KR20100002658A - Extract of polygonum cuspidatum, compositions and functional food for prevention and treatment of obesity - Google Patents

Abstract

PURPOSE: A pharmaceutical composition for preventing and treating obesity using Polygoni Rhizoma extract and its ethylacetate layer fraction and butanol fraction is provided to suppress activation of adipose and adipose absorption into intestine tract. CONSTITUTION: A pharmaceutical composition for preventing or treating obesity contains Polygoni Rhizoma extract as an active ingredient. The composition also contains ethylacetate fraction of Polygoni Rhizoma extract obtained through normal hexane, ethylacetate, butanol, and water.

Description

Extract of Polygonum cuspidatum, Compositions and functional food for prevention and treatment of obesity

The present invention relates to a pharmaceutical composition for the prevention or treatment of obesity, and a dietary supplement for improvement, a hyperlipidemia or a metabolic syndrome prevention and treatment, and a dietary supplement containing the butanol layer fraction of K. koji extract as an active ingredient. .

Obesity refers to a state in which excess energy is accumulated as fat due to high energy intake compared to energy consumption. According to the 2005 National Health and Nutrition Survey, 31.8% of adults over 20 years of age in Korea were obese. More than doubled over a decade (Ministry of Health & Welfare. 2006. The third Korea National Health and Nutrition Examination Survey). According to this increase, the socioeconomic loss caused by obesity is increasing from 1.7 trillion won in 2001, and the government's comprehensive national health promotion plan aims to maintain the adult obesity rate at 30% by 2010. (Ministry of Health and Welfare. 2005. Comprehensive National Health Promotion Plan, p 61-64). Obesity is not only a problem of its own, but also can cause diseases such as cardiovascular disease, diabetes, respiratory disease, osteoarthritis, etc., thus attracting much attention to obesity worldwide (Antipatis VJ et al., 2001, Obesity as a global problem.In International textbook of obesity . Per Bjorntorp, ed. John Wiley & Sons Ltd, Chichester, UK. p3-22), the World Health Organization (WHO) treats obesity as a global nutrition problem and recognizes it as a disease to be treated, not as a simple risk factor to health (World Health Organization. 1998. Obesity: Preventing and managing the global epidemic.Report of WHO Consultation on Obesity, Geneva). Obesity is due to a combination of excessive energy consumption, lack of exercise, as well as neurological secretion factors, drug causes, genetic factors, etc. As a way to improve obesity, diet, exercise, behavioral therapy or drug treatment is used.

Recently, many studies for the development of obesity treatments have been conducted, and one of these studies is a pancreatic lipase inhibitor. Pancreatic lipase is a key enzyme that breaks down triglycerides into 2-monoacylglycerol and fatty acids (Bitou N. et al., 1999, Lipid 34: 441-445). There is tetrahydrolipstatin (Orlistat). Orlistat, currently available as a medicinal product, is known to be most potent, inhibiting about 30% of the fat consumed (Drent M. et al., 1995, Int . J. Obesity . 19: 221-226). Despite these effects, however, there are known side effects such as gastrointestinal disorders, hypersensitivity, bile secretion disorders, and fat-soluble vitamin absorption inhibition (Peter C. et al., 2001, Br . J. Clin. Pharmacol . 51: 135- 141). In addition, FDA's weight loss treatment regulations require long-term anti-obesity drug effects to be reduced by 5% of body weight before drug administration, and the weight loss effect must be maintained for at least 12 months. However, Orlistat loses only 3 to 5% of body weight, even if administered for 2 years, and has side effects that interfere with fat-soluble vitamins A, D and E absorption. It was launched in 1998 and generated sales of $ 96,300 in 2001. Since then, the company has been losing sales due to the lack of side effects and effects. Therefore, recently, research for the development of pancreatic lipase inhibitors from foods and natural products with no side effects (Yamamoto M. et al., 2000, Int . J. Obesity . 24: 758-764).

The results of the development of anti-obesity materials using natural products show that dietary fibers such as hydroxycitric acid (HCA), Olibra, chicory, and inulin are involved in appetite suppression according to the mechanism of action. Substances include chitosan and flavonoids. Examples of substances that induce heat generation and inhibit fat accumulation include capsaisin of red pepper, catechin of green tea, and retinoic acid, and L-carnitine, conjugated linoleic acid (CLA), which regulates lipid metabolism, and milk calcium and related proteins. It is. Herbal medicines such as fruit, yangha, deficiency, green tea, pine needles, hwahwa, cheongung, and filthy milk have been reported to be effective for weight control (Kim MH, 2004, Korean J. Heath Psychol. 9: 493-509, Reddy P. et al., 1998, Formulary 33: 943-959, Burns AA et al., 2002, Eur . J. Clin . Nutr . 56: 368-377, Delzenne NM et al., 2005, Br . J. Nutr . 1:.. 157-161, Zacour AC et al, 1992, J. Nutr Sci Vitamilol 38:..... 609-613, Griffiths DW et al, 1986, Adv Exp Med Biol 1999:. 509-516) . Therefore, consumers are less interested in such side effects, and on the contrary, they are more interested in natural anti-obesity drugs, which are excellent in efficacy, and to meet these demands, development of safe and effective natural medicines is desired.

Accordingly, the present applicant has continued research on the prevention and treatment of obesity, and the present invention suppresses the activity of pancreatic lipase, which is an important enzyme involved in energy intake and double fat absorption, which are one of the main causes of obesity. Providing butanol-layered fraction of S. aeruginosa extract, which has been proven to inhibit weight gain and reduce weight in obesity induced by high fat diet and short-term fat absorption using fat emulsion, provides a pharmaceutical composition for preventing and treating obesity It aims to do it.

In addition, an object of the present invention is to provide a hyperlipidemia or metabolic disease pharmaceutical composition and a functional food containing the butanol layer fraction of E. coli muscle extract of the present invention as an active ingredient.

In the extract of Escherichia coli obtained by extracting Escherichia coli with an alcohol or an aqueous solution of alcohol, and filtering the extracted extract and concentrating under reduced pressure, a pharmaceutical composition for preventing or treating obesity containing the Escherichia coli extract as an active ingredient and preventing and improving obesity It is characterized by improving the health functional food.

In addition, the above-mentioned extract of ephedra roots in the order of normal hexane, ethyl acetate, butanol, water to prevent obesity containing the ethyl acetate layer fraction and butanol layer fraction of the E. It is characterized by improving the pharmaceutical composition for treatment and health functional foods for the prevention and improvement of obesity.

E. coli extract and acetate layer fraction, butanol layer powder of the extract according to the present invention has the effect of inhibiting lipase activity, inhibit the absorption of intestinal fat. In addition, anti-obesity experiments induced by high fat diet have weight gain suppression and weight loss effects.

Accordingly, the butanol layer fraction of Escherichia coli extract according to the present invention can be applied to pharmaceutical compositions and functional foods for the prevention and treatment of obesity.

In addition, since anti-obesity experiments induced by high-fat diet have demonstrated the effect of lowering total cholesterol, triglycerides and LDL cholesterol in blood, it may be applied to pharmaceutical compositions and functional foods for the prevention and treatment of hyperlipidemia.

In addition, since obesity is an important cause of metabolic syndrome, butanol layer fraction according to the present invention will be applicable to pharmaceutical compositions and functional foods for the prevention and treatment of metabolic syndrome.

The present invention provides a pharmaceutical composition for the prevention or treatment of obesity, and a dietary supplement for improvement or prevention of hyperlipidemia or metabolic syndrome, which contain the extract of E. coli, Echilacetate layer of E. coli extract and butanol layer of E. coli extract as an active ingredient. And it is characterized in that the therapeutic pharmaceutical composition and health functional food.

Hogon ( Polygonum cuspidatum Sieb . et Zucc . = Reynotria japonica Houtt.) Is a perennial herb belonging to Polygonaceae. The root stem is thick and its stem is about 1.5 m high. Leaves are alternate, egg-shaped, 6-12 cm long, with sharp ends, petioles long. Flowers bloom in males and females in June-August. Fruits are aquatic, triangular, egg-shaped, with a dark brown luster. It grows in valleys throughout the country and is distributed in Japan, China, and Taiwan (Bae Gi-hwan, Korean medicinal plants, Kyohaksa, 2003, 89). Root roots and roots are called Hojang or Hojanggeun (Poly 根, Polygoni Rhizoma = Reynoutriae Rhizoma) and are harvested in the 本草綱目 木 部 中 品. In Korea, it has been used for the treatment of palliative, diuretic, purulent dermatitis, cystitis, and anticancer, and in oriental medicine, it is used for the wind moisturizing, pore, cultivation, Jinhae and jingyeong. , 1997, p. 140; Hell Mark, 1975, Journal of Pharmacognosy, 6: 1-4). The pharmacological action of Kho-Geun-Geun included lipid metabolism inhibition (Masaki H., et al., 1995, Biol . Pharm . Bull . 18: 162-166, 1995), protein tyrosine kinase inhibitory activity (Jayatilake GS, et al., 1993, J. Nat . Prod . 56: 1805-1810), mutagenicity inhibition (Su et al., 1995, Mutat. Res . 329: 205-212), and the like. The major components of eosinophilia are stilbene, anthraquinone, and glycosides such as polydatin, resveratrol, emodin, physcion, and chrysophanoic acid (Pan et al., 2000, Zhong). Yao Cai , 23: 56-58; Zhang, 1999, Tianjin Yi Yao , 11: 13-14).

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

I. Root Muscle Extract  Method for preparing butanol layer fraction

Hos used in the experiment [ Polygonum cuspidatum Sieb . et Zucc. = Reynotria japonica Houtt .; Polygonaceae] was supplied from Yunnan Province, China, and the specimen (no. KIOM-POCU1) was stored in the herbarium of the Korean Institute of Oriental Medicine.

1) Extraction step

Ethanol was added to the dry and dried Hojang-geun, repeatedly extracted at room temperature in an extraction container, and concentrated under reduced pressure at 40 ° C. to obtain an ethanol extract.

2) concentration step

 The extract is filtered and concentrated under reduced pressure. At this time, the temperature should be maintained at 40-45 ° C. or lower during concentration to prevent decomposition and hydrolysis of the components.

3) Escherichia coli extract system separation step

Suspension of ethanol extract prepared by the above method in suspension in distilled water and then sequentially separated into normal hexane ( n -hexane), ethyl acetate (EtOAc), normal butanol ( n -BuOH) as shown in FIG. To obtain normal hexane, ethyl acetate, normal butanol, and water fractions, respectively.

4) Standardization by HPLC Analysis of the Rhodiola sagitta Butanol Layer

Experimental Method

5.5 kg of dried ephedra root was ground, extracted three times with ethanol at room temperature, filtered and concentrated. The dried ethanol extract was subjected to solvent fractionation with hexane, ethyl acetate, butanol, and water to obtain 105 g of butanol layer fraction, which was then prepared at a concentration of 250 / ml, followed by 10 injections into the column for HPLC analysis. Was carried out.

Standard Rasveratrol-8- O - β- D-glucoside, torachrysone-8- O - β- D-glucoside, Emodin-8- O - β- D-glucoside and Emodin were dissolved in 50% methanol, Standard solutions for calibration curves were prepared at concentrations of 20, 10, 5, 2.5, and 1.25 / ml, respectively, 10 were injected into a column, and HPLC was performed to obtain chromatograms, and the peak data were plotted to prepare calibration curves.

The HPLC system used was Agilent 1200 HPLC, DAD detector and Chemstation software, and Luna 5C-18 (4.0 × 250 mm) analytical columns from Phenomenex. Sample injection was carried out by using a sample autoinjector, the mobile phase was 1.0 ml / min flow rate using a mixed solution of water and methanol, the wavelength was 290 nm. Gradient began with water: methanol (70:30), and after 35 minutes water: methanol (0: 100), eluted with 100% methanol for 10 minutes. All analytical solvents were HPLC grade solvents (Fisher scientific).

[Experiment result]

As shown in FIG. 2, Rasveratrol-8- O - β- D-glucoside was 10.6 components, torachrysone-8- O - β- D-glucoside 19.46 components, and Emodin-8- O , using HPLC analysis. β- D-glucoside 23.09, Emodin 33.65

In addition, as shown in Table 1, Rasveratrol-8- O - β- D-glucoside was 41.48%, torachrysone-8- O - β- D-glucoside was 2.92%, and Emodin-8- O - β- D-glucoside 36.76 %, Emodin 2.56% was confirmed.

Ingredient Name content Μg / ml Mg / g One Rasveratrol-8- O - β- D-glucoside 10.37 41.48 2 Torachrysone-8- O - β- D-glucoside 0.73 2.92 3 Emodin-8- O - β- D-glucoside 9.19 36.76 4 Emodin 0.64 2.56

One. Ho Jang Geun  Extract and Lineage Separation Powder  Produce

6.8 kg of dry dried jangjanggeun was put in 36 L of ethanol, extracted three times at room temperature for 24 hours, and concentrated under reduced pressure at 40 ° C. to obtain 580 g of ethanol extract.

As shown in FIG. 1, the extract of Hojang-geun extracted by the above method was systematically separated into a layer of normal hexane (46 g), ethyl acetate (250 g), normal butanol (150 g), and water (134 g), and concentrated under reduced pressure. Lyophilization gave a fraction of each layer.

II . Pancreatic lipase ( pancreatic lipase Activity inhibition experiment

Inhibition of porcine pancreatic lipase activity was carried out in vitro using each of the prepared extracts of the extract of jang, hexane, ethyl acetate, butanol and water.

Experimental Method

Pancreatic lipase inhibition experiments were measured using porcine pancreatic lipase. First, 30 μL (10 units) of porcine pancreatic lipase (Sigma, St. Louis, MO, USA) mixed with enzyme buffer [10 mM MOPS (morpholinepropanesulphonic acid) and 1 mM EDTA, pH 6.8] and Tris buffer (100 mM TrisHC1 and 5 mM CaCl ₂ , pH 7.0) 850 μL is prepared by mixing. Orlistat was mixed with 100 μL using Xenical (Roche, Basel, Switzerland) and tested for 15 minutes at 37 ° C. Subsequently, 20 μL of substrate solution [10 mM p- NPB ( p- nitrophenylbutyrate) in dimethylformamide] is added, and then further incubated for 15 minutes. Lipase activity by p -NPB the use of p -nitrophenol the ELISA reader (BIO-TEK, Synergy HT, USA) coming hydrolyzed by the lipase was measured at a wavelength of 400nm. The degree of inhibition of lipase activity was calculated by converting the percentage of absorbance inhibited by the test substance into a percentage value, and the concentration (IC ₅₀ ) that inhibits lipase activity by 50% was calculated.

[Experiment result]

Experiments were carried out according to the experimental example to determine the efficacy of inhibiting pancreatic lipase activity in vitro, and the results are shown in Table 1.

compound Inhibitory effect (IC ₅₀ value) (㎍ / mL) Knotweed Extract 98.97 Hexane layer of extract > 100 Ethyl acetate layer of extract 49.09 Butanol Layer of Rhodiola vulgaris Extract 25.42 Water layer of extract > 100 Xenical 0.16

As shown in the results of Table 1, it was found to have an inhibitory effect on pancreatic lipase, an important enzyme involved in the intestinal absorption of fats from the extracts of K. koji and its various fractions. The 50% inhibition concentration of lipase activity (IC ₅₀ ) was calculated as 25.42 μg / ml in the butanol layer fractions of the root extract, and the inhibitory effect was observed in the ethyl acetate layer and the extract of the root extract. . For the reference drug Xenical, 0.16 μg / ml.

III . Fat emulsion  ( Lipid emulsion ) Short-term Absorption Capacity Analysis

The butanol layer fraction (POCU-1b) of the E. coli muscle extract, which showed the best efficacy in the pancreatic lipase test, was used to measure the ability to inhibit the degradation and absorption of fat in the intestine of rats.

Experimental Method

Four-week-old male wistar rats (Orient Bio, Korea) were prepared for 1 week of purification. After fasting for 18 hours before the experiment, 3 ml Lipid emulsion (3 ml Corn oil, 50 mg cholic acid, 3 ml saline and 1 g cholestryl oleate) and ephedra root butanol layer fraction (POCU-1b) were obtained at final concentrations of 10, 25 and 50 mg / kg. The mixture is administered orally as much as possible, the negative control is administered only the same amount of saline, and the positive control is administered only the lipid emulsion with no drug mixture. As a control drug, Orlistat was administered at a concentration of 200 mg / kg using Xenical (Roche, Basel, Switzerland). Blood is drawn from the cauline for a total of 5 hours immediately prior to drug administration and at 1 hour intervals after drug administration. The collected blood is centrifuged at 5500g for 5 minutes and plasma is separated. Plasma triacylglyceride concentration is measured using the Wako Triglyceride E-Test kit.

[Experiment result]

Experimental results according to the above experimental method are shown in FIG. 3. As a result of experiments to measure the ability of inhibiting short-term fat absorption by mixing and administering a lipid emulsion and a drug, a positive control (positive control) administered only a lipid emulsion, as shown in FIG. Nearly doubled, triglycerides did not increase in the negative control group administered only saline. In addition, the increase in triglycerides was significantly inhibited in the group administered with E. coli butanol fraction (POCU-1b) and Xenical. In addition, all fractions of 10, 25, and 50 mg / kg of KHF were significantly more effective in inhibiting fat absorption than the positive control group, which was administered only lipid emulsion (p <0.01). , 50mg / kg showed the best fat absorption inhibition in the group administered.

IV . In vivo Anti-obesity  efficacy

1) Validation of obesity prevention efficacy in obese animal models In vivo  Fat production inhibition test)

In order to investigate the effect of the increase in the weight gain of butanol layer fraction (POCU-1b) from the extract of Pleurotus eryngii extract, the effect of butanol layer fraction of the extract of Pleurotus eryngii extract on obesity induction was evaluated.

Experimental Method

 The test animals were purchased after 3 weeks old female ICR mouse from Orient Bio Co., Ltd. and used for 1 week. AIN-76A (Research Diets, Inc., D12451, calorie composition: 45% fat, carbohydrate 35%, protein 20%) was used as a high-fat diet for inducing obesity, and the normal diet was Purina Rodent Chow (Purina Korea Inc.). , Product No. 38057, calorie composition: fat 12%, carbohydrate 63%, protein 25%). Feed and drinking water were fed freely. Butanol layer fraction (POCU-1b) of the test drug, root extract, and Xenical, the control drug, were mixed in AIN-76A feed to the following concentrations (0.1%, 0.5%, 1%; Xenical 0.1%). Supplied. The test group was (1) normal diet group (NOR), (2) high-fat diet group (HFD), (3) Xenical administration group, (4) butanol layer fraction (POCU-1b) 0.1% administered group (5) Butanol layer fraction (POCU-1b) 0.5% administration group of the root extract, (6) Butanol layer fraction (POCU-1b) 1% administration group of the root extract. In each group, obesity was induced by feeding a high fat diet mixed with drugs for 10 weeks.

[Experiment result]

The effect of Lipid emulsion and Butanol layer fraction (POCU-1b) of E. coli muscle extract on the induction of obesity in normal rats fed high fat diet was shown. Body weight gain was 3 weeks in the high fat diet group as shown in FIG. The weight gain was observed and the weight gain was observed compared to the group fed the normal diet, and the weight gain of the butanol fraction (POCU-1b) and Xenical feeding groups of the root extract were increased after 4 weeks compared to the high-fat diet group. Has slowed down. After 10 weeks, the changes in body weight at the end of the test showed that the butanol layer fractions of the root extracts of the 0.1, 0.5, and 1% feeding groups had weights of 37.5g, 34.95g, and 34.0g, respectively, compared to 40.1g of the high-fat diet group. In case of Xenical, it was 33.4g, which is similar to that of 1% of Kwon Keun-Geun.

In addition, the visual result of the abdominal fat reduction effect is shown in Figure 5, the result of measuring the fat weight of each part is shown in Figures 6 to 8. Peripheral fat was about 1.1g in the high-fat diet group and increased about 9 times compared to 0.13g in the normal diet group, while 0.9g, 0.6g and 0.5g in the 0.1, 0.5, and 1% feed group of the root extract were The increase was suppressed. Butanol layer fraction of S. aureus extract showed a significant increase inhibition in the 0.5, 1% fed group compared to the high fat diet group (p <0.05). The same result was found for the periphery fat and subcutaneous fat.

2) Obesity treatment efficacy test in obese animal model In vivo  Lipolysis experiment)

To investigate the effect of butanol layer fraction (POCU-1b) from the extract of Pleurotus eryngii (POCU-1b) on the weight loss, the dietary extract of butanol layer (POCU) of the extract of Pleurotus erythematosus in diet-induced obesty (DIO) rats -1b) was administered to observe the change in body weight.

Experimental Method

(1) The test animals were purchased after 3 weeks of age male SD rats were obtained from Orient Bio Co., Ltd. and used for 1 week. AIN-76A (Research Diets, Inc., D12451, Calorie Composition: 45% Fat, Carbohydrate 35%, Protein 20%) was fed high fat diet for 13 weeks to make DIO rats prior to administration of the test drug. As a normal control, Purina Rodent Chow (Purina Korea Inc., Product No. 38057, Calorie Composition: Fat 12%, Carbohydrate 63%, Protein 25%) was used in SD rats of the same age. Feed and drinking water were fed freely. After 13 weeks, DIO rats and normal rats induced high obesity were: (1) normal diet group (NOR), (2) high-fat diet group (HFD), (3) Xenical administration group, and (4) butanol layer The fraction (POCU-1b) 0.1% administered group, (5) Butanol layer fraction (POCU-1b) 1% administration group of the root muscle extract, the test drug POCU-1b and the control drug Xenical at the following concentration ( POCU-1b 0.1%, 1%; Xenical 0.1%) was fed mixed with AIN-76A feed. Each group was fed a high-fat diet mixed with drugs for 7 weeks and observed changes in body weight.

(2) Body weight and dietary intake were measured once a week just before starting test substance administration and during the test period.

(3) After the end of the test, the animals were fasted for 16 hours, and blood was collected from the abdominal aorta at the time of necropsy and centrifuged at 3,000 rpm for 10 minutes to separate serum and plasma. Using the serum, triglyceride, HDL (high density lipoprotein), LDL (low density lipoprotein), and total cholesterol, which are indicators of fat metabolism, were analyzed using a serum autoanalyzer (Hitachi 7060, Japan). Measured.

(4) To examine the weight of the adipose tissue collected at the time of necropsy, take the blood and watch out for the genital fat (left / right), periphery fat (left / right), subcutaneous fat (left / right), and brown fat. The extract was washed with physiological saline, and then water was removed with a filter paper to measure the weight of the adipose tissue.

(5) At the time of necropsy, the extracted adipose tissue was fixed in 10% formalin, embedded in formal, and slide tissues were prepared. H & E staining was performed to analyze the size of fat cells under an optical microscope.

[Experiment result]

1) Body lipolysis efficacy result

To investigate the effect of butanol fraction (POCU-1b) from E. coli muscle extract on weight loss and lipolysis in obese rats induced by high fat diet, high fat diet After induction, the drug was administered to observe the change in body weight. As shown in Figure 9 in the 1% feeding group of butanol layer fractions extract of the roots of the roots of the extract of the high fat diet compared to the feeding group showed a sharp weight loss immediately after administration of the drug (p <0.01), Xenical feeding group In addition, weight loss was shown (p <0.01). However, the weight loss of 1% fed butanol layer fraction of P. aeruginosa extract showed a significant weight loss effect compared to Xenical fed group (p <0.01). There was no weight loss effect.

In addition, as shown in Figures 10 to 13, as a result of measuring the fat weight of each part, the periphery fat was about 11g in the high-fat diet group increased about 2 times compared to 6.2g in the normal diet group, while the butanol layer fractions of the extract In the 0.1% and 1% fed groups, there was no decrease in fat in 11.1g and 6.3g, and the 1% fed group significantly decreased fat (p <0.01). Xenical feeding group also decreased fat weight (p <0.01), but the most significant decrease in fat weight was found in 1% feeding group of butanol layer fraction of S. aureus extract. The same result was found in the periphery fat and subcutaneous fat, and brown fat was also significantly decreased in the 0.1% feed group of the butanol layer fraction of S. aureus extract. (p <0.01)

2) Blood lipid content analysis

The effect of the feed of butanol layer fraction (POCU-1b) of E. coli muscle extract on blood lipid concentration is shown in FIGS. 14 to 17. The total cholesterol concentration increased more than 60% in the high-fat diet group, and in the butanol-fraction fraction fed group of S. aureus, the total cholesterol concentration in blood was lowered in a concentration-dependent manner. This tendency is the same for triglycerides (TG) and LDL cholesterol. Xenical, however, did not reduce triglycerides. HDL cholesterol tended to increase slightly in the feed group and decrease in the butanol layer fraction fed group of the root extract, but not statistically significant.

3) Size analysis of fat cells

As a result of morphological analysis of the adipocytes collected at the time of autopsy, in the group fed high fat diet as shown in Figs. 18 and 19, the size of adipocytes significantly increased more than 2.5 times compared to the normal diet fed group. In this group, the increase in fat cell size was suppressed by 50%. In addition, the increase in fat cell size was also suppressed in the group fed the butanol layer fraction of the root extract, and in the 0.5% feeding group, the fat cell size was higher than the high fat feeding group by 30% and 1% feeding group. The increase of was suppressed (p <0.01).

As described above, the butanol layer fraction of the E. coli muscle extract according to the present invention has excellent anti-obesity effects in inhibiting pancreatic lipase activity, inhibiting intestinal fat absorption, inhibiting weight gain and weight loss, and thus, new obesity and hyperlipidemia and metabolic syndrome. It can be used to treat related diseases such as.

1 is a diagram showing the extraction and phylogenetic separation of the knuckle muscle.

Figure 2 is a graph showing the chromatogram of HPLC of Escherichia coli butanol layer fractions.

Figure 3 is a graph showing the effect of inhibiting the absorption of fat emulsion (lipid emulsion) of the butanol layer fraction of the extract of the roots of the roots.

Figure 4 is a graph showing the effect on the weight gain of the butanol layer fractions of the extract of the roots.

Figure 5 is a visual result photograph of the abdominal fat reduction effect of the butanol layer fractions of the extracts of the roots.

Figure 6 is a graph showing the effect on the genital periphery of the butanol layer fractions of the extract of the root of the root.

Figure 7 is a graph showing the effect on the periphery fat of the butanol layer fractions of the extract of the root of the root.

Figure 8 is a graph showing the effect on the subcutaneous fat of the butanol layer fractions of the extract of the roots.

9 is a graph showing the effect on the weight loss of high-weight obese rats of the extract of Butanol layer fractions

10 is a graph showing the effect on the weight loss of the genital periphery of the high-weight obese rats of the butanol layer fraction of E. coli muscle extract.

FIG. 11 is a graph showing the effect on the peripheral fat weight loss of high obese rats of the butanol layer fraction of Escherichia coli extract.

12 is a graph showing the effect on the subcutaneous fat weight loss of high obesity rats of the butanol layer fractions of the extract of the roots

FIG. 13 is a graph showing the effect of butanol layer fraction (POCU-1b) on the weight loss of brown fat in high obese rats.

FIG. 14 is a graph showing the effect of butanol layer fractions of S. root extract on blood total cholesterol concentration; FIG.

Fig. 15 is a graph showing the effect of butanol layer fractions of S. root extract on blood triglyceride concentration.

Figure 16 is a graph showing the effect of butanol layer fractions of HD extracts on HDL cholesterol concentration.

FIG. 17 is a graph showing the effect of butanol layer fractions of E. coli muscle extract on blood LDL cholesterol concentration; FIG.

FIG. 18 is a graph showing the effect of butanol layer fractions of E. coli muscle extract on morphological changes of adipocytes; FIG.

19 is a graph showing the effect of the butanol layer fractions of the E. coli muscle extract on the change of adipocyte size.

Claims (10)

In the extract of Hojang-geun, which is obtained by extracting Kho-Geun-Geun with an alcohol or an aqueous solution of alcohol, filtering the extracted extract and filtration under reduced pressure, A pharmaceutical composition for preventing or treating obesity, containing the extract of E. coli as an active ingredient. In the extract of Hojang-geun, which is obtained by extracting Kho-Geun-Geun with an alcohol or an aqueous solution of alcohol, filtering the extracted extract and filtration under reduced pressure, Obesity prevention and improvement health functional food containing the extract of the extract as an active ingredient. Ethyl acetate of the extract of e.g. root extract, which is extracted by filtering the extracted extract with an alcohol or an aqueous solution of alcohol, and filtering the extracted extract, and extracting the e.g. extract from normal hexane, ethyl acetate, butanol, and water in order. In the layer fraction, A pharmaceutical composition for the prevention or treatment of obesity, containing the ethyl acetate layer fraction of the extract of E. coli as an active ingredient. Butanol layer fraction of K. root extract, which is extracted by filtering the extracted extract with an alcohol or aqueous alcohol solution, filtering the extracted extract and concentrated under reduced pressure, and then separating the extract from the extract of normal root hexane, ethyl acetate, butanol, and water in the order of butanol. To Obesity prevention and improvement health functional food containing the ethyl acetate layer fraction of the extract of the extract Butanol layer fraction of K. root extract, which is extracted by filtering the extracted extract with an alcohol or aqueous alcohol solution, filtering the extracted extract and concentrated under reduced pressure, and then separating the extract from the extract of normal root hexane, ethyl acetate, butanol, and water in the order of butanol. To A pharmaceutical composition for the prevention or treatment of obesity, comprising butanol layer fraction of the extract Jangeun root as an active ingredient. Butanol layer fraction of K. root extract, which is extracted by filtering the extracted extract with an alcohol or aqueous alcohol solution, filtering the extracted extract and concentrated under reduced pressure, and then separating the extract from the extract of normal root hexane, ethyl acetate, butanol, and water in the order of butanol. To A dietary supplement for the prevention and improvement of obesity, comprising butanol layer fraction of the extract Jangeun-geun as an active ingredient. Butanol layer fraction of K. root extract, which is extracted by filtering the extracted extract with an alcohol or aqueous alcohol solution, filtering the extracted extract and concentrated under reduced pressure, and then separating the extract from the extract of normal root hexane, ethyl acetate, butanol, and water in the order of butanol. To A pharmaceutical composition for preventing or treating hyperlipidemia, comprising the butanol layer fraction of the extract of E. coli as an active ingredient. Butanol layer fraction of K. root extract, which is extracted by filtering the extracted extract with an alcohol or aqueous alcohol solution, filtering the extracted extract and concentrated under reduced pressure, and then separating the extract from the extract of normal root hexane, ethyl acetate, butanol, and water in the order of butanol. To Health functional food for preventing and improving hyperlipidemia containing butanol layer fractions of the extract of the extract Keun-Geun as an active ingredient. Butanol layer fraction of K. root extract, which is extracted by filtering the extracted extract with an alcohol or aqueous alcohol solution, filtering the extracted extract and concentrated under reduced pressure, and then separating the extract from the extract of normal root hexane, ethyl acetate, butanol, and water in the order of butanol. To Pharmaceutical composition for the prevention or treatment of metabolic syndrome containing butanol layer fraction of the extract Kwon Keun-Geun as an active ingredient. Butanol layer fraction of K. root extract, which is extracted by filtering the extracted extract with an alcohol or aqueous alcohol solution, filtering the extracted extract and concentrated under reduced pressure, and then separating the extract from the extract of normal root hexane, ethyl acetate, butanol, and water in the order of butanol. To Health functional foods for preventing and improving metabolic syndrome containing butanol layer fraction of the extract of the jangeungeun as an active ingredient.

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