KR101501233B1 - Pharmaceutical composition or health food containing black tea ethanol solution extracts which is effective for reducing body weight and body fats or preventing or treating lipid related metabolic disease - Google Patents

Abstract

The present invention relates to a composition containing a black tea extract. More particularly, the herbal medicine extract extracted with the aqueous ethanolic solution of the present invention is more effective than the herbal medicine extract obtained by conventional hot water extraction and ethanol heating extraction methods, The present invention relates to a method for preventing and treating obesity, metabolic syndrome, diabetes and hyperlipidemia, and a method for preventing obesity, metabolic syndrome, diabetes, and hyperlipidemia, by increasing the content of gallic acid, Can be usefully used as an active ingredient of a pharmaceutical composition useful for treatment and a health food.

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition and a health food for preventing or ameliorating body weight and fat loss or lipid-related metabolic diseases containing a black tea ethanol aqueous solution cold extract, or preventing or treating lipid related metabolic disease}

TECHNICAL FIELD The present invention relates to a composition containing a black tea extract aqueous solution of a black tea ethanol aqueous solution, and more particularly to a composition for anti-obesity containing a black tea extract aqueous solution of black tea ethanol or a composition for preventing, improving or treating a lipid-related metabolic disease.

Obesity is a biological phenomenon caused by the interaction of complex factors such as genetic, metabolic, environmental, and behavioral factors, and is generally recognized as excess weight. Medically speaking, the body mass index (BMI) Or more than 30% of body weight), a BMI of 27 or more, and other circulatory diseases such as diabetes, hypertension, and hyperlipidemia. In particular, obesity is an important cause of various adult diseases associated with insulin resistance, diabetes, hypertension, cancer, gallbladder disease, hyperlipidemia, arteriosclerosis and the like. In addition, in obese patients and animals, Which has recently become known as a cause of social concern. The cause of obesity known to date is known to be more than 70% of the genetic predisposition, and other environmental factors such as high fat diet intake and lack of exercise are known. Recently, however, attention has been focused on the amount of energy consumed and the amount of energy consumed Is gathering. In other words, it is difficult to see genetic causes only because the incidence has rapidly increased even though the genetic predisposition did not change much over the last decades. Therefore, genetic and environmental complex factors destroying energy balance are important factors in weight determination It is recognized as an important factor. Therefore, accurate identification of the cause of obesity is urgently required.

Currently known herbal remedies include Xenical (Roche Pharmaceuticals, Switzerland), Reductil (Evothe, USA) and Exolise (Atopama, France), which are widely used as appetite suppressants, And most obesity drugs are appetite suppressants that suppress appetite by controlling the neurotransmitters associated with the hypothalamus. However, the present therapeutic agents have side effects such as heart diseases, respiratory diseases, nervous system diseases and the like, and the persistence of their effects is also low, so that it is necessary to develop more improved therapeutic agents for obesity. Also, currently developed products have satisfactory therapeutic effects There is no cure, and development of new obesity treatment is required.

The lipid-related metabolic disease refers to a blood lipid-related disease among disorders caused by metabolic disorders in vivo, and specifically includes fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, atherosclerosis and lipid-related metabolic syndrome. The metabolic syndrome refers to a disease in which various metabolic diseases such as diabetes and obesity occur simultaneously in one person. The above-mentioned lipid-related metabolic diseases are also required to develop effective therapeutic agents as well as obesity.

On the other hand, black tea is fermented tea after fermentation by microorganism in tea plant belonging to Theaceae ( Camelia sinensis L.), gallic acid, caffeine, It contains catechins. There are many kinds of black tea in China depending on the place of production. Bochaja, Chiljae, and other cars are produced in Yunnan Province. The strong cars and gold coins are produced in Sichuan province, the blast furnace cars, black tanks, It's a car. Boi cha, a typical black tea, is a tea made by using a tea made by drying in the sun using Yunnan Taisha-jong tea leaf which the minority peoples of China began to drink for a long time. Minority peoples were mainly known as the mainland to drink and in 1726 it was designated as a tolerance. Depending on the shape, there are long-distance cars made of straw tea, steamed tea and lump, and various types of long tea are distributed in various places such as a bottle, a tank, a long car, a braking car, and others. The tea, made by the natural drying method, was almost lost during the Cultural Revolution. Currently, most of the circulating products are microorganisms produced by using the manufacturing process called 伤 퇴 (堆 堆 퇴). There is an increasing tendency to use it as the fact that it has the effect of decomposing fat and cholesterol (Food Science Dictionary, Doosan Encyclopedia Dipipedia).

The common production process of Boi tea is the process of picking tea leaves, firing (drying in a cool place to reduce the moisture content of tea leaves), salting (killing blue tea leaves) In the process, the tea leaves are turned green in a large pot, and the tea leaves are dark green.), Brewing tea, and drying tea leaves to promote fermentation. If you use tea leaves in a bamboo seat and spread it in the sun, the process of drying in the sun / 청 악 악 (((((渥 4 The color of the matured tea is reddish, the tea does not go through this process), the process of pressing (lastly molding and compressing to form a shape such as a bottle or a car) Hit.

On the other hand, Korean Patent No. 10-0443116 discloses a fermented tea obtained by fermenting tea leaves collected from a broadleaf tea plant native to Yunnan, China for 4 years or more in a dry or wet fermentation process, in an amount of 20% aqueous solution of 50% ethanol (w / w) And the composition containing the extract is effective for the prevention and treatment of obesity and hyperlipidemia by exhibiting pharmacological activity of antiobesity and blood cholesterol lowering effect.

In addition, Yamagami T. et al. Reported that the Chinese black tea hot water extract reduced the total cholesterol and LDL cholesterol (Ann Nutr Metab, 53 (2008) 33-42) and the water soluble extract of Chinese black tea Hidenori Urata et al. Reported that the aqueous extract of Chinese black tea improved the obesity index in humans and decreased the abdominal fat (Nutrition Research, 28 (2008) 450 ~ 456) Research, 31 (2011) 421-428). However, the effect of the above-mentioned papers corresponds to the extracts extracted by hot water extraction or ethanol heat extraction. On the other hand, gallic acid, catechins, etc., which are known to be effective components of black tea, show a decrease in content under high temperature solution conditions. In particular, gallic acid (British journal of Nutrition 98 (2007), 98, 727-735, Chemico-Biological Interactions, 174 (2008), 109-117), which has efficacy for weight loss and fat reduction, (Geochimica et Cosmochimica Acta, Vol. 52, pp. 341-344). The content of catechin, which is known as an effective ingredient of black tea, also shows little change at 24 ° C to 37 ° C, but it is decreased by 20 to 25% at 100 ° C for 3 hours (Stability of tea theaflavins and catechins, Food Chemistry , Vol. 83 pp. 189-195).

Accordingly, the present inventors have found that the black tea extract extracted with the ethanolic aqueous solution of the present invention has an increased content of gallic acid, which is an active component of the black tea and an indicator component thereof, as compared with the black tea extract extracted by the conventional hot water extraction and ethanol heating extraction method , And the present inventors completed the present invention by confirming that the high fat diet method has excellent effects on weight and body fat reduction, fatty liver improvement, blood glucose control and cholesterol reduction in an obesity mouse model.

It is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of obesity or lipid-related metabolic diseases, which comprises an extract of black tea ethanolic aqueous solution as an active ingredient.

It is still another object of the present invention to provide a health food for preventing or ameliorating obesity or lipid-related metabolic diseases comprising an extract of black tea ethanolic aqueous solution as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating obesity or lipid-related metabolic diseases containing an extract of black tea ethanolic aqueous solution as an active ingredient.

In addition, the present invention provides a health food for preventing or ameliorating obesity or lipid-related metabolic diseases comprising an extract of black tea ethanolic aqueous solution as an active ingredient.

The black tea extract extracted with the ethanolic aqueous extract of the present invention has an increased gallic acid content as an active component and an indicator component of black tea as compared with the black tea extract extracted by the conventional hot water extraction and ethanol heating extraction method, The present invention relates to a pharmaceutical composition useful for the prevention or treatment of obesity, metabolic syndrome, diabetes and hyperlipemia, and is useful as an active ingredient of a health food, which exhibits excellent effects on weight and body fat reduction, fatty liver improvement, blood glucose control and cholesterol reduction in this obesity mouse model .

FIG. 1 is a graph showing a weight loss effect in a high fat diet-induced obese mouse model of a hot water extract of a cold water extract of hot water and a hot water extract of a black tea.
(* p < 0.05, ** p < 0.01 vs control)
FIG. 2 is a graph showing the body fat reduction effect in a high fat diet system of a cold water extract of hot water extract of black tea and a hot water extract in a mouse model of obesity.
FIG. 3 is a graph showing micro-CT photographs and fat areas comparing the body fat reduction effect in a high fat diet method of a cold water extract of black tea and a hot water extract in an obesity mouse model.
FIG. 4 is a graph showing the blood glucose lowering effect in the obesity mouse model of the high-fat diet of the ethanolic aqueous solution of black tea and the hot-water extract of black tea.
FIG. 5 is a graph showing the fatty liver improvement effect in the obesity mouse model of the high-fat diet of the ethanolic aqueous solution of black tea and the hot-water extract of black tea.
FIG. 6 is a graph showing a weight loss effect in an obesity mouse model of a high-fat diet of an ethanol aqueous solution of black tea extract and a heated extract of an aqueous ethanol solution.
FIG. 7 is a graph showing a body fat reduction effect in an obesity mouse model in which a high-fat diet of a black tea extract of a cold solution of an aqueous ethanol solution and an aqueous extract of an aqueous ethanol solution is used.
FIG. 8 is a graph showing blood glucose and blood cholesterol-lowering effects in a high-fat diet model of a black cohosh extract of ethanol aqueous solution and a heated extract of ethanol aqueous solution. FIG.
FIG. 9 is a graph showing the fatty liver improvement effect of the high-fat diet of the ethanolic aqueous solution extract of black tea and the ethanolic aqueous extract of black tea in an obesity mouse model.
FIG. 10 is a graph showing the inhibitory effect on the adipocyte differentiation of the ethanolic aqueous solution of black tea by the cold-dip extract. FIG.
11 is a graph showing an effect of inhibiting the accumulation of fat in mature adipocytes of the ethanolic aqueous solution of black coats extract of black tea.
FIG. 12 is a graph showing inhibitory effect on proliferation of lipid precursor cells and lipid differentiation of ethanol extract solution of black tea in black tea.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating obesity or lipid-related metabolic diseases comprising an extract of black tea ethanolic aqueous solution as an active ingredient.

The black tea may be cultivated or commercially available.

The black tea can be used for a variety of purposes such as a wagon, a tram, a cross, a bifurcation, a gold and a tolerance, a kangwon, a kinship, a battlefield, a black warrior, ), White tea, black tea, and black tea, but is not limited thereto.

The aqueous ethanol solution is preferably 30% to 70% ethanol aqueous solution, most preferably 50% ethanol aqueous solution, but is not limited thereto.

The metabolic diseases may be at least one selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis and lipid-related metabolic syndrome, but the present invention is not limited thereto.

In addition, the black tea ethanolic aqueous solution of the black tea extract preferably has a gallic acid content of 2 to 8% (w / w), but is not limited thereto.

In addition, it is preferable to reduce the body weight of the black tea ethanolic aqueous solution, more preferably to reduce body fat, and particularly to reduce body fat by decreasing abdomen or subcutaneous fat, but it is not limited thereto . Furthermore, it is preferable that the body fat reduction is a reduction in body fat caused by inhibition of lipid differentiation, and the lipid differentiation inhibition is suppressed not only by suppressing lipid-differentiated C / EBP? And PPAR? 2 genes but also by expressing the target genes FABP4, FAS and ACC It does not affect.

In addition, it is preferable that the body fat reduction is reduction of body fat through inhibition of proliferation of lipid precursor cells or adipocytes.

In addition, the black tea ethanolic aqueous solution of the black tea extract preferably improves the fatty liver and is preferably, but not limited to, a decrease in blood cholesterol or blood sugar.

In addition, the black tea ethanolate solution frozen extract of the present invention contains no adverse effects that cause metabolic diseases including insulin-resistant diabetes due to the degradation of PPARγ2 in adipocytes.

In addition, the black tea ethanol aqueous solution cold extract of the present invention is preferably, but not limited to, prepared by a method comprising the steps of:

1) mixing the black tea with an aqueous 30% to 70% ethanol solution at a ratio of 1: 4 to 1: 8;

2) extracting the mixture of step 1) for 60 to 80 hours; And

3) filtration of the extract of step 2), followed by concentration under reduced pressure.

In preparing the black tea extract of black tea ethanol aqueous solution of the present invention, the black tea of step 1) is preferably mixed with 50% ethanol aqueous solution, but not limited thereto.

In the production of the black tea extract of black tea ethanol aqueous solution of the present invention, shaking extraction, Soxhel extraction or reflux extraction is preferably used, but not limited thereto. It is preferable that the extraction solvent is added by 4 to 8 times the amount of the dried black tea, and it is most preferable to extract by adding 6 times. The extraction temperature is preferably 5 ° C to 35 ° C, and most preferably 10 to 30 ° C, but is not limited thereto. The extraction time is preferably 24 to 100 hours, more preferably 60 to 80 hours, most preferably 72 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 to 4 times, and most preferably, 3 times, but is not limited thereto.

In the preparation of the black tea extract of black tea ethanol aqueous solution of the present invention, the vacuum concentration of the vacuum concentrator or the vacuum rotary evaporator may be used for the concentration in step 3), but not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

In a specific example of the present invention, the inventors of the present invention conducted a study to determine the effect of the black tea extract of black tea ethanol aqueous solution as a composition for the prevention and treatment of obesity or lipid-related metabolic diseases in a black tea by ethanol concentration, extraction time, The gallic acid concentration of the obtained cold extract of the black tea ethanol aqueous solution obtained was measured by high performance liquid chromatography. As a result, it was found that 50% ethanol was dissolved in a black tea ethanol aqueous solution It was confirmed that the extract had a high content of gallic acid (see Tables 1 to 4).

In order to examine the effect of the black tea ethanol aqueous solution and the hot water extract of the present invention on the prevention and treatment of obesity or lipid-related metabolic diseases, the hot water extract of the black tea ethanol aqueous solution of the present invention was applied to a high fat The body weight, body fat, blood glucose and hepatic pathology were measured by oral administration to a diet-induced obese mouse model. The results showed that the high-fat diet had a weight loss of 23.5% , 100% of hot water extract of black tea decreased by 18.1%, that of black tea decreased by 13.2%, that of 100% of hot water extract of black tea decreased by 3.9%, that of hot water extract of black tea decreased by 18.1% Respectively. The body weight of the high fat diet dietary control group increased by about 10.24 g after 8 weeks of administration, but the weight of 300 ㎎ / ㎏ of 50% ethanol aqueous solution of black tea decreased by about 0.22 g, 23.5% decrease). The body weight of 100% hot water extract of 300 mg / kg of black tea was increased by about 1.94 g, and the weight loss effect was 18.1% lower than that of the control group (see FIG. 1 and Table 6).

In addition, the change in body weight was confirmed by the change in body fat rather than the change in the body weight or lean body mass, and the body fat of the high fat dietary diet control group increased by about 7.73% after 8 weeks, % Ethanolic water extract of 300 ㎎ / ㎏ was decreased by about 7.25%, which was superior to that of the control group. Body fat of 100% black tea extract (300 mg / kg) was reduced by about 3.18%, and body fat reduction was observed compared to the control group (see FIG. 2 and Table 7).

In addition, the abdominal fat and subcutaneous fat of the positive control group that consumed the high-fat diets for 8 weeks increased significantly compared to the normal group in the micro-CT photographs. However, abdominal fat and subcutaneous fat of mice administered with 50 ㎎% ethanol aqueous solution of black ethanol extract of 300 ㎎ / ㎏ were found to have abdominal fat and subcutaneous fat almost similar to normal group even though they consumed high fat diet. The abdominal fat and subcutaneous fat of the 50% ethanol solution of black tea 300 ㎎ / ㎏ were 512.4 ± 77.3 mm ³ and 125.9 ± 86.8 mm ³ , respectively. The abdominal fat of the high fat diet dietary control group was 3383.4 ± 249.8 mm ³ , 973.0 ± 191.6 mm ³ (442.2 ± 81.9 mm ^{3 for} abdominal fat, 77.9 ± 62.3 mm ^{3 for} subcutaneous fat), respectively (Fig. 3 and Table 8).

After 8 weeks, blood glucose level of 300 mg / kg of 50% ethanol aqueous solution of black tea was 77.97 ± 9.65 mg / dL, which was almost half of that of the control group (158.96 ± 20.06 mg / dL of the high fat diet dietary control group / dL) (see Fig. 4 and Table 9).

In addition, as a result of histopathological examination of the mice after 8 weeks, the fatty liver of 50% ethanol aqueous solution of cold solution extract of 300 ㎎ / kg of black tea was remarkably decreased compared with the positive control, and the fatty liver improvement effect was confirmed (see FIG.

In addition, in order to examine the effect of the black tea ethanol cold extract and the ethanol hot extract of the present invention on the prevention and treatment of obesity or lipid-related metabolic diseases, the black tea ethanol extract of the present invention and the ethanol- Body fat, blood glucose, cholesterol, and liver histopathology. The results showed that after 4 weeks of administration, the high-fat diet was 50% black tea compared to the positive control group. The weight loss of ㎎ / ㎏ was decreased by 22.1%, and the weight loss of the 50% extract of 300 ㎎ / ㎏ of cooked rice was 16.5%. After 8 weeks of administration, weight loss of black tea 50%, 300 ㎎ / ㎏ of alcohol extract was 23.9%, and that of 50% ethanol aqueous extract of 300 ㎎ / ㎏ was 19.5%. In addition, the body weight of the control group increased by about 1.68 g after 8 weeks, but the body weight of 300 ㎎ / ㎏ of the 50% ethanol solution of black tea decreased by about 11.47 g. Respectively. The body weight of 300 ㎎ / kg of the extract of black tea 50% ethanol aqueous solution was decreased by 9.01 g, and the weight loss effect was compared with the control group (see FIG. 6 and Table 10).

After 8 weeks, body fat of control group increased by 0.73% compared with that of control group, but body fat of 300 ㎎ / ㎏ of cold solution extract of 50% ethanol solution of black tea decreased by 11.63% Respectively. Body fat of the black tea 50% ethanol aqueous extract (300 mg / kg) was reduced by about 6.48%, thus confirming the effect of reducing body fat compared to the control group (see FIG. 7 and Table 11).

In addition, it was confirmed that the blood glucose level of the cold solution extract of black tea 50% ethanol aqueous solution of 300 mg / kg was 32.94 ± 11.23 mg / dL after 8 weeks, significantly lowering the blood glucose level compared to the control group (blood glucose level of the high fat dietary food control group: 86.03 ± 14.02 mg / ). The blood glucose level of 300 mg / kg of 50% ethanol extract of black tea was 61.94 ± 8.98 mg / dL, which was lower than that of the control group. However, it was confirmed that the 50% ethanol solution of black tea extract had better blood glucose lowering effect.

In addition, it was confirmed that cholesterol of 300 mg / kg of a 50% ethanol aqueous solution of black tea was 170.29 ± 16.67 mg / dL, which was significantly lower than that of the control group (cholesterol of high fat dietary food control group: 229.16 ± 11.44 mg / dL). The cholesterol of the black tea 50% was found to be 177.95 ± 10.39 mg / dL of 300 ㎎ / ㎏ of the alcoholic extract of the alcoholic beverages, and the cholesterol was reduced to the same degree as that of the black tea extract of 50% of the black tea (see FIG. 8 and Table 12).

In addition, as a result of histopathological examination of the mice after 8 weeks, the fatty liver of 50% black tea extract (300 mg / kg) was significantly lowered compared to the control group.

As a result of measuring the inhibitory effect on the adipocyte differentiation using 50% ethanol extract of black tea, fat formation was inhibited depending on the concentration of ethanol extract of 50% ethanol. This was consistent with the results obtained by extracting and quantifying stained Oil Red-O. In addition, it was found that the concentration of ethanol extract of black tea 50% ethanol was 0, 100, 200, 250, 300 and 400 ㎍ / ㎖, Respectively. In addition, the adipocyte treated with the 50% ethanolic cold extract was confirmed by RT-PCR. As a result, the fatty lipid fraction was inhibited by the 50% ethanol ethanolic cold extract of black tea, and C / EBPα and PPARγ2 mRNA levels remained constant and did not affect the expression of their target genes FABP4, FAS and ACC. In the case of 50 ㎍ / ㎖ ethanol extract of black tea, 50 ㎍ / ㎖ of oil, it was measured through Oil Red-O determination while maintaining normal level without inhibiting gene expression of PPARγ2 and C / EBPα, the master transcription factors required for differentiation of adipocytes into adipocytes The amount of one fat was reduced by 50-70% (see FIG. 10).

Further, as shown in FIG. 11, it was confirmed that the ethanolic cold extract of black tea 50% ethanol of 300 μg / ml reduced the fat of the cells. In addition, the effect of lowering the amount of fat already held by mature adipocytes to about 70% even after treatment with 100 μg / ml of 50% ethanolic cold extract of black tea was confirmed. This was consistent with the results obtained by extracting and quantifying stained Oil Red-O. In addition, the lipid differentiation inhibition was suppressed by 50% ethanolic cold extract of black tea, and the amount of mRNA of C / EBPα and PPARγ2 remained constant, and the target genes FABP4, FAS and ACC (Fig. 11).

Therefore, it can be seen that the 50% ethanol extract of black tea has a fat reduction effect but does not decrease the expression of PPARγ2 and C / EBPα, lipid differentiation transcription factors. Since the degradation of PPARγ2 causes various diabetic diseases such as insulin resistance, the 50% ethanolic cold extract of black tea suggests that PPARγ2 may not be accompanied by side effects caused by the dysfunction.

In addition, the cell proliferation inhibitory effect of lipid precursor cells and lipid differentiation using 50% ethanol extract of black tea was confirmed, and the cell proliferation process in the early stage of lipid differentiation was not completely compared with the control group. On the other hand, proliferation of progenitor cells was also inhibited by treatment of 50% ethanolic cold extract of black tea with 300 μg / ml of non-differentiated 3T3-L1 adipose precursor cells. This inhibited lipogenesis by inhibiting lipid differentiation and limiting the number of progenitor cells (see FIG. 12).

Therefore, the black tea ethanolic cold extract of the present invention enhances the content of gallic acid, which is an active ingredient and an indicator ingredient of black tea, and the high fat diet is superior to the obesity mouse model in decreasing body weight and body fat, improving liver fat, controlling blood glucose and decreasing cholesterol And can be effectively used as an active ingredient of a pharmaceutical composition useful for prevention or treatment of obesity, metabolic syndrome, diabetes and hyperlipemia, and health food.

The composition of the present invention can be administered orally or parenterally (e.g., applied or intravenously, subcutaneously, intraperitoneally), but oral administration is preferred. As the agent for parenteral administration, the external preparation such as powders, granules, tablets, capsules, sterilized aqueous solutions, liquid preparations, non-aqueous solutions, suspensions, emulsions, syrups, suppositories and aerosols, The composition may be formulated in the form of a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment, a pasta or a cataplasm. However, the present invention is not limited thereto. The composition for topical administration may be anhydrous or water-based, depending on the clinical formulation. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like. Solid form preparations for oral administration include powders, granules, tablets, capsules, soft capsules, and the like. Examples of liquid formulations for oral use include suspensions, solutions, emulsions, syrups and aerosols. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances and preservatives .

The composition may be prepared by adding black tea to the active ingredient in addition to one or more pharmaceutically acceptable carriers for administration. The pharmaceutically acceptable carrier may be a mixture of saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components. If necessary, an antioxidant, , And other conventional additives such as a bacteriostatic agent may be added. In addition, a diluent, a dispersant, a surfactant, a binder and a lubricant may be additionally added to formulate the composition for injectable use such as an aqueous solution, a suspension or an emulsion.

The pharmaceutically acceptable additives according to the present invention are preferably contained in an amount of 0.1 to 90 parts by weight based on the composition.

The preferred dosage of the composition of the present invention varies depending on the degree of absorption of the active ingredient in the body, the age of the patient, the sex, and the degree of obesity, but can be appropriately selected by those skilled in the art. However, for the desired effect, in the case of an oral administration agent, the composition of the present invention is generally administered to an adult at a dose of 0.0001 to 100 mg / kg per day, preferably 0.001 to 100 mg / kg per 1 kg of body weight per day It is good. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The composition for treating metabolic diseases of the present invention may contain at least one active ingredient which exhibits the same or similar function in addition to the black tea ethanolic cold extract.

In addition, the present invention provides a health food for preventing or ameliorating obesity or lipid-related metabolic diseases comprising an extract of black tea ethanolic aqueous solution as an active ingredient.

The black tea may be cultivated or commercially available.

The black tea can be used for a variety of purposes such as a wagon, a tram, a cross, a bifurcation, a gold and a tolerance, a kangwon, a kinship, a battlefield, a black warrior, ), White tea, black tea, and black tea, but is not limited thereto.

The aqueous ethanol solution is preferably 30% to 70% ethanol aqueous solution, most preferably 50% ethanol aqueous solution, but is not limited thereto.

The metabolic diseases may be at least one selected from the group consisting of fatty liver, type 2 diabetes, hyperlipidemia, cardiovascular disease, arteriosclerosis and lipid-related metabolic syndrome, but the present invention is not limited thereto.

In addition, the black tea ethanolic aqueous solution of the black tea extract preferably has a gallic acid content of 2 to 8% (w / w), but is not limited thereto.

In addition, it is preferable to reduce the body weight of the black tea ethanolic aqueous solution, more preferably to reduce body fat, and particularly to reduce body fat by decreasing abdomen or subcutaneous fat, but it is not limited thereto . Furthermore, it is preferable that the body fat reduction is a reduction in body fat caused by inhibition of lipid differentiation, and the lipid differentiation inhibition is suppressed not only by suppressing lipid-differentiated C / EBP? And PPAR? 2 genes but also by expressing the target genes FABP4, FAS and ACC It does not affect.

In addition, it is preferable that the body fat reduction is reduction of body fat through inhibition of proliferation of lipid precursor cells or adipocytes.

In addition, the black tea ethanolic aqueous solution of the black tea extract preferably improves the fatty liver and is preferably, but not limited to, a decrease in blood cholesterol or blood sugar.

In addition, the black tea ethanolate solution frozen extract of the present invention contains no adverse effects that cause metabolic diseases including insulin-resistant diabetes due to the degradation of PPARγ2 in adipocytes.

In addition, the black tea ethanol aqueous solution cold extract of the present invention is preferably, but not limited to, prepared by a method comprising the steps of:

1) mixing the black tea with an aqueous 30% to 70% ethanol solution at a ratio of 1: 4 to 1: 8;

2) extracting the mixture of step 1) for 60 to 80 hours; And

3) filtration of the extract of step 2), followed by concentration under reduced pressure.

In preparing the black tea extract of black tea ethanol aqueous solution of the present invention, the black tea of step 1) is preferably mixed with 50% ethanol aqueous solution, but not limited thereto.

In the production of the black tea extract of black tea ethanol aqueous solution of the present invention, shaking extraction, Soxhel extraction or reflux extraction is preferably used, but not limited thereto. It is preferable that the extraction solvent is added by 4 to 8 times the amount of the dried black tea, and it is most preferable to extract by adding 6 times. The extraction temperature is preferably 5 ° C to 35 ° C, and most preferably 10 to 30 ° C, but is not limited thereto. The extraction time is preferably 24 to 100 hours, more preferably 60 to 80 hours, most preferably 72 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 to 4 times, and most preferably, 3 times, but is not limited thereto.

In the preparation of the black tea extract of black tea ethanol aqueous solution of the present invention, the vacuum concentration of the vacuum concentrator or the vacuum rotary evaporator may be used for the concentration in step 3), but not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

Therefore, the black tea ethanolic cold extract of the present invention enhances the content of gallic acid, which is an active ingredient and an indicator ingredient of black tea, and the high fat diet is superior to the obesity mouse model in decreasing body weight and body fat, improving liver fat, controlling blood glucose and decreasing cholesterol And can be effectively used as an active ingredient of a pharmaceutical composition useful for prevention or treatment of obesity, metabolic syndrome, diabetes and hyperlipemia, and health food.

The black tea ethanolic cold extract of the present invention can be provided as a food composition by mixing with a pharmaceutically acceptable carrier. The black tea ethanolic cold extract extract or lysate is preferably contained in a weight ratio of 0.01 to 99.9% based on the whole composition, but is not limited thereto.

When the black tea ethanolic cold extract of the present invention is used as a food or beverage additive, the black tea extract may be used as it is, or may be used together with other food or food ingredients and suitably used according to a conventional method. The mixing amount of the black tea ethanolic cold extract may suitably be determined according to its intended use (prevention, health or therapeutic treatment). In the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the black tea ethanolic cold extract has no problem in terms of safety and can be taken for a long period of time. There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, drinks, tea, , Alcoholic beverages and vitamin complexes. The liquid component to be added in addition to the black tea when formulated into beverage is not limited but may include various flavors or natural carbohydrates such as ordinary beverages as an additional ingredient. Examples of such natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose and polysaccharides such as those commonly used in dextrin and cyclodextrins, , Sorbitol, and erythritol. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention. Natural flavors such as taumarin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (e.g., saccharin, aspartame, etc.) can be used as flavors other than those described above .

In another aspect, the food composition of the present invention may be formulated into various flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, , Protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. The food composition of the present invention may also contain pulp for the production of fruit and vegetable drinks. These components may be used singly or in combination, and the proportion of such additives is generally selected in the range of 0.001 to 50 parts by weight based on the total weight of the composition.

In another embodiment, the black tea ethanolic cold extract of the present invention may be a powder, a granule, a tablet, a capsule, a syrup, or a drink.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples.

EXAMPLES The following Examples, Experimental Examples and Preparation Examples are illustrative of the present invention, and the contents of the present invention are not limited to the following Examples, Experimental Examples and Production Examples.

< Example  1> Preparation of black tea ethanol extracts by solvent concentration

The first grade borrowed from Yunnan Province in China, the black car produced and processed in 2006 ( Camellia sinensis L.), and then the impurities were removed, and well-dried herbal medicine was used for the experiment. 20 g of black tea was extracted with 120 ml of 15%, 30%, 50%, 70%, 85% and 96% ethanol aqueous solution (v / v) at room temperature of 10 to 30 ° C for 72 hours, Concentrated under reduced pressure to 60 ° C, and then lyophilized to obtain 0.54 to 3.46 g of a black tea ethanol aqueous solution cold extract solution. The content of gallic acid in the thus obtained black tea ethanolic solution freeze-dried extract was measured by high performance liquid chromatography.

As a result, as shown in [Table 1], the higher the concentration of ethanol, the higher the content of gallic acid. However, in consideration of the production yield and gallic acid content, the concentration of the extraction solvent suitable for mass production and industrialization was 50% (V / v) (Table 1).

< Example  2> Solvent By drain  Preparation of black tea ethanol extract

The first grade borrowed from Yunnan Province in China, the black car produced and processed in 2006 ( Camellia sinensis L.), and then the impurities were removed, and well-dried herbal medicine was used for the experiment. 80 ml, 120 ml, 160 ml and 200 ml of a 50% ethanol aqueous solution (v / v) were added to 20 g of black tea, and the mixture was extracted at room temperature from 10 to 30 ° C for 72 hours. The mixture was filtered and concentrated under reduced pressure at 45 to 60 ° C , And lyophilized to obtain 1.54 to 3.46 g of a black tea ethanol extract. The gallic acid content of the obtained black tea ethanol extract was measured by high performance liquid chromatography.

As a result, as shown in [Table 2], the most appropriate extraction method was set as a method of extracting with a 6-fold aqueous ethanol solution (v / v) having the highest production yield and gallic acid content (Table 2).

< Example  3> Preparation of ethanol extract of black tea by extraction time

The first grade borrowed from Yunnan Province in China, the black car produced and processed in 2006 ( Camellia sinensis L.), and then the impurities were removed, and well-dried herbal medicine was used for the experiment. (20 g) was added with 120 ml of a 50% ethanol aqueous solution (v / v), and the mixture was extracted at 24 to 48 ° C for 24 hours, 48 hours, 72 hours and 96 hours at a temperature of 10 to 30 ° C. The mixture was filtered and concentrated under reduced pressure at 45 to 60 ° C. To obtain 2.90 ~ 3.46 g of a black tea ethanol extract. The gallic acid content of the obtained black tea ethanol extract was measured by high performance liquid chromatography.

As a result, as shown in Table 3, the most suitable extraction time was set to 72 hours as the extraction method with the highest production yield and gallic acid content (Table 3).

< Example  4> Preparation of black tea reflux (heating) extract

After extracting the solvent from the black tea extract with 50% ethanol aqueous solution and 100% hot water, the black tea extract was prepared to compare the gallic acid content of the extract.

The first grade borrowed from Yunnan Province in China, the black car produced and processed in 2006 ( Camellia sinensis L.), and then the impurities were removed, and well-dried herbal medicine was used for the experiment. 50 g of black tea was refluxed with 1,000 ml of 0% and 50% aqueous ethanol solution (v / v) for 3 hours, filtered, concentrated under reduced pressure at 45 to 60 ° C, and lyophilized to obtain black tea 8.00 g, 8.50 g. The gallic acid content of the obtained black tea extract was measured by high performance liquid chromatography.

As a result, as shown in Table 4, it was confirmed that the content of gallic acid was decreased by comparing the black tea ethanol aqueous solution extract with the black tea ethanol extract of the present invention (Table 4).

< Example  5> of black tea extract Gallan ( Gallic acid ) Concentration analysis

Performance analysis of high performance liquid chromatography (HPLC) was performed to analyze the gallic acid concentration of the black tea ethanol extract solution and the black tea ethanol extract obtained from Examples 1 to 4. Respectively.

The HPLC was carried out using a column of CAPCELL PAK MG (4.6 x 250 mm, 5 μm, Shiseido), the mobile phase (A) was 0.6% formic acid and the (B) was acetonitrile, . In addition, a UV 280 nm detector was used, and 10 μl of sample was injected at a flow rate of 0.8 ml / min at a temperature of 30 ° C.

HPLC Mobile phase  Condition minute Mobile phase A Mobile phase B 0 95 5 30 82 18 30.1 0 100 40 0 100 40.1 95 5 50 95 5

<5-1> Sample preparation

A standard solution (Stock standard solution) was prepared for the HPLC analysis. About 10.0 mg of the standard solution of Gallic acid was precisely weighed into a 10 ml volumetric flask, and 50% aqueous ethanol solution (v / v) was added thereto. The standard solution was diluted successively, and a working standard solution was prepared in a concentration range of 0.01 to 0.2 mg / ml.

<5-2> Preparation of Test Solution

Approximately 30 mg of the sample of the aqueous solution of the black tea ethanol aqueous solution was precisely weighed, placed in a 10 ml volumetric flask, and 50% aqueous ethanol solution (v / v) was added and the mixture was sonicated for 20 minutes. The solution was cooled and mixed with 50% aqueous ethanol solution (v / v). After shaking well, about 1 ml was transferred to a 1.5 ml micro tube (EP tube) and centrifuged at 13,000 rpm for 5 minutes The supernatant was used as the test solution.

<5-3> Quantitative test

The gallic acid content of the standard solution and the test solution prepared as in Example <5-1> and Example <5-2> was measured by the high performance liquid chromatography.

<5-4> Content calculation

The concentration Ct of the gallic acid in the test solution in the chromatogram of the test solution and the standard solution was calculated by the following formula (1).

[ Equation  One]

Content of gallic acid (%) = Ct / Wt x 1000 x P

Ct: concentration of gallic acid in sample solution (mg / mL)

Wt: sample weight (mg)

P: Purity of gallic acid standard (% / 100)

Aqueous solution of black tea ethanol Cold water  The yield of the extract and Galvanic  content Raw material amount Solvent selection Amount of solvent Extraction temperature Extraction time Results (g) yield(%) Gallic acid (%) 20g 96% EtOH 120 ml Room temperature 3 days 0.54 2.7 5.69 20g 85% EtOH 120 ml Room temperature 3 days 2.10 10.5 4.65 20g 70% EtOH 120 ml Room temperature 3 days 2.82 14.1 3.88 20g 50% EtOH 120 ml Room temperature 3 days 3.46 17.3 3.24 20g 30% EtOH 120 ml Room temperature 3 days 3.16 15.8 3.11 20g 15% EtOH 120 ml Room temperature 3 days 3.06 15.3 3.25

menstruum By drain  Solution of black tea ethanol Cold water  The yield of the extract and Galvanic  content Raw material amount Solvent selection Amount of solvent Extraction temperature Extraction time Results (g) yield(%) Gallic acid (%) 20g 50% EtOH 80 ml Room temperature 3 days 1.54 7.7 2.89 20g 50% EtOH 120 ml Room temperature 3 days 3.46 17.3 3.24 20g 50% EtOH 160 ml Room temperature 3 days 3.22 16.1 3.16 20g 50% EtOH 200 ml Room temperature 3 days 3.16 15.8 2.67

A black tea ethanol solution Cold water  Yield of extract Raw material amount Solvent selection Amount of solvent Extraction temperature Extraction time Results (g) yield(%) Gallic acid (%) 20g 50% EtOH 120 ml Room temperature 24 hours 3.16 15.8 3.43 20g 50% EtOH 120 ml Room temperature 48 hours 2.90 14.5 3.21 20g 50% EtOH 120 ml Room temperature 72 hours 3.46 17.3 3.24 20g 50% EtOH 120 ml Room temperature 96 hours 3.26 16.3 3.00

The yield of the black tea reflux (heating) extract and Galvanic  content Raw material amount Solvent selection Amount of solvent Extraction temperature Extraction time Results (g) yield(%) Gallic acid (%) 50g 50% EtOH 1,000 ml 95 ~ 100 ℃ 3 hours 8.00 16.0 2.04 50g 100% water 1,000 ml 95 ~ 100 ℃ 3 hours 8.50 17.0 2.98

Therefore, it was confirmed that the content of gallic acid in the black tea ethanolic aqueous solution extract obtained by using 50% ethanol aqueous solution at 6 times (v / v) for 72 hours at room temperature of 10 to 30 ° C was higher than that of 100% Respectively.

< Experimental Example  1> 50% ethanol solution of black tea Cold water  The extract and 100% Heat number  High-fat diet with extract High fat diet - induced obese mouse model The effect

The anti-obesity effect was measured in a high fat diet-induced obesity model using the 50% ethanolic cold extract of the black tea obtained in Example 1 and the 100% hot water extract of the black tea obtained in Example 4. The obese mouse model used in the experiment was as follows. Three weeks old male C57 / BL6 male mice (Korea BioLink, Korea) were fed with standard diet and water at 22 ~ 24 ℃ and humidity 60 ~ 80% for 1 week. ) For 7 weeks. Each animal was weighed and the top 70% was selected as the experimental animal and the bottom 30% was eliminated. Each of the above-mentioned upper 70% mice was divided into 8 groups according to body weight and body fat, and each drug was orally administered between 9:00 and 11:00 daily for 8 weeks.

<1-1> Weight change

The weight of the test animals in each arm was measured once a week and each result was expressed as weight (g) after 8 weeks (ie after the last drug administration) of the weight before the administration of the drug.

As a result, as shown in Fig. 1 and Table 6, in the case of the high-fat diet, the 50% ethanol solution of black tea, 300 mg / kg of the cold extract extract had a weight loss of 23.5% The extract of 300 ㎎ / ㎏ of extract decreased by 18.1%, the extract of 100 ㎎ / ㎏ of 50% ethanolic aqueous solution decreased by 13.2% and the concentration of 100 ㎎ / ㎏ of 100 ㎎ / ㎏ of black tea decreased by 3.9%.

In addition, as shown in [Table 6], the body weight of the high fat diet diet positive control group was increased by about 10.24 g after 8 weeks of the administration, but the body weight of the cold solution extract of black alcohol 50% ethanol aqueous solution 300 mg / kg was decreased by about 0.22 g (23.5% decrease) compared to the control group. The body weight of 100% hot water extract of black tea was increased by about 1.94 g to 18.1% of the control group (FIG. 1 and Table 6).

High fat diet model of obesity ( High fat diet - induced obese mouse model ) &Lt; / RTI & Heat number  The extract and aqueous ethanol solution Cold water  Weight loss effect of extract Experimental group Initial weight (g) Weight after 8 weeks (g) Early
% Change in contrast (%) Control group
% Change in contrast (%) It's normal. 27.68 + - 0.66 29.99 + - 0.86 + 8.3% - High fat dietary diet positive control 35.26 ± 1.02 45.49 + 1.43 + 29.1% - 100% black tea Hot water extract 100 mg / kg 35.07 ± 0.91 43.70 ± 1.55 + 24.4% -3.9% 100% black tea Hot water extract 300 mg / kg 35.31 占 1.21 37.25 + - 2.12 + 4.1% -18.1% Black tea 50% Ethanol aqueous solution Cooling extract 100 mg / kg 35.23 ± 1.18 39.47 ± 0.83 + 12.3% -13.2% Black tea 50% Ethanol aqueous solution Cooling extract 300 mg / kg 35.04 ± 1.03 34.81 ± 1.73 -1.0% -23.5%

<1-2> Changes in body fat

The Minispec LF90 live animal analyzer (Bruker, Germany) is capable of quickly and accurately analyzing the amount of fat, non-biphasic fluid, and body fluid in the body without dissecting or killing it with NMR equipment using nuclear magnetic resonance principle. The mice were weighed and weighed and the mice were placed in a Minispec LF90 instrument without any anesthetic or sedation, and the ratio of fat and lean body mass was measured.

As a result, as shown in Fig. 2 and [Table 7], it was confirmed that the change in body weight was due to a change in body fat rather than a change in the lean body mass.

In addition, as shown in [Table 7], body fat of the high fat dietary diet control group increased by about 7.73% after 8 weeks, but body fat of the 50% ethanol solution of black tea 300 ㎎ / kg decreased by about 7.25% The effect of reducing body fat was confirmed. Body fat of 100% black tea extract (300 mg / kg) was reduced by about 3.18%, thus confirming the effect of reducing body fat compared to the control group (FIG. 2 and Table 7).

High fat diet model of obesity ( High fat diet - induced obese mouse model ) &Lt; / RTI & Heat number  The extract and aqueous ethanol solution Cold water  Effect of body fat reduction of extract Experimental group Initial body fat (%) Body fat (%) after 8 weeks % Change from initial (% P) It's normal. 6.38 ± 0.76 5.91 + - 0.76 -0.47% P High fat dietary diet positive control 24.24 + 1.39 31.97 + 0.80 + 7.73% P 100% black tea Hot water extract 100 mg / kg 24.12 ± 1.23 28.50 ± 1.56 + 4.38% P 100% black tea Hot water extract 300 mg / kg 24.27 + - 1.26 21.08 ± 3.25 -3.18% P Black tea 50% Ethanol aqueous solution Cooling extract 100 mg / kg 24.31 + - 1.13 26.85 + 1.13 + 2.54% P Black tea 50% Ethanol aqueous solution Cooling extract 300 mg / kg 24.34 + 1.11 17.10 ± 2.09 -7.25% P

<1-3> Micro-tomography microtomoraphy or computed tomography ; Micro - CT ) analysis

After anesthetized by intraperitoneal administration of Ketamine (Huons, 1.5 ml / kg) and xylazine (Bayer, 0.5 ml / kg) to mice, in vivo Micro CT (Skyscan 1076, Bruker AXS, Germany). The volume of abdominal subcutaneous fat and visceral fat was measured using the SkyScan CT analyzer version 1.11 (Bruker ASX, Germany), which was reconstructed three-dimensionally and measured from the second lumbar spine to the fifth lumbar spine .

As a result, as shown in FIG. 3, the abdominal fat and subcutaneous fat of the positive control group consuming the high-fat diets for 8 weeks in the micro-CT photographs were increased more than the normal group. However, abdominal fat and subcutaneous fat of mice administered with 50 ㎎% ethanol aqueous solution of black tea extract of 300 ㎎ / ㎏ were found to have abdominal fat and subcutaneous fat almost similar to normal group even though they consumed high fat diet.

In addition, as shown in Table 8, the abdominal fat and subcutaneous fat of the 50% ethanol aqueous solution cold extract of black tea of 300 mg / kg were 512.4 ± 77.3 mm ³ and 125.9 ± 86.8 mm ³ , respectively, and the abdomen of the high fat dietary- Fat 3383.4 ± 249.8 mm ³ , subcutaneous fat 973.0 ± 191.6 mm ³ (442.2 ± 81.9 mm ^{3 for} abdominal fat and 77.9 ± 62.3 mm ^{3 for} subcutaneous fat) in the normal group (FIG. 3 and Table 8).

High fat diet model of obesity ( High fat diet - induced obese mouse model ) &Lt; / RTI & Heat number  The extract and aqueous ethanol solution Cold water  Extract Abdominal fat  And subcutaneous fat reduction effect Experimental group Abdominal fat (mm ³ ) Subcutaneous fat (mm ³ ) It's normal. 442.2 + 81.9 77.9 ± 62.3 High fat dietary diet positive control 3383.4 ± 249.8 973.0 ± 191.6 100% black tea Hot water extract 300 mg / kg 1945.3 ± 6.7 780.9 + - 403.6 Black tea 50% Ethanol aqueous solution Cooling extract 300 mg / kg 512.4 ± 77.3 125.9 ± 86.8

<1-4> Changes in blood sugar

1 ml of blood was collected from the abdominal vein of the mouse on the day of the autopsy for the hematology test, and blood coagulation was prevented using the blood anticoagulant EDTA-2K. Plasma was separated from the blood using a centrifuge, and the biochemical profile of the plasma was measured using a blood biochemical automatic analyzer (20i, KoneLab).

As a result, as shown in Fig. 4 and [Table 9], the blood glucose level of 300 mg / kg of 50% ethanol aqueous solution of black tea after 8 weeks was 77.97 ± 9.65 mg / dL, (High-fat dietary diet: 158.96 ± 20.06 mg / dL in the positive control group) (FIG. 4 and Table 9).

High fat diet model of obesity ( High fat diet - induced obese mouse model ) &Lt; / RTI & Heat number  The extract and aqueous ethanol solution Cold water  Effect of extract on blood glucose Experimental group Blood sugar (mg / dL) It's normal. 54.18 ± 3.17 High fat dietary diet positive control 158.96 ± 20.06 100% black tea Hot water extract 100 mg / kg 154.56 ± 19.33 100% black tea Hot water extract 300 mg / kg 87.23 + - 16.74 Black tea 50% Ethanol aqueous solution Cooling extract 100 mg / kg 113.29 + - 8.61 Black tea 50% Ethanol aqueous solution Cooling extract 300 mg / kg 77.97 + 9.65

Pathology examination between <1-5>

The liver of the mice was extracted and fixed in 10% neutral buffered formalin (Sigma, HT50-1-640) for 24 hours or more. The fixed tissue was treated with an automatic tissue processor (Vacuum tissue processor ASP300, Leica), and paraffin blocks were made with a foraging machine (Embedding center EG1150H, Leica). The paraffin blocks were cut into 4 ㎛ thickness with a microtome (RM2145, Leica) and stained with H & E with an automatic stainer (Autostainner XL, Leica). The stained tissue specimens were examined with an optical microscope (Axioskop 2, Zeiss).

As a result, as shown in Fig. 5, pathological examination of the mice after 8 weeks showed that the high fat diet of the fat solution of the cold solution extract of 300 ㎎ / kg of 50% ethanol solution of black tea was significantly lower than that of the positive control, (Fig. 5).

< Experimental Example  2> 50% ethanol solution of black tea Cold water  The high fat diet of 50% ethanol aqueous solution extract of black tea extract and obesity mouse model High fat diet - induced obese mouse model The effect

The anti-obesity effect was compared and measured in the high fat diet model using the 50% ethanolic cold extract of the black tea obtained in Example 1 and the 50% ethanol aqueous heating extract of black tea prepared in Example 4 . The obese mouse model used in the experiment was as follows. Three weeks old male C57 / BL6 male mice (Korea BioLink, Korea) were fed with standard diet and water at 22 ~ 24 ℃ and humidity 60 ~ 80% for 1 week. ) For 24 weeks. Each animal was weighed and the top 70% was selected as the experimental animal and the bottom 30% was eliminated. The selected top 70% mice were divided into groups based on body weight and body fat with 8 mice per group, and each drug was orally administered between 9:00 and 11:00 daily for 8 weeks.

<2-1> Weight change

The weight and body fat of the test animals were measured once a week, and the results were expressed as weight (g) after 8 weeks (ie after the last drug administration) of the weight before the administration of the drug.

As a result, as shown in Fig. 6 and [Table 10], the weight loss of the cold extract of black tea 50% ethanol solution of 300 mg / kg of black tea extract was 22.1% after 4 weeks of administration, compared with the positive control, and 50% ethanol The 300 ㎎ / ㎏ of aqueous heating extract showed a weight loss of 16.5%. After 8 weeks of administration, the body weight loss of 300 mg / kg of 50% ethanol aqueous solution of black tea decreased by 23.9%, and the weight loss of 300 mg / kg of 50% ethanol aqueous solution was 19.5%. In other words, the 50% ethanol solution of black tea extract was more effective than the 50% ethanol extract.

In addition, as shown in [Table 10], the body weight of the high fat dietary food control group increased by about 1.68 g after 8 weeks of the administration, but the weight of the cold water extract of black tea 50% ethanol aqueous solution 300 ㎎ / kg decreased by about 11.47 g And the effect of weight loss was confirmed. The body weight of 300 ㎎ / kg of the extract of black tea 50% ethanol aqueous solution was decreased by 9.01 g, and the weight loss effect compared to the control group was confirmed (FIGS. 6 and 10).

High fat diet model of obesity ( High fat diet - induced obese mouse model ) &Lt; / RTI &gt; &lt; RTI ID = 0.0 & Cold water  Weight loss effect of extracts and ethanol extract Experimental group Initial weight (g) Weight after 8 weeks (g) Early
% Change in contrast (%) Control group
% Change in contrast (%) It's normal. 36.23 + 1.71 36.18 ± 1.85 -0.23% - High fat dietary diet positive control 53.01 + - 0.73 54.68 ± 0.90 + 2.33% - Black tea 50% Ethanol aqueous solution Cooling extract 300 mg / kg 53.09 + - 0.71 41.63 ± 1.99 -21.84% -23.87% Black tea 50% Ethanol aqueous solution Heat extract 300 mg / kg 53.04 0.71 44.03 + - 1.06 -17.06% -19.48%

<2-2> Changes in body fat

The body fat of each group of mice was measured as in Example <1-2> above.

As a result, as shown in FIG. 7 and [Table 11], it was confirmed that the weight loss of the above <2-1> was mostly due to the decrease of body fat.

In addition, as shown in [Table 11], the body fat of the high fat diets diet control group increased by 0.73% compared with the control group after 8 weeks, while the body fat of the 50% ethanol solution of black tea 300 ㎎ / ㎏ was decreased by about 11.63% Which was superior to the control group. Body fat of the black tea 50% ethanol extract aqueous extract of 300 mg / kg was reduced by about 6.48%, thus confirming the effect of reducing body fat as compared with the control group (FIG. 7 and Table 11).

High fat diet model of obesity ( High fat diet - induced obese mouse model ) &Lt; / RTI &gt; &lt; RTI ID = 0.0 & Cold water  Reduction of body fat in extracts and ethanol extracts Experimental group Body fat (%) before administration Body fat (%) after 8 weeks Early
Contrast change rate (% P) It's normal. 10.60 ± 1.07 11.32 + - 0.94 + 0.72% P High fat dietary diet positive control 29.98 ± 1.08 30.71 + - 1.14 + 0.73% P Black tea 50% Ethanol aqueous solution Cooling extract 300 mg / kg 29.53 + - 0.83 17.91 + - 2.49 -11.63% P Black tea 50% Ethanol aqueous solution Heat extract 300 mg / kg 29.57 ± 0.79 23.09 + 1.83 -6.48% P

<2-3> Changes in blood sugar and blood cholesterol

Blood glucose and blood cholesterol were measured in the same manner as in Example <1-4>.

As a result, as shown in Fig. 8 and Table 12, it was confirmed that the blood glucose level of 300 mg / kg of the cold solution extract of 50% ethanol aqueous solution of black tea was 32.94 ± 11.23 mg / dL after 8 weeks, Blood glucose of feed control diet positive control: 86.03 ± 14.02 mg / dL). The blood glucose level of 300 mg / kg of 50% ethanol extract of black tea was 61.94 ± 8.98 mg / dL, which was lower than that of the control group. However, it was confirmed that the 50% ethanol solution of black tea extract had better blood glucose lowering effect.

In addition, it was confirmed that cholesterol of 300 mg / kg of a 50% ethanol aqueous solution of black tea was 170.29 ± 16.67 mg / dL, which was significantly lower than that of the control group (cholesterol of high fat dietary food control group: 229.16 ± 11.44 mg / dL). The cholesterol of the black tea 50% ethanol aqueous solution extract of 300 mg / kg was 177.95 ± 10.39 mg / dL, and it was confirmed that cholesterol was reduced to the same degree as that of the 50% ethanolic black tea extract solution (FIG. 8 and Table 12).

High fat diet model of obesity ( High fat diet - induced obese mouse model ) &Lt; / RTI &gt; &lt; RTI ID = 0.0 & Cold water  Extracts and ethanol extracts Coles Terror reduction effect Experimental group Blood sugar (mg / dL) Total cholesterol (mg / dL) It's normal. 17.73 + - 5.62 129.08 + - 4.07 High fat dietary diet positive control 86.03 + - 14.02 229.16 + - 11.44 Black tea 50% Ethanol aqueous solution Cooling extract 300 mg / kg 32.94 + - 11.23 170.29 + 16.67 Black tea 50% Ethanol aqueous solution Heat extract 300 mg / kg 61.94 + 8.98 177.95 ± 10.39

Pathology examination between <2-4>

The histopathological examination of the liver was carried out in the same manner as in Example <1-5>.

As a result, as shown in Fig. 9, pathological examination of the mice after 8 weeks showed that the high fat diet of the fat solution of the cold solution extract of 300 ㎎ / kg of 50% ethanol solution of black tea was significantly lower than that of the positive control, (Fig. 9).

<Experimental Example 3> Inhibition of adipocyte differentiation by ethanol extract of black tea ethanol

<3-1> Measurement of triglyceride in adipocytes

The effect of inhibiting adipocyte differentiation was measured using the 50% ethanolic cold extract of black tea obtained in Example 1 above.

Specifically, 3T3-L1 adipose precursor cells used in the experiment were divided into 5 x 10 ³ cells / cm ² and cultured for 4 days. When the number of cells was increased and the growth was completely inhibited by contact inhibition, the differentiated hormone was treated for 6 days as follows.

0 day: 10% fetal bovine serum (FBS), 5 μg / ml insulin, 2 μM Dexamethasone (Dex), 0.5 mM isobutylmethylxanthine (IBMX)

2 days: 10% FBS, 5 [mu] g / ml Insulin,

4 days: 10% FBS,

6 days: Observation of differentiation and

6-10 days: At the end of differentiation, mature adipocytes were cultured in medium supplemented with 10% FBS for 4 days. The 50% ethanol extract of black tea was dissolved in the medium to a concentration of 20 mg / ml, filtered through a 0.2 μM filter and diluted to an appropriate concentration through serial dilution. 3T3-L1, an undifferentiated lipid precursor cell, was treated with adipocyte differentiation inducing hormone (0.5 mM 1-isobutyl-3-methylxanthine (IBMX), 2 M dexamethasone (Dex) and 5 g / ml insulin (Kirkland et al., 1990). At this time, cell differentiation was observed by treating the 50% ethanol extract of black tea with 0, 10, 25, 50, 100 and 200 ㎍ / ml. On day 6 of adipocyte differentiation, the medium was removed, washed twice with phosphate buffered saline (PBS), and fixed with 10% formaldehyde for 1 hour at room temperature. The neutral lipid staining was performed by dissolving Oil Red O (Sigma # O0625) in 0.5% isopropanol and diluting it in distilled water at a ratio of 3: 2. The immersed 3T3-L1 cells were stained with the Oil Red O solution to a sufficient extent to immerse them in the cells for 30 minutes at 37 ° C. Dyed Oil Red O was extracted again with isopropanol and quantified by measuring the absorbance at 510 nm wavelength.

As a result, as shown in Fig. 10, it was confirmed that fat formation was inhibited in dependence on the concentration of 50% ethanol ethanolic cold extract (Fig. 10A). This was in agreement with the results obtained by extracting and quantifying stained Oil Red-O (FIG. 10B). In addition, it was found that the 50% ethanol solution of black tea extract of black tea of 200 ~ 300 ㎍ / ㎖ was found to be sufficient for the inhibition of adipocyte differentiation by treatment with 0, 100, 200, 250, 300 and 400 ㎍ / (Fig. 10C).

<3-2> Measurement of lipid-differentiated transcription factor

After the differentiation, total RNA was isolated using RNeasy spin column (Qiagen, Chatsworth, CA, Cat. No. 74106). The separated RNA (2 ug) was reverse-transcribed for 1 hour at 37 ° C using M-MLV reverse transcriptase (Promega, Madison WI, Cat. No. M170B), dNTP and hexamer random primers, 20 ng) was quantitated into the ABI 7000 Real-Time PCR System (instrument) using a Power SYBR Green PCR Master Mix (Applied Biosystems, Cat. No. 4367659). The quantitative PCR is carried out by the following steps: 1) 50 ° C for 2 min, 2) 95 ° C for 10 min, 3) 95 ° C for 15 sec, 4) 60 ° C for 1 min and step 5) ) Was repeated 40 times, and the primer sequences used were (F): Forward primer and (R): Reverse primer).

C / EBP? (F) 5'-AGCCAAGAAGTCGGTGGACA-3 '(SEQ ID NO: 1),

C / EBP? (R) 5'-CAGCCTAGAGATCCAGCGACC-3 (SEQ ID NO: 2);

PPAR? 2 (F) 5'-CTGATGCACTGCCTATGAGCA-3 '(SEQ ID NO: 3),

PPAR? 2 (R) 5'-ATGCGAGTGGTCTTCCATCAC-3 '(SEQ ID NO: 4);

ACC (F) 5'-TAACAGAATCGACACTGGCTGGCT-3 '(SEQ ID NO: 5),

ACC (R) 5'-ATGCTGTTCCTCAGGCTCACATCT-3 '(SEQ ID NO: 6);

FAS (F) 5'-TGCTCCCAGCTGCAGGC-3 '(SEQ ID NO: 7),

FAS (R) 5'-GCCCCGTAGCTCTGGGTGTA-3 '(SEQ ID NO: 8);

FABP4 (F) 5'-GATGAAATCACCGCAGACGACA-3 '(SEQ ID NO: 9),

FABP4 (R) 5'-ATTGTGGTCGACTTTCCATCCC-3 '(SEQ ID NO: 10);

18s rRNA (F) 5'-ACCGCAGCTAGGAATAATGGAATA-3 '(SEQ ID NO: 11),

18s rRNA (R) 5'-CTTTCGCTCTGGTCCGTCTT-3 '(SEQ ID NO: 12).

The cells of Experimental Example <3-1> were subjected to RT-PCR to confirm the change of lipoprotein transcription factor. Expression of C / EBPα and PPARγ2, which are known as master transcription factors in adipocyte differentiation, is important.

As a result, as shown in FIG. 10, although the lipolysis was suppressed by the 50% ethanolic cold extract of black tea, the amounts of mRNAs of C / EBPα and PPARγ2 remained constant, and the expression of these target genes FABP4, FAS and ACC (Fig. 10D). In the case of 50 ㎍ / ㎖ ethanol extract of black tea, 50 ㎍ / ㎖ of oil, it is measured through Oil Red-O determination while maintaining the normal level without inhibiting gene expression of PPARγ2 and C / EBPα, the master transcription factors required for differentiation of adipocytes into adipocytes It was confirmed that the amount of one fat was reduced by 50-70%.

< Experimental Example  4> black tea ethanol Cold water  Identification of mature adipocyte inhibitory effect by extracts

<4-1> Measurement of triglyceride in adipocytes

In order to confirm mature adipocyte inhibitory effect using the 50% ethanolic black tea extract obtained in Example 1, mature 3T3-L1 adipocytes were injected with 100, 200, 300, 500, 750 and 1000 Ug / ml, and the cells were stained with Oil Red-O for 4 days.

As a result, as shown in Fig. 11, it was confirmed that a cold 50% ethanol ethanol extract of 300 μg / ml of black tea reduced the fat of the cells (FIG. 11A). In addition, the effect of lowering the amount of fat already held by mature adipocytes to about 70% even after treatment with 100 μg / ml of 50% ethanolic cold extract of black tea was confirmed. This was confirmed to be consistent with the results obtained by extracting and quantifying stained Oil Red-O (FIG. 11B).

<4-2> Measurement of lipid differentiation transcription factor

The cells of Experimental Example <4-1> were examined for the change of lipoprotein transcription factor by the RT-PCR method of the above Example <3-2>.

As a result, as shown in Fig. 11, the amount of mRNA of C / EBP? And PPAR? 2 remained constant despite inhibition of lipidation by the 50% ethanolic cold extract of black tea, and the expression of these target genes FABP4, FAS and ACC (Fig. 11C).

Therefore, 50% ethanol extract of black tea showed fat reduction effect but it did not decrease the expression of PPARγ2 and C / EBPα, which are lipogenic transcription factors. Since the degradation of PPARγ2 causes various metabolic diseases such as insulin resistance, the 50% ethanolic cold extract of black tea suggests that PPARγ2 may not be accompanied by the side effects caused by the dysfunction.

< Experimental Example  5> black tea 50% ethanol Cold water  Extracts inhibited proliferation of lipid precursor cells and lipid differentiation

In order to confirm the cell proliferation inhibitory effect upon lipid precursor cells and lipid differentiation using the 50% ethanol coagulant extract obtained in Example 1, 3T3-L1 cells were used to dissolve 50% ethanolic black tea extract Was treated at a concentration of 300 / / ml to observe the morphology of the cells with an optical microscope and confirm the number of living cells.

As a result, as shown in Fig. 12, the cell proliferation process in the early stage of fat differentiation did not appear completely in comparison with the control group (Fig. 12A and Fig. 12B). On the other hand, it was confirmed that proliferation of progenitor cells was also inhibited by treatment of 50% ethanolic cold extract of black tea with 300 μg / ml of non-differentiated 3T3-L1 adipose precursor cells (FIG. 12C).

Therefore, it was confirmed that lipogenesis is inhibited by inhibiting lipid differentiation and limiting the number of progenitor cells.

< Manufacturing example  1> Preparation of injection preparation

100 mg of the black tea ethanolic aqueous solution of black tea extract of the present invention

Sodium metabisulphite 3.0 mg

Methyl paraben 0.8 mg

0.1 mg of propylparaben

14 mg of sodium chloride

Sterile sterilized water for injection

The above components were mixed and made up to 2 ml in a usual manner, filled in a 2 ml ampoule and sterilized to prepare an injection.

< Manufacturing example  2> Preparation of tablets

200 mg of the black tea extract aqueous solution of black tea of the present invention

Lactose 100 mg

50 mg of microcrystalline cellulose

Crospovidone 20 mg

Magnesium stearate qs

The above components were mixed and tablets were prepared by tableting according to a conventional method for producing tablets.

Preparation Example 3 Preparation of capsules

100 mg of the black tea ethanolic aqueous solution of black tea extract of the present invention

Lactose 50 mg

20 mg of low-substituted hydroxypropylcellulose

5 mg of silicon dioxide

2 mg of talc

Magnesium stearate qs

The above components were mixed and filled in gelatin capsules according to the conventional preparation method of capsules to prepare capsules.

< Formulation example  4> Preparation of liquid agent

1000 mg of the black tea extract aqueous solution of the present invention

20 g of white sugar

3 g of anhydrous citric acid

Fructose 20 g

Lemon incense quantity

Purified water was added to 100 ml

The above components were mixed according to a conventional method for preparing a liquid preparation, filled in 100 ml of a brown bottle, and sterilized to prepare a liquid preparation.

&Lt; 110 > Dong-A Pharmaceutical Co., Ltd. <120> Pharmaceutical composition or health food containing black tea          ethanol solution extracts which are effective for reducing body          weight and body fats or preventing or treating lipids related          metabolic disease <130> 12P-12-032 <160> 12 <170> Kopatentin 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> C / EBP alpha Forward primer <400> 1 agccaagaag tcggtggaca 20 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> C / EBP alpha Reverse primer <400> 2 cagcctagag atccagcgac c 21 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PPAR gamma 2 Forward primer <400> 3 ctgatgcact gcctatgagc a 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PPAR gamma 2 Reverse primer <400> 4 atgcgagtgg tcttccatca c 21 <210> 5 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> ACC Forward primer <400> 5 taacagaatc gacactggct ggct 24 <210> 6 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> ACC Reverse primer <400> 6 atgctgttcc tcaggctcac atct 24 <210> 7 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> FAS Forward primer <400> 7 tgctcccagc tgcaggc 17 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> FAS reverse primer <400> 8 gccccgtagc tctgggtgta 20 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> FABP4 Forward primer <400> 9 gatgaaatca ccgcagacga ca 22 <210> 10 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> FABP4 Reverse primer <400> 10 attgtggtcg actttccatc cc 22 <210> 11 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> 18S mRNA Forward primer <400> 11 accgcagcta ggaataatgg aata 24 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 18S mRNA reverse primer <400> 12 ctttcgctct ggtccgtctt 20

Claims (34)

A pharmaceutical composition for the prevention or treatment of obesity comprising an extract of black tea ethanolic aqueous solution as an active ingredient.
The method according to claim 1, wherein the black tea is selected from the group consisting of a bottle, a tank, a chariot, a chariot, a gold and a tolerance, a kang, a gold leaf, Wherein the composition is selected from the group consisting of open-air bottles, white tea, black tea, and black tea.
The pharmaceutical composition for preventing or treating obesity according to claim 1, wherein the aqueous ethanol solution is a 30% to 70% (v / v) aqueous ethanol solution.
2. The method according to claim 1, wherein the black tea ethanolate aqueous coolant extract
1) mixing the black tea with an aqueous solution of 30% to 70% (v / v) of ethanol in a ratio of 1: 4 to 1: 8 (w: v);
2) extracting the mixture of step 1) for 60 to 80 hours; And
3) The pharmaceutical composition for preventing or treating obesity, which comprises the step of filtering and then concentrating the extract of step 2) under reduced pressure.
2. The method according to claim 1, wherein the black tea ethanolate aqueous coolant extract
1) mixing the black tea with a 50% (v / v) aqueous ethanol solution at a ratio of 1: 6 (w: v);
2) extracting the mixture of step 1) at 10 to 30 DEG C for 72 hours; And
3) The pharmaceutical composition for preventing or treating obesity, which comprises the step of filtering and then concentrating the extract of step 2) under reduced pressure.
delete The pharmaceutical composition for preventing or treating obesity according to claim 1, wherein the black tea ethanolic solution frozen extract has a gallic acid content of 2 to 8% (w / w).
The pharmaceutical composition for preventing or treating obesity according to any one of claims 1 to 3, wherein the black tea ethanolate aqueous solution is reduced in body weight.
The pharmaceutical composition for preventing or treating obesity according to claim 1, wherein the black tea ethanol-aqueous solution cold extract extract reduces body fat.
10. The pharmaceutical composition for preventing or treating obesity according to claim 9, wherein the decrease in body fat is a decrease in abdomen or subcutaneous fat.
10. The pharmaceutical composition for the prevention or treatment of obesity according to claim 9, wherein the body fat reduction is inhibited by lipidation to cause body fat loss.
12. The method for preventing or treating obesity according to claim 11, wherein the lipid differentiation inhibition does not affect the expression of the target genes FABP4, FAS and ACC of the lipid differentiation transcription factors C / EBPa and PPAR? A pharmaceutical composition.
The pharmaceutical composition for preventing or treating obesity according to claim 9, wherein the body fat reduction is inhibited by proliferation of lipid precursor cells or adipocytes to reduce body fat.
[Claim 2] The pharmaceutical composition for preventing or treating obesity according to claim 1, wherein the black tea ethanolic solution frozen extract of the black tea improves fatty liver.
The pharmaceutical composition for preventing or treating obesity according to claim 1, wherein the black tea ethanolate solution cold extract extract is reduced in blood cholesterol.
The pharmaceutical composition for prevention or treatment of obesity according to claim 1, wherein the black tea ethanolate solution cold extract extract is reduced in blood sugar.
[Claim 2] The pharmaceutical composition for preventing or treating obesity according to claim 1, wherein the black tea ethanolate solution freeze-dried extract has no side effects that cause metabolic diseases including insulin-resistant diabetes caused by a decrease in function of PPARγ2 in adipocytes.
A health functional food for prevention or improvement of obesity comprising an extract of black tea ethanol aqueous solution of black tea as an active ingredient.
19. The method according to claim 18, wherein the black tea is selected from the group consisting of a bottle, a tank, a chariot, a chariot, a gold and a tolerance, a kang, a gold leaf, Wherein the composition is selected from the group consisting of open-air bottles, white tea, black tea, and black tea.
19. The health functional food for preventing or improving obesity according to claim 18, wherein the aqueous ethanol solution is an aqueous ethanol solution of 30% to 70% (v / v).
19. The method according to claim 18, wherein the black tea ethanol-aqueous solution cool-
1) mixing the black tea with an aqueous solution of 30% to 70% (v / v) of ethanol in a ratio of 1: 4 to 1: 8 (w: v);
2) extracting the mixture of step 1) for 60 to 80 hours; And
3) filtering the extract of step 2), and concentrating the filtrate under reduced pressure, thereby producing a health functional food for preventing or improving obesity.
19. The method according to claim 18, wherein the black tea ethanol-aqueous solution cool-
1) mixing the black tea with a 50% (v / v) aqueous ethanol solution at a ratio of 1: 6 (w: v);
2) extracting the mixture of step 1) at 10 to 30 DEG C for 72 hours; And
3) filtering the extract of step 2), and concentrating the filtrate under reduced pressure, thereby producing a health functional food for preventing or improving obesity.
delete [19] The health functional food for preventing or improving obesity according to claim 18, wherein the black tea ethanolic solution freeze-dried extract has a gallic acid content of 2 to 8% (w / w).
[19] The health functional food for preventing or improving obesity according to claim 18, wherein the black tea ethanol-aqueous solution cold extract extract is reduced in body weight.
19. The health functional food for preventing or ameliorating obesity according to claim 18, wherein the black tea ethanol-aqueous solution cold extract extract reduces body fat.
27. The health functional food for preventing or improving obesity according to claim 26, wherein the decrease in body fat is a decrease in abdomen or subcutaneous fat.
27. The health functional food for preventing or ameliorating obesity according to claim 26, wherein the reduction in body fat is inhibited by lipidation to cause a decrease in body fat.
29. The method according to claim 28, wherein the lipid differentiation inhibition does not inhibit the expression of the target genes FABP4, FAS and ACC of the lipid differentiation transcription factors C / EBP &lt; alpha & Health functional foods.
27. The health functional food for preventing or ameliorating obesity according to claim 26, wherein the body fat reduction is inhibited by proliferation of lipid precursor cells or adipocytes to reduce body fat.
19. The health functional food for preventing or ameliorating obesity according to claim 18, wherein the black tea extract of ethanol solution of black tea is improved in fatty liver.
The health functional food for preventing or ameliorating obesity according to claim 18, wherein the black tea extract of black tea ethanol aqueous solution is caused by a decrease in blood cholesterol.
The health functional food for preventing or ameliorating obesity according to claim 18, characterized in that the black tea extract of black tea ethanol aqueous solution is caused by a decrease in blood glucose.
19. The health functional food for obesity prevention or improvement according to claim 18, wherein the black tea ethanolate solution frozen extract of the black tea has no side effects that cause metabolic diseases including insulin resistance diabetes due to a decrease in function of PPAR? 2 in adipocytes.

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