KR20160022961A - A composition for oral administration having anti-obesity effects containing Kimchi fermentation extracts - Google Patents

Abstract

The present invention relates to a composition for anti-obesity containing a kimchi fermentation extract. According to the present invention, a composition for oral administration containing, as an active ingredient, a kimchi fermentation extract obtained by fermenting and aging kimchi for 1-10 weeks and then extracting the same using water, ethanol, or a mixture thereof is shown to exhibit anti-obesity effects by inhibiting the expression of adipocyte differentiation factors, suppressing the expression of lipid metabolism-related factors and breaking intracellular neutral fats down into fatty acids and glycerol. Therefore, the kimchi fermentation extract can be variously used as a composition for oral administration or a food additive having anti-obesity effects. Moreover, it is expected to create added value of kimchi through the development of an anti-obesity agent material containing the kimchi fermentation extract.

Description

Technical Field [0001] The present invention relates to a composition for oral administration for anti-obesity containing kimchi fermentation broth extract,

The present invention relates to a composition for oral administration for anti-obesity containing kimchi extract. More specifically, the present invention relates to a composition for oral administration for anti-obesity containing Kimchi extract, which comprises fermenting the kimchi for 1 to 10 weeks and then extracting the mixture using water, ethanol, Which comprises the fermentation broth of Kimchi fermentation broth as an active ingredient. The present invention relates to a composition for oral administration for anti-obesity.

Obesity is reported to be not only a cosmetic problem but also a serious disease that can cause complications such as diabetes, hypertension, cardiovascular disease, cerebrovascular disease, arthritis, gallbladder disease, breast cancer and colorectal cancer. Many studies have been conducted to suppress the increase in obesity due to the decrease in physical activity due to development. This study was conducted to investigate the effect of dietary supplementation on body weight gain in rats fed high-calorie-diet. Korean Journal of Obstetrics and Gynecology Soc. Food Sci. Nutr., 33 (1), 91-100, 2004) used 1%, 5% and 10% of the feed intake of Chinese cabbage kimchi in white rats for 4 weeks using an oral zonde (Kim et al., 2003), which has been reported to be effective in reducing the weight loss of 5% A "kimchi biogenic formula" was prepared and given to white rats, 17% subcutaneous fat reduction and 21% plasma triglyceride lowering effect were reported. Baek, YH, Kwak, JR, Kim, SJ, Han, SS, Song YO, Effects of kimchi supplementation and / or exercise training on body composition and plasma lipids in obese middle school girls J. Korean Soc. Nutr., 30 (5), 906-912, 2001) reported that 8% body fat reduction was obtained when kimchi supplements were consumed in obese middle school students over BMI 25 for 6 weeks.

Therefore, the present inventors have continued to investigate what constituents of kimchi can provide an obesity-suppressing effect. As a result, the present inventors have found that kimchi, preferably fermented kimchi, more preferably fermented cabbage kimchi, The present inventors have completed the present invention by discovering that an anti-obesity effect can be expected from an ethanol-extracted cabbage kimchi extract after a fermentation process.

Therefore, a technical problem to be solved by the present invention is to provide a composition for oral administration for anti-obesity comprising a kimchi extract.

In order to solve the above technical problems, the present invention provides a kimchi fermentation broth extract obtained by fermenting and aging kimchi for 1 to 10 weeks and then extracting it with water, ethanol or a mixture thereof, For oral administration.

The composition for oral administration comprising the extract of Kimchi fermentation broth according to the present invention suppresses the expression of adipocyte differentiation factor and inhibits the expression of lipid metabolism-related factors as well as the anti-obesity effect by decomposing intracellular triglyceride into fatty acid and glycerol . Therefore, it can be used variously as a composition for oral administration or a food additive having an anti-obesity effect including the kimchi fermentation broth extract. In addition, it is expected that the added value of kimchi will be possible through the development of anti - obesity material including the kimchi fermentation broth extract.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments of the invention and, together with the description of the invention, It should not be construed as limited.
FIG. 1 shows the degree of fat accumulation of the Kimchi fermentation broth extract, wherein FK is a 70% ethanol extract of an early fermented kimchi, OFK is a 70% ethanol extract of the kimchi fermentation broth of the present invention, and LFK is 70% KW is the water extract of the kimchi fermentation broth of the present invention, and CK is the 70% ethanol extract of the total kimchi fermentation broth.
FIG. 2 is a graph showing the content of triglycerides by Kimchi fermentation broth extract. KE is a 70% ethanol extract of Kimchi fermented product of the present invention.
FIG. 3 is a graph showing the expression of an adipocyte differentiation factor by kimchi fermentation broth extract, wherein KE is a 70% ethanol extract of kimchi fermented product of the present invention.
FIG. 4 is a graph showing the expression of fat-metabolism-related factors expressed by the kimchi fermentation broth extract. KE is a 70% ethanol extract of the fermented kimchi of the present invention.
FIG. 5 is a graph showing the expression level of an enzyme factor related to fat cell degradation by Kimchi fermentation broth extract. KE is a 70% ethanol extract of kimchi fermented product of the present invention.

The composition for oral administration for anti-obesity of the present invention is characterized in that the kimchi is fermented and aged for 1 to 10 weeks, and then extracted with water, ethanol or a mixture thereof, as an active ingredient .

Preferably, the kimchi fermentation broth extract may be prepared by a method comprising the steps of:

(S1) submerging the kimchi and fermenting for 1 to 10 weeks, followed by lyophilization;

(S2) extracting the lyophilized kimchi with water, ethanol or a mixture thereof for 20 to 30 hours; And

(S3) The step of filtering the extracted Kimchi extract using a filter paper followed by centrifuging to remove the precipitate, concentrating and freeze-drying the precipitate.

The kimchi used in the present invention includes any of kimchi such as Chinese cabbage kimchi, mugi kimchi, mustard kimchi, radish kimchi, and radish kimchi, but cabbage kimchi is particularly preferable.

The fermentation of the kimchi in step (S1) in the step of producing the Kimchi extract of the present invention is characterized by being performed for 1 week to 10 weeks, preferably 2 weeks to 8 weeks, more preferably 4 weeks, Is carried out at -50 to -80 占 폚, preferably? 70 占 폚.

The storage stability of the kimchi fermentation broth can be improved by performing the lyophilization step, and it is advantageous that the kimchi fermentation broth is more suitable and easy to be mass-extracted in the subsequent step.

As the extraction solvent for the Kimchi extraction in (S2), water, ethanol or a mixture thereof may be used, and ethanol is particularly preferable. The amount of the extraction solvent for the kimchi is 2 to 4 L, preferably 3 L per 10 g of the kimchi, and the extraction time is 20 to 30 hours, preferably 24 hours.

When the fermentation period is less than one week, the anti-obesity effect of the kimchi extract is lowered. If the fermentation period is longer than 10 weeks, the anti-obesity effect does not occur in proportion to the fermentation time.

The Kimchi extract thus obtained has a concentration of 50 to 1,000 占 퐂 / ml, preferably 100 to 500 占 퐂 / ml, which is most suitable for anti-obesity effect.

The kimchi used in the present invention includes any kimchi prepared according to a conventional kimchi production method or a commercially available kimchi, and there is no particular limitation in the production method thereof. For example, it can be prepared by a method including the following steps.

(1) pickling the Chinese cabbage in pickled water for 10 to 12 hours, then rinsing and draining;

(2) Washing the radish, grinding it in a mixer, squeezing it with berubo, mixing it with clear juice, gad and safflower;

(3) putting broth in red pepper powder and boiling for 10 to 20 minutes, then adding chopped garlic and chopped ginger, and adding kimchi, safflower and mustard prepared in (2)

(4) placing the kimchi in a state where the pickled Chinese cabbage is turned upside down in step (1); And

(5) Put kimchi into a bucket so that the cutting surface is on top, then cover the outer leaves of the pickled cabbage and spread the salt.

The composition for anti-obesity of the present invention may contain the fermentation broth extract of Kimchi in an amount of 0.1 to 100% by weight, preferably 1 to 70% by weight, based on the total weight of the composition.

The composition for oral administration comprising the extract of Kimchi fermentation broth of the present invention inhibits the expression of adipocyte differentiation factor and inhibits the expression of fat metabolism-related factors as well as an anti-obesity effect by decomposing intracellular triglyceride into fatty acid and glycerol . Therefore, it can be usefully used as a composition for oral administration having an anti-obesity effect including the kimchi fermentation broth extract.

The pharmaceutical composition of the present invention containing the Kimchi fermentation broth extract of the present invention shows no toxicity such as acute toxicity in the test using the rats and shows no adverse effect on liver function.

The pharmaceutical composition of the present invention can be used to prepare pharmaceutical formulations according to a conventional method. In the production of formulations, it is preferable to mix or dilute the fermented extract of Kimchi, which is an active ingredient, together with the carrier, or enclose the carrier in the form of a container. When the carrier is used as a diluent, it may be a solid, semi-solid or liquid substance which acts as a vehicle, excipient or medicament for the active ingredient.

Thus, the formulations may be in the form of tablets, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, soft or hard gelatin capsules, sterile powders and the like. Examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, Propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The formulations may additionally include fillers, anti-coagulants, lubricants, wetting agents, perfumes, emulsifiers, preservatives, and the like. The pharmaceutical compositions according to the present invention can be formulated using methods well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal.

The pharmaceutical composition according to the present invention can be administered through various routes including oral, transdermal, subcutaneous, intravenous or muscular. In the case of humans, the usual daily dose may be in the range of 1 to 1,000 mg / kg body weight, preferably 10 to 100 mg / kg body weight, and may be administered once or several times in divided doses. However, the actual dosage should be determined in light of various relevant factors such as the route of administration, the age, sex and weight of the patient, the health condition, and the severity of the disease.

Meanwhile, the composition for anti-obesity comprising the Kimchi fermentation broth extract of the present invention can be usefully used as a food additive.

The food additive (or food additive) refers to a substance that is added to food, mixed, infiltrated or otherwise used for the manufacture, processing, or preservation of food. With the development of culture and the affordability of the economy, the quantity of food increased, the quality was improved, and the demand for convenience increased. As a result, mass-produced processed foods, instant foods, etc. are increasingly on the table, and new kinds of foods are pouring out every day, and this phenomenon is accompanied by the use of food additives.

There are two types of food additives, natural and synthetic, and they are usually classified according to their functions and uses. Currently, about 370 kinds of chemical synthetic products and 50 kinds of natural additives are approved as food additive in Korea. Depending on the application, preservatives, disinfectants, antioxidants, coloring agents, coloring agents, bleaching agents, seasonings, sweeteners, flavoring agents, , Emulsifiers, thickeners (thickeners) and stabilizers, encapsulating agents, gum base agents, defoamers, solvents, release agents, insect repellents, quality improvers and other food additives. On the other hand, functional food refers to food that has been developed so that the third function (biological function), which is a nutritional function as a primary function of food, a sensory function as a secondary function, Functional foods directly contribute to the improvement of quality of life by preventing chronic diseases by biological control function, alleviation of individual and national medical expenses, and the formation of an affluent welfare state through the implementation of health society. Biocontrol functions that the food may exhibit include anticancer, blood pressure lowering, cholesterol lowering, blood thrombosis inhibition, prevention of diabetes, and inhibition of aging, and materials having specific functions may be used as food additives in order to impart such functionality to foods.

As the composition for oral administration containing the Kimchi fermentation broth extract of the present invention, the food additive may be an organic acid such as citric acid, fumaric acid, adipic acid, lactic acid, and malic acid; Phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate, and polyphosphate (polymerized phosphate); May contain one or more selected from natural antioxidants such as polyphenol, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid and phytic acid .

It may further comprise one or more selected from lactose casein, dextrin, glucose, sugar, sorbitol.

In addition, the present invention provides, as a preferred embodiment, a food comprising the above-mentioned composition for oral administration.

The food of the present invention may contain other additives such as preservatives, bactericides, antioxidants, spices, seasonings, sweeteners, flavoring agents, swelling agents, enhancers, improvers, emulsifiers, various nutrients, flavors such as synthetic flavors and natural flavors, , Coloring agents, thickening agents (cheese, chocolate etc.), pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, defoamers, solvents, release agents, preservatives, , And other food preparation additives such as carbonating agents.

The food additives and other food additives can be added to the food by immersion, spraying or mixing with the food.

The food according to the above embodiment may be one or more selected from beverage, chips, noodles, processed foods, cereals, tablets, soft capsules and granules.

More specifically, the food is a dried product of fruits, vegetables, fruits or vegetables; Or cut products, fruit juices, vegetable juices or mixed juices thereof; One or more of chipping, noodles, livestock processed food, marine processed food, milk processed food, fermented milk food, cereal food, cereal food, fermented microorganism food, confectionery bakery, condiments, meat processing industry, acid drink, licorice, .

For example, when preparing gum, 0.5% by weight of kimchi fermentation broth extract, 25% by weight of gum base and 72% by weight of sorbitol are mixed in a preheated mixer at 60 to 70 DEG C, and the mixture is kept at 55 DEG C while peppermint flavor 2.5% by weight, then mixed for 30 minutes, extracted, rolled, aged for 24-48 hours and then cut and packed to produce gum.

It can also be added to kneading in confectionery baking process, added to sauces and sauces, and added as powdered or liquid form to fermented milk or yogurt. It is also possible to use the herbal medicine extract powder for the purpose of taste preservation, antioxidation and discoloration prevention by directly putting it into kimchi, salted fish and vegetable processing. The food additives according to the present invention in the form of powder or granules can be mixed with garlic, ginger, herb, licorice and the like to make spices and used in fish processing It can also be used for immersion, spraying and addition methods such as fish paste, fish, seasoned cane, and fermented fish, and can be added by immersion, spraying and mixing in kimchi, radish, pickled mackerel and pickled vegetables.

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

Example 1 Preparation of Ethanol Extract of Fermented Kimchi

It was prepared according to the following method as a laboratory recipes containing no salted fish.

As a main ingredient, 10 kg of Chinese cabbage (4 drops) and 500 g of rice (1/2) were prepared and mixed with 7 cups of salted salty salt and 30 cups of water. As fresh ingredients, prepare freshly 4-stem and 10-stem, and prepare 2 cups of red pepper powder, 1/2 cup of broth, 5 tablespoons of chopped garlic, 3 tablespoons of chopped ginger and 7 tablespoons of salt.

(1) Chinese cabbage pickle

Pick a medium-sized Chinese cabbage about 2.5 ~ 3 ㎏, remove the leaves and cut into a small amount of pickled salt water. If the cabbage is large, it is cut into four halves. The basic pickles were prepared by mixing 0.25 kg of sun salmon per kg of Chinese cabbage and 1.2 kg of water.

Turn it upside down in the middle and leave at room temperature for 10 ~ 12 hours. Pickled cabbage is rinsed 3 to 4 times in running water, then it is turned over for about 1 hour and drained.

(2) No preparation

I rinse the bark cleanly and bake it in the mixer, and I just get clean squeezed juice. Wash god and safflower and cut into 3㎝ length.

(3) Preparing Kimchi

Put red pepper in the red pepper, put it for about 20 minutes, then chopped garlic and chopped ginger. Add safflower and mustard and mix with salt.

(4) Insert Kimchi

Turn the pickled cabbage upside down and insert the kimchi cow from the back. Only put the cow in the stem and wrap it in the outer leaves.

(5) in a container

The cabbage cuts into the pail so that the cutting surface is on the top, then covers the outer leaves of the pickled cabbage and sprinkles with salt.

The prepared Chinese cabbage kimchi was fermented at 10 캜 for 4 weeks, stored at -70 캜 for freeze drying, and stored. After lyophilization, 20 g of each was put in a flask, and 200 ml of 70% ethanol was added thereto. The mixture was mixed well at room temperature for 24 hours, and then the filter was repeated three times. The solution obtained by repeating 3 times was concentrated under reduced pressure, and then lyophilized for 72 hours and dissolved in DMSO at a concentration of 50 mg / ml.

≪ Comparative Example 1 > Preparation of fermented Kimchi water extract

A kimchi fermentation broth extract was prepared in the same manner as in Example 1, except that the kimchi fermentation broth was extracted with water instead of ethanol.

≪ Comparative Example 2 >

For the preparation of the fermented product, the kimchi containing no fermented fish was prepared in the same manner as in Example 1, and the fermented kimchi was preserved by freeze-drying at 70 ° C on the same day without fermentation. Ethanol extraction was carried out in the same manner as in Example 1.

≪ Comparative Example 3 > Preparation of ethanol extract of mukjji

The matured (fermented for 12 months) was lyophilized at < RTI ID = 0.0 > 70 C < / RTI > and stored, followed by ethanol extraction in the same manner as in Example 1 above.

<Comparative Example 4> Preparation of ethanol extract of bamboo kimchi

The commercially available bamboo kimchi was stored at &lt; RTI ID = 0.0 &gt; 70 C &lt; / RTI &gt; by lyophilization and ethanol extraction was carried out in the same manner as in Example 1 above.

&Lt; Test Example 1 > Effect of Kimchi fermentation broth extract on differentiation of adipocytes

The 3T3-L1 preadipocyte cell line was cultured and differentiated in order to examine the effect of kimchi fermentation step and extraction solvent on differentiation of adipocytes prepared in Example 1 above.

(1) induction of culture and differentiation of 3T3-L1 whole fat cell line

The 3T3-L1 preadipocyte cell line was cultured in DMEM (Dulbecco's Modified Eagle Medium, Gibco) containing 10% CS (Calf Serum), 100 units / ml of penicillin and 100 ng / ml of streptomycin, And cultured in an incubator maintained with CO ₂ . At this time, the cell surface was washed with PBS (phosphate buffered saline) solution every time the medium was exchanged, and the culture medium was exchanged with a DMEM culture medium containing fresh 10% CS medium. When cells were adhered to the bottom of the flask by 80% as a single layer, the cell surface was washed with PBS solution, and 0.25% trypsin-EDTA (Gibco) solution was added and centrifuged. These cells were divided into 2 × 10 ⁵ cells in a 6-well plate and cultured in DMEM containing 10% FBS (Fetal Bovine Serum), 100 units / ㎖ penicillin and 100 ng / ㎖ of streptomycin (Dulbeeco's Modified Eagle Medium, Gibco) were used. Two days after 100% confluent stage, DMEM containing 10% FBS was replaced with culture medium containing 0.05 μM Dexamethasone, 0.25 mM IBMX, and 2 μg / mL insulin to induce differentiation for 3 days. After 3 days, the cells were treated with DMEM supplemented with 10% FBS containing insulin alone for 3 days, and DMEM supplemented with 10% FBS was used for the final 3 days.

(2) Statistical processing

Statistical significance was tested using the GraphPad Prism 5 program and the difference between groups was statistically analyzed using Bonferroni post hoc test of one-way ANOVA. The significance was tested at p <0.05 level.

(3) Fat accumulation measurement by oil red o staining

The differentiation of adipocytes in the lipid differentiation process among the ethanol extract of the fermentation broth of Example 1, the water extract of the kimchi fermentation broth of Comparative Example 1, the initial kimchi of Comparative Example 2, the mung bean extract of Comparative Example 3 and the bamboo kimchi extract of Comparative Example 4 And the amount of fat accumulation was measured using Oil red O staining method to select effective fractions and samples. In order to select the effective fractions and samples for the inhibition of adipocyte differentiation among the ethanol extracts and the water extracts of mungbean kimchi, the extracts of 70% ethanol and water were treated at a concentration of 500 ㎍ / ㎖ to induce differentiation And the degree of fat accumulation was measured using Oil red O staining method. The cells were treated with ethanol extract and water extract at a concentration of 500 ㎍ / ㎖ for 2 days after the cells were 100% warmed on the plate, when the differentiation inducing reagent (IBMX, DEX, INS) supplemented culture medium, insulin supplemented culture medium and FBS culture medium were changed. After 9 days of differentiation, the culture medium was removed, washed with PBS, and fixed with 10% formaldehyde for 10 minutes. 10% formaldehyde was removed and the cells were fixed for more than 1 hour by adding saturated formaldehyde again. Then, 60% isopropanol was added to the solution, and the solution was immediately washed with distilled water, stained with oil red O staining solution for 30 minutes, and washed with distilled water. The intracellular fat accumulation of the dysplasia was observed using a microscope. The photographs showed that the ethanol extract of Kimchi fermentation broth of Example 1, the water extract of Kimchi fermentation broth of Comparative Example 1, the initial Kimchi of Comparative Example 2, and Comparative Example 3 And the effect of kimchi extract of Comparative Example 4 on the fat accumulation during fat differentiation were observed. To confirm the accumulation of fat, isopropanol was added to the dried oil, and the oil red O dye was eluted. The absorbance was measured at 500 nm using an absorbance meter.

The results are shown in Fig. The ethanol extract of the Kimchi fermentation broth of Example 1 was more effectively reduced in fat accumulation than the control Kimchi, muconic acid and bamboo kimchi of Comparative Examples 2, 3 and 4 and the Kimchi fermentation broth of Comparative Example 1. Specifically, the initial kimchi ethanol extract of Comparative Example 2 when the control group was set at 100%; 75.18%, the Kimchi fermentation liquid ethanol extract of the present invention; 43.99% ethanol extract of Comparative Example 3; 113.4%, the bamboo kimchi ethanol extract of Comparative Example 4; 86.47%, the Kimchi fermentation water extract of Comparative Example 1; 73.91%. 1, GM; Growth medium, DM; Differentiation medium, FK; Early Kimchi Ethanol Extract, OFK; Kimchi fermentation liquid ethanol extract (KE), LFK; How old are you, CK; Bamboo kimchi ethanol extract, KW; Kimchi fermented liquid water extract.

(4) Triglyceride assay

Neutral fat assay kit was used to quantitatively measure TG in differentiated cells as a fluorescent stain that can easily measure the amount of triglyceride in the cytoplasm. On the 9th day, the media was removed, washed twice with D-PBS, treated with reagent for 1 hour, and then absorbed at 570 nm.

The results are shown in Fig. Most lipids are composed of proteins such as TG and perilipin A. In order to more precisely determine the effect of the selected sample and its ethanol extract on the fat accumulation by the oil-red O staining method, the TG content, which is the main component of the lipids contained in the cells, was measured Respectively.

In the differentiation of 3T3-L1 preadipocytes into adipocytes, the selected kimchi fermentation broth was treated with 50, 250 and 500 ㎍ / ㎖ of ethanol extracts to induce differentiation and triglyceride content was measured. The ethanol extract of kimchi fermentation broth of the present invention showed a significant decrease of triglyceride concentration in the concentration of 50 ㎍ / ㎖, 86.02%, 250 ㎍ / ㎖ 41.55%, and 500 ㎍ / ㎖ 16.39%, respectively. As a result of measuring the triglyceride content according to the differentiation of adipocytes, at the concentration of 500 μg / ml, more than 80% of the triglyceride was decreased compared to the control group.

(5) Measurement of expression level of adipocyte differentiation factor by ethanol extract

The recovered cells were washed with PBS, suspended in a solution containing RIPA buffer and protease inhibitor cocktail (Roche), eluted for 15 minutes, disrupted with an ultrasonic crusher, and centrifuged at 13,000 rpm for 20 minutes . The supernatant was quantified by BCA method and the same amount of protein (20 μg) was electrophoresed on SDS-polyacrylamide gel. After electrophoresis, proteins in the gel were transferred to a PVDF membrane, and then blocked with a nonspecific protein blocking solution (0.15 M NaCl, 1 M Tris-HCl (pH 7.5), 0.1% Triton X-100, 5% Skim milk) For 3 hours at RT. Membranes were washed and primary antibodies were treated and then reacted with secondary antibodies suitable for the primary antibodies treated. Immunoreactive protein was detected with a chemiluminescence system (G: BOX Range Fluorescence Manual Image Capture; SuperSignal West Femto Maximity Sensitivity Substrate, Pierce, USA). The primary antibodies of PPARγ, C / EBPα, AMPKα, SREBP-1c, FAS, CD36, GLUT4 and AP2α were purchased from Cell Signaling Technology (USA) mouse IgG-HRP, anti-rabbit IgG-HRP and anti-goat IgG-HRP were purchased from Santa Cruz (Santa Cruz, CA, USA). In the process of differentiation into adipocytes from the cell follicle, biochemical changes such as the expression of adipocyte-specific protein markers, as well as the morphological changes of lipid peroxidation by fat accumulation, appear, and the transcription factors associated with adipocyte differentiation Expression appears. The most important active factors are PPARγ and C / EBPα. In order to determine the molecular mechanism by which the decrease of lipid accumulation by 3T3-L1 preadipocyte differentiated into the adipocytes was induced by the ethanol extract of Kimchi fermentation broth, 70% ethanol extracts of 50, 250 and 500 ㎍ / ㎖ , And the effect on PPARγ, C / EBPα, which is involved in differentiation after induction of differentiation, was observed. The results are shown in FIG.

As a result, the concentration of ethanol extract of Kimchi fermentation broth tended to decrease in comparison with that of the control group, so that the expression of transcription factors could be suppressed and the production of intracellular TG could be decreased.

(6) Measurement of expression level of fat metabolism-related factor by ethanol extract

Fat cells play an important role in maintaining energy homeostasis and lipid metabolism. 3T3-L1, a lipogenic precursor cell, is differentiated into adipocytes by various hormones and various transcription factors and accumulates intracellular fat. Differentiated cells have biochemical features that induce expression of adipocyte-specific genes such as FAS, SREBP and aP2, as well as morphological features such as accumulation of triglyceride in white adipocytes. To differentiate 3T3-L1 preadipocytes into adipocytes, we investigated the effect of ethanol extract of Kimchi fermentation broth on the lipid metabolism-related factors by inducing differentiation by treatment with selected concentrations of 50, 250 and 500 ㎍ / ㎖ The expression levels of AMPKa, SREBP-1, FAS, CD36, Glut4 and aP2, which are involved in lipid metabolism, were measured and the results are shown in FIG. The concentration of AMPKα, SREBP-1, FAS, CD36, Glut4, and aP2 in the ethanol extract of Kimchi fermentation broth tended to decrease in a concentration-dependent manner. In particular, SREBP-1, FAS and aP2 showed strong inhibitory effects on lipid metabolism-related factors. This suggests that the inhibition of the expression of the differentiation factor as well as the expression of the fat metabolism - related factor may reduce the production of intracellular TG.

<Test Example 2> Effect of ethanol extract of kimchi fermentation broth on fat cell metabolism

(1) Determination of the expression levels of PKA, HSL, ATGL and LPL in the differentiated 3T3-L1 adipocytes following treatment with the ethanol extract of kimchi fermentation broth

After 3T3-L1 adipocyte differentiation, western blot assay was performed to determine the effect of ethanol extract of kimchi fermentation broth on lipid peroxidation protein expression level. Mature adipocytes were treated with ethanol extract of kimchi fermentation broth at various concentrations and the expression of lipolytic enzymes was measured 48 hours later in mature adipocytes treated with control and extracts.

Lipopolysaccharide (LPL), HSL and ATGL are known to be involved in lipid metabolism. LPL is an extracellular enzyme mainly distributed in adipose tissue, cardiac tissue, and capillary around the skeletal muscle, which decomposes triglyceride in chyromicrone and separates it into monoglyceride and two chain fatty acids And it facilitates the entry into the cell, thereby contributing to the reproduction of triglyceride in adipocytes. HSL, on the other hand, is known to be an enzyme that is activated by phosphorylation by PKA activated by the action of adrenergic hormone to decompose triglycerides present in cells and produce fatty acids and glycerol. The resulting glycerol is released out of the cell. ATGL is known to be involved in the production of diglycerides, the first step in the degradation of triglycerides. Therefore, it can be said that lipid accumulation and degradation are controlled to a certain level by these lipolytic enzymes. In order to investigate the mechanism by which fat accumulation was reduced and triglyceride was decomposed and released into free glycerol in the differentiated 3T3-L1 adipocytes, the extracts of Kimchi fermentation broth were treated with 250, 500, 1,000 ㎍ / Next, the expression level of lipase was observed and the results are shown in Fig. The expression level of PKA and LPL was not significantly different from that of the control group. As a result of observing the expression level of HSL, the concentration-dependent decrease was observed, and the expression level of phospho-HSL was observed to increase in a concentration-dependent manner. In particular, ATGL expression tended to increase in a concentration-dependent manner. This suggests that the mechanism of phosphorylation of HSL and the expression of ATGL may result in a decrease in fat accumulation. The more influential tendency is seen. The expression of HSL and ATGL genes in lipolytic enzymes except LPL seems to be decomposed into fatty acids and glycerol in the intracellular triglyceride.

The composition for oral administration comprising the extract of Kimchi fermentation broth according to the present invention suppresses the expression of adipocyte differentiation factor and inhibits the expression of lipid metabolism-related factors as well as the anti-obesity effect by decomposing intracellular triglyceride into fatty acid and glycerol . Therefore, it can be used variously as a composition for oral administration or a food additive having an anti-obesity effect including the kimchi fermentation broth extract. In addition, it is expected that the added value of kimchi will be possible through the development of anti - obesity material including the kimchi fermentation broth extract.

Claims (12)

Characterized in that the kimchi is fermented and aged for 1 to 10 weeks, followed by lyophilization, and then extracted with water, ethanol or a mixture thereof, as an active ingredient. Composition. The method according to claim 1,
Wherein the kimchi is a cabbage kimchi. The method according to claim 1,
Wherein the extract of the fermentation broth of the kimchi is contained in an amount of 0.1 to 100% by weight of the total weight of the composition. The method according to claim 1,
Wherein the kimchi fermentation broth extract is prepared by a method comprising the steps of:
(S1) submerging the kimchi and fermenting for 1 to 10 weeks, followed by lyophilization;
(S2) extracting the lyophilized kimchi with water, ethanol or a mixture thereof for 20 to 30 hours; And
(S3) The step of filtering the extracted Kimchi extract using a filter paper followed by centrifuging to remove the precipitate, concentrating and freeze-drying the precipitate. 5. The method of claim 4,
Wherein the fermentation of the kimchi is performed for 2 to 8 weeks in the step (S1). 6. The method of claim 5,
Wherein the fermentation period is 4 weeks. 5. The method of claim 4,
Wherein the kimchi is freeze-dried at -50 to -80 占 폚 in the step (S1). 5. The method of claim 4,
Wherein the extract is prepared by using 2 to 4 L of water, ethanol or a mixture thereof per 10 g of the kimchi in the step (S2). 5. The method of claim 4,
Wherein the ethanol is used in the step (S2). The method according to claim 1,
Wherein the kimchi fermentation broth extract has a concentration of 50 to 1,000 占 퐂 / ml. The method according to claim 1,
Wherein the kimchi is prepared according to the following process:
(1) pickling the Chinese cabbage in pickled water for 10 to 12 hours, then rinsing and draining;
(2) Washing the radish, grinding it in a mixer, squeezing it with berubo, mixing it with clear juice, gad and safflower;
(3) putting broth in red pepper powder, boiling for 10 to 20 minutes, adding chopped garlic and chopped ginger, mixing the soup, safflower and mustard prepared in (2) above and salt to make Kimchi;
(4) placing the kimchi in a state where the pickled Chinese cabbage is turned upside down in step (1); And
(5) Put kimchi into a bucket so that the cutting surface is on top, then cover the outer leaves of the pickled cabbage and spread the salt. 12. A food additive comprising the composition for oral administration according to any one of claims 1 to 11.

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