.01.

When the fermentation product of <Example 4> is administered division Initial weight Final weight Normal group (Ⅰ)
28.58 ± 2.31
47.59 ± 2.47 Obesity control group (Ⅱ) 59.27 ± 4.42 Experimental group (Ⅲ) 52.21 ± 2.31 **

* Is p <0.05 and ** is p <0.01.

When the fermentation product of <Example 5> is administered division Initial weight Final weight Normal group (Ⅰ)
28.32 ± 3.92
46.27 ± 2.43 Obesity control group (Ⅱ) 58.92 ± 3.25 Experimental group (Ⅲ) 47.82 ± 3.56 **

* Is p <0.05 and ** is p <0.01. When the extracts of <Examples 1 and 2> were fed, the weight of the experimental group was lowered as compared to the weight of the obese control group, but there was no significant difference. To 5> fermented products, the weight of the experimental group was significantly reduced compared to the weight of the obese control group. In particular, when the fermentation product of <Example 5> was administered, the weight loss effect was the highest.

Experimental Example 2 Hangover Relief Activity Experiment

Whether the extract or fermented product of each example had hangover-relieving activity was measured through alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activity.

The activity of ADH and ALDH was performed with a slight modification of the method of Bostian et al. (Bostian KA, et al. Biochem. J. 173: 787-798 (1978)).

First, the ADH fraction and the ALDH fraction were obtained from S9 rat liver homogenate (Moltox Co., Boone, Nc, USA), and the reaction mixture for the analysis of ADH activity was 1.4 ml of distilled water, 0.75 ml of 1M Tris-HCl (pH 8.8). , 0.3 ml of 20 mM NAD +, 0.3 ml of ethanol, 0.15 ml of ADH fraction, and 0.3 ml of sample, the reaction mixture for ALDH activity analysis was 2.1 ml of distilled water, 0.3 ml of 1M Tris-HCl (pH 8.8) , 0.1 ml 20 mM NAD +, 0.1 ml 3M KCl, 0.1 ml 0.33 M 2-mercaptoethanol, 0.1 ml ALDH fraction, and 0.3 ml sample.

The activity was measured for 5 minutes at 30 ° C., and then the rate of NADH production from NAD + was measured using absorbance at 340 nm. The results are shown in Table 6 below.

Relative units of ADH and ALDH

division ADH activity ALDH activity CTL 1 ± 0.18 1 ± 0.13 Extract of <Example 1> 1.5 ± 0.12 ** 1.3 ± 0.07 ** Extract of <Example 2> 1.7 ± 0.11 ** 1.3 ± 0.10 ** Fermented product of <Example 3> 2.3 ± 0.09 ** 1.8 ± 0.04 ** Fermented product of <Example 4> 2.4 ± 0.08 ** 2.0 ± 0.07 ** Fermented product of <Example 5> 3.1 ± 0.11 ** 2.7 ± 0.09 **

CTL is a non-treated group, negative control, * is p <0.05 and ** is p <0.01. It can be seen that the extract or fermentation of each of the above examples significantly increased the activity of ADH and ALDH. In particular, similar to the above results, it can be seen that the fermentation product of Example 5 significantly increased the activity of ADH and ALDH.

<Experimental Example> Protective activity experiment of hepatocyte HepG2

Hepatocellular HepG2 protective activity of the extracts or fermentations of the above examples was measured by MTT assay.

HepG2 cells were purchased from Daehan Biolink Co., Ltd. (Seoul, South Korea), and the cells were used in the experiment while culturing in a humidified incubator maintained at 5% CO ₂ using DMEM medium containing 10% FBS.

The experiment was followed by Parrizas et al. (Parrizas M, et al. J. Biol. Chem. 272: 154-161 (1997)), first inoculating cells in 24-well culture dishes and extracting or fermenting each of the above examples. Water (untreated, 0.125 mg / mL, 0.5 mg / mL and 1.0 mg / mL) was treated for 30 minutes by concentration, then treated with 5% ethanol and incubated for 48 hours. Next, the culture medium was removed and treated with 200 ml of MTT reagent (Sigma, St. Louis, MO, USA) for 1 hour at 37 ° C, and then the same amount of 2-propanol as the MTT reagent was added to lyse the cells. After inducing the reaction, the absorbance at 570 nm was measured.

The results are shown in the following <Table 7> to <Table 11>.

MTT activity when the extract of Example 1 was treated division MTT activity (A ₅₇₀ ) CTL 0.261 ± 0.052 0 mg / mL + 5% ethanol 0.062 ± 0.041 0.125 mg / mL + 5% ethanol 0.063 ± 0.021 0.5 mg / mL + 5% ethanol 0.065 ± 0.013 1 mg / mL + 5% ethanol 0.175 ± 0.021 **

CTL is untreated, * is p <0.05 and ** is p <0.01.

MTT activity when the extract of Example 2 was treated division MTT activity (A ₅₇₀ ) CTL 0.261 ± 0.052 0 mg / mL + 5% ethanol 0.062 ± 0.041 0.125 mg / mL + 5% ethanol 0.064 ± 0.014 0.5 mg / mL + 5% ethanol 0.064 ± 0.043 1 mg / mL + 5% ethanol 0.194 ± 0.051 **

CTL is untreated, * is p <0.05 and ** is p <0.01.

MTT activity when the fermented product of <Example 3> was treated division MTT activity (A ₅₇₀ ) CTL 0.261 ± 0.052 0 mg / mL + 5% ethanol 0.062 ± 0.041 0.125 mg / mL + 5% ethanol 0.069 ± 0.011 0.5 mg / mL + 5% ethanol 0.109 ± 0.012 1 mg / mL + 5% ethanol 0.362 ± 0.015 **

CTL is untreated group, * is p <0.05 and ** is p <0.01.

MTT activity when the fermented product of <Example 4> was treated division MTT activity (A ₅₇₀ ) CTL 0.261 ± 0.052 0 mg / mL + 5% ethanol 0.062 ± 0.041 0.125 mg / mL + 5% ethanol 0.113 ± 0.014 * 0.5 mg / mL + 5% ethanol 0.141 ± 0.032 ** 1 mg / mL + 5% ethanol 0.374 ± 0.012 **

CTL is untreated, * is p <0.05 and ** is p <0.01.

MTT activity when the fermented product of <Example 5> was treated division MTT activity (A ₅₇₀ ) CTL 0.261 ± 0.052 0 mg / mL + 5% ethanol 0.062 ± 0.041 0.125 mg / mL + 5% ethanol 0.156 ± 0.014 ** 0.5 mg / mL + 5% ethanol 0.172 ± 0.052 ** 1 mg / mL + 5% ethanol 0.453 ± 0.017 **

CTL is the untreated group, * is p <0.05 and ** is p <0.01. It can be seen that significant differences were observed at all concentrations of at least 1 mg / mL when the extracts or fermented products of the respective examples were treated. Here, the fermented product of <Example 5> had the highest liver cell protective activity.

Experimental Example 4 Fatty Liver Improvement Activity Experiment

In order to confirm the fatty liver improvement activity of the extract or fermented product of each Example, the lipid concentration and liver damage indicator enzyme activity of liver tissues of rats in which breeding was terminated in Experimental Example 1 were measured.

<4-1> Lipid concentration analysis of liver tissue

In order to investigate the change in lipid concentration of the liver according to the administration of the test substance, the rats of each experimental group in which the experiment was terminated in <Experimental Example 1> were extracted from the liver, and the washed liver was washed with physiological saline. (Folch J. et al., J. Biol. Chem., 226: 497-509, 1957), grind the liver finely, place it in a homogeneous tube, destroy the cell membrane with an ultrasonic cell membrane grinder, and remove chloroform and methanol ( Lipids were extracted with a mixture of chloroform: methanol = 2: 1). Then, the extract was centrifuged at 2000 rpm to obtain a supernatant, and the supernatant was used as a sample to measure the total cholesterol and triglyceride concentrations in the liver. The total cholesterol content was analyzed using a commercial kit (AM202-K, Asan Pharmaceutical Co., Ltd.). The absorbance was measured at 500 nm and the content was expressed in mg / dl according to the calibration curve. The triglyceride content was analyzed using a commercial kit (AM 157S-K, Asan Pharmaceutical Co., Ltd.). The absorbance was measured at 550 nm and the content was expressed in mg / dl according to the calibration curve.

The experimental results are shown in Tables 12 to 16 below.

Triglyceride and Cholesterol Concentrations in Liver Tissues of the Extract Administration Group of Example 1 division Triglyceride (mg / ㎗) Total Cholesterol Concentration (mg / dl) Normal group (Ⅰ) 39.78 ± 3.24 3.1 ± 0.9 Obesity control group (Ⅱ) 74.62 ± 1.17 8.2 ± 0.6 Experimental group (Ⅲ) 63.21 ± 2.35 ** 7.1 ± 0.4 **

* Is p <0.05 and ** is p <0.01.

Triglyceride and Cholesterol Concentrations in Liver Tissue of the Extract-administered Group of Example 2 division Triglyceride ㎎ / ㎗ Total cholesterol concentration mg / dl Normal group (Ⅰ) 38.57 ± 3.68 3.2 ± 0.8 Obesity control group (Ⅱ) 72.75 ± 3.43 7.9 ± 0.7 Experimental group (Ⅲ) 64.41 ± 3.23 ** 6.8 ± 0.7 **

* Is p <0.05 and ** is p <0.01.

Triglyceride and Cholesterol Concentrations of Liver Tissue in the Fermented Fermented Group of Example 3 division Triglyceride ㎎ / ㎗ Total cholesterol concentration mg / dl Normal group (Ⅰ) 37.57 ± 4.15 2.9 ± 0.9 Obesity control group (Ⅱ) 73.73 ± 2.76 8.3 ± 0.4 Experimental group (Ⅲ) 55.84 ± 1.24 ** 5.7 ± 0.9 **

* Is p <0.05 and ** is p <0.01.

Triglyceride and Cholesterol Concentrations of Liver Tissues from Fermented Fertilizers of Example 4 division Triglyceride ㎎ / ㎗ Total cholesterol concentration mg / dl Normal group (Ⅰ) 39.53 ± 2.57 3.0 ± 0.6 Obesity control group (Ⅱ) 71.72 ± 4.02 8.2 ± 0.7 Experimental group (Ⅲ) 52.31 ± 3.27 ** 5.5 ± 0.8 **

* Is p <0.05 and ** is p <0.01.

Neutral Fatty Acid and Cholesterol Concentrations of Liver Tissues of the Fermented Fermented Group of Example 5 division Triglyceride ㎎ / ㎗ Total cholesterol concentration mg / dl Normal group (Ⅰ) 35.05 ± 5.17 2.8 ± 1.1 Obesity control group (Ⅱ) 75.04 ± 1.27 8.4 ± 0.6 Experimental group (Ⅲ) 41.35 ± 1.29 ** 3.7 ± 0.9 **

* Is p <0.05 and ** is p <0.01. Both the extracts or fermented products of the above examples have the activity of lowering the concentration of triglycerides and cholesterol in liver tissue. The highest activity here is the fermented product of <Example 5>.

<3-2> Activity analysis of liver function indicator enzyme

GPT and GOT levels used as indicator enzymes of liver tissue damage were measured using an analysis kit (Asan Pharmaceutical, Korea) after separation of plasma after 97 days of experiment. > To <Table 21>.

Activity of liver function indicator enzyme of extract administration group of <Example 1> division GPT (IU / L) GOT (IU / L) Normal group (Ⅰ) 37.84 ± 4.23 67.45 ± 3.27 Obesity control group (Ⅱ) 63.74 ± 2.34 113.46 ± 1.55 Experimental group (Ⅲ) 57.56 ± 1.32 90.37 ± 3.21 **

* Is p <0.05 and ** is p <0.01.

Activity of liver function indicator enzyme of extract administration group of <Example 2> division GPT (IU / L) GOT (IU / L) Normal group (Ⅰ) 36.26 ± 2.54 65.64 ± 4.54 Obesity control group (Ⅱ) 65.21 ± 1.67 116.56 ± 5.21 Experimental group (Ⅲ) 57.05 ± 3.35 84.57 ± 5.32 **

* Is p <0.05 and ** is p <0.01.

Activity of liver function indicator enzyme of fermented product administration group of <Example 3> division GPT (IU / L) GOT (IU / L) Normal group (Ⅰ) 37.54 ± 3.15 68.94 ± 5.23 Obesity control group (Ⅱ) 62.42 ± 3.17 113.65 ± 4.87 Experimental group (Ⅲ) 52.21 ± 2.23 80.65 ± 1.21 **

* Is p <0.05 and ** is p <0.01.

Activity of liver function indicator enzyme of fermented product administration group of Example 4 division GPT (IU / L) GOT (IU / L) Normal group (Ⅰ) 39.27 ± 1.53 69.42 ± 2.32 Obesity control group (Ⅱ) 65.21 ± 1.34 116.45 ± 6.64 Experimental group (Ⅲ) 53.34 ± 2.37 81.75 ± 5.67 **

* Is p <0.05 and ** is p <0.01.

Neutral Fatty Acid and Cholesterol Concentrations of Liver Tissues of the Fermented Fermented Group of Example 5 division GPT (IU / L) GOT (IU / L) Normal group (Ⅰ) 37.94 ± 2.75 65.54 ± 3.56 Obesity control group (Ⅱ) 62.31 ± 3.52 119.76 ± 2.43 Experimental group (Ⅲ) 38.87 ± 4.25 69.43 ± 1.69 **

* Is p <0.05 and ** is p <0.01. It can be seen that the extract or fermented product of each of the above examples reduced GPT and GOT activity. In particular, the fermented product of <Example 5> has the highest effect.

Statistical processing

All the experimental results are the values represented by Mean ± SD. ANOVA analysis was performed on each experimental group, and significant differences were verified by Duncan test.

Claims (14)

A fatty liver improving food composition comprising a fermented product obtained by sterilizing and adding fermented rice bran by lactic acid bacteria and yeast to the dermis and fermented in a natural state as an active ingredient. The method of claim 1,
The lactic acid bacteria is a fatty liver improver food composition, characterized in that Lactobacillus genus lactic acid bacteria. The method of claim 1,
The yeast is genus Saccharomyces Fatty liver improver food composition, characterized in that it is yeast. The method of claim 1,
The amount of the fermented product of rice bran is added to the dermis fatty acid improver food composition, characterized in that 0.5 to 10 parts by weight based on 100 parts by weight of the dermis.  A food composition for improving obesity, comprising a fermented product obtained by sterilizing and adding fermented rice bran by lactic acid bacteria and yeast to the dermis and fermented in a natural state as an active ingredient. The method of claim 5,
The lactic acid bacteria is Lactobacillus genus lactic acid bacteria, characterized in that the obesity improver food composition. The method of claim 5,
The yeast is genus Saccharomyces Obesity improver food composition, characterized in that it is yeast. The method of claim 5,
The amount of the fermented rice bran is added to the dermis is an obesity improver food composition, characterized in that 0.5 to 10 parts by weight based on 100 parts by weight of the dermis. A food composition for diet comprising a fermented product obtained by sterilizing and adding fermented rice bran by lactic acid bacteria and yeast to the dermis and fermented in a natural state as an active ingredient. The method of claim 9,
The lactic acid bacteria is a diet food composition, characterized in that Lactobacillus genus lactic acid bacteria. The method of claim 9,
The yeast is genus Saccharomyces Dietary food composition, characterized in that the yeast. The method of claim 9,
The amount of the fermented rice bran is added to the dermis is a dietary food composition, characterized in that 0.5 to 10 parts by weight based on 100 parts by weight of the dermis. The method according to any one of claims 1 to 12,
The composition is a composition, characterized in that the beverage. Fatty liver improving pharmaceutical composition comprising a fermented product obtained by sterilizing and adding fermented rice bran by lactic acid bacteria and yeast to the dermis, and fermented product obtained by fermentation in natural state.