KR101530135B1 - A composition for anti-obesity comprising an extract of starfish - Google Patents

Abstract

The present invention relates to an anti-obesity composition comprising a starfish extract, which comprises an anti-obesity composition comprising a glycolipid component of starfish extract and a composition for improving obesity comprising a glycolipid component of starfish extract. The starfish extract of the present invention is preferably used as a food or a pharmaceutical composition used for prevention and treatment of obesity because the glycolipid component of the extract shows anti-obesity activity. Further, it can be used as a food or a pharmaceutical composition for obesity, And cerebrovascular disorders. ≪ Desc / Clms Page number 2 >

Description

[0001] The present invention relates to an anti-obesity composition comprising starfish extract,

The present invention relates to an anti-obesity composition comprising a starfish extract, which comprises an anti-obesity composition comprising a glycolipid component of starfish extract and a food composition for improving obesity comprising the composition.

In recent years, with the improvement of the economic level, the overweight and obesity populations are increasing widely due to the increase of calorie intake and lack of exercise, and it is becoming a social problem gradually. This phenomenon is problematic not only for adults but also for all children and adolescents. In particular, the problem is even more serious in that childhood and adolescent obesity is likely to lead to adult obesity.

 The RAND Research Institute of the United States randomly selected about 10,000 men and women from all over the United States to examine the quantitative relationship between health risk factors such as smoking, drinking, poverty and obesity and 17 chronic diseases such as cancer, heart disease, stroke, arthritis, hypertension, , Obesity was found to cause more chronic diseases than drinking, poverty, and smoking in most cases. Thus, prevention and treatment of obesity is a very important issue in modern society because obesity threatens quality or quantity of life without special hereditary diseases.

On the other hand, in Korea, the westernization of dietary habits changed to high fat, high protein, and high calorie diet, and the problem of obesity began to arise due to foreign fast food inflow. Recently, according to the shortage of modern people, obesity and accompanying adult diseases are increasing rapidly. According to the data of the National Statistical Office 2000, deaths due to cardiovascular diseases were ranked as the number one cause of hypertension and hyperlipidemia with the increase of obesity population over the past decade (1990 ~ 1999).

Obesity is caused by a variety of factors, such as eating habits, lack of activity, genetic factors, hormonal factors, central nervous system disorders, psychological disorders, and social, cultural, and economic factors, rather than being caused by any one cause. In addition, such obesity can cause various chronic degenerative diseases such as hypertension, type 2 diabetes, fatty liver, hyperlipidemia, degenerative arthritis, and some cancer diseases. That is, in the case of obesity, blood glucose and blood pressure rise, hypertension and diabetes are likely to occur, heart diseases such as angina pectoris occur well, and fat accumulation in liver and gallbladder leads to the risk of hyperlipemia, arteriosclerosis, fatty liver and gallstone . In addition, degenerative arthritis can be caused by the burden of joints due to heavy weight, and uric acid generated by metabolism of fat may accumulate in the joints and cause joint pain. In addition, women are known to cause menstrual irregularities and infertility, malnutrition in men, and sexual dysfunction. In addition, cancer incidence such as cancer of the breast or breast cancer is reported to be very high. Especially, obesity patients can cause severe mental disorders due to social disorder or inferiority.

Accordingly, there is a need for an effective therapeutic agent for treating or preventing obesity in order to prevent a number of complications caused by the obesity phenomenon itself. However, there are few treatments that can directly treat obesity. Exercise and diet are recommended only. However, the prevention of obesity through this method and the fundamental treatment effect of obesity are very low. In addition, medicines that are marketed as some obesity treatment drugs have side effects caused by platelet aggregation inhibitory action and vein expansion, or serious side effects that cause mental disorders such as depression, which are not recommended. In addition, health functional agents and adjuvants may be used as adjunctive therapies, but they are not a fundamental treatment for obesity. Therefore, it is inevitable to develop a composition having excellent anti-obesity activity while exhibiting excellent effects.

As a result, the inventors of the present invention have found that the glycolipid component of starfish extract not only reduces the concentration of triglyceride in blood but also exhibits anti-cholesterol activity and exhibits anti-obesity activity And the present invention has been completed.

It is an object of the present invention to provide an anti-obesity composition comprising a glycolipid component of a starfish extract.

Another object of the present invention is to provide a food composition for improving obesity comprising a glycolipid component of starfish extract.

In one aspect, the invention relates to an anti-obesity composition comprising a glycolipid component of a starfish extract.

In the present invention, the term "sea star, starfish" is a generic term belonging to echinoderm and starfish, and it is known that most papers have five regenerable arms in a radially symmetrical form. Starfish has a rigid exoskeleton of echinoderms, and behind the exoskeleton can be used to take food and eat digestive juices while holding a game. Starfish species are varieties, or some are hermaphrodites.

Examples of the starfish include Asterias amureasis, Ophioplocus japonicus, Certonardoa semiregularis, Asterina pectinifera, Sun starfish (starfish), starfish starfish Solarstar borealis, Solarstar paxillatus, Astropecten polyacantbus, and Marthasterias glaciallis. However, the types of starfish that can be used as an extract of the present invention are not limited by the above examples.

It is generally known that the constituents of starfish have lower water content and higher ash content than ordinary seafood. For example, the starfish, which is known to be the most abundant, has a moisture content of about 67% to 68% and a solid content of about 32% to 33%, and ashes account for more than half of the solids. Organic matter of starfish is known to occupy about 13 ~ 14%, and organic matter is composed of about 10% of crude protein, about 3.7% of crude fat, and about 0.2% of carbohydrate.

On the other hand, the starch contains inorganic substances such as calcium (Ca), phosphorus (P), potassium (K) and magnesium (Mg), and about 30.2%, 1.4% 1.6% and 2.1%, respectively.

ingredient Moisture content Solids Organic matter Ash starfish 67.8 32.2 13.9 18.3

These starfish have been developed as feed and fertilizer additives for animals and plants due to the above-mentioned constitutional characteristics, and are sometimes used as calcium supplement in humans. In addition, some antioxidative and antimicrobial activities against starfish organic solvent extracts have been reported. In addition, certain components of starfish extracts can be used as fish attractants, and saponin components in starfish extracts are known to inhibit cancer activity and cancer cell proliferation.

However, the fact that starfish extract can directly inhibit or treat obesity through weight loss, reduced cholesterol concentration, and reduced blood lipid is not known. The inventors of the present invention firstly found that glycolipids in starfish extracts can exhibit anti-obesity activity through the above-mentioned action.

'Glycolipid', a component of starfish extract, is a kind of complex fat, meaning a compound in which sugar and lipids are covalently bonded. The O- antigen of Gram-negative bacteria is divided into two types, one consisting of complex polysaccharides and the one consisting of oligosaccharides. Usually, the latter refers to the latter. It is widely distributed in plants, animals and microorganisms, and has a chemical structure including lipids which are commonly sphingosine and glycerol. Unlike phospholipids, glycolipids contain saccharides or nitrogen compounds that are not phosphoric acid. These glycolipids constitute the constituents of the cell membrane in the human body, and they act as signal transducers by recognizing signals outside the cells.

For purposes of the present invention, the glycolipid component of the present invention means a glycolipid component isolated from the starfish extract, preferably a glycolipid component separated from the starfish ethyl acetate fraction, more preferably three hexadecane and one May be a glycolipid component comprising ceramide.

In the present invention, it was confirmed that the glycolipid component contained in the starchy ethyl acetate extract fraction reduced triglyceride accumulated in the adipose tissue and decreased the LDL-cholesterol concentration to have anti-obesity activity. The MMP- 9 (FIGS. 4 and 5 and Table 4).

The term " extraction " of the present invention is a method of separating components using a solvent capable of dissolving a specific substance in a mixture of two or more substances, using a chemical reaction such as an acid- Only the difference in solubility may be used. Solid-liquid extraction is referred to as solid-liquid extraction, liquid extraction is referred to as liquid-liquid extraction, and solid-liquid extraction as leaching.

In the extraction method used in the present invention, a solid-liquid extraction method using a difference in solubility of a sample material in an organic solvent is used. Preferably, only a soluble fraction is obtained by adding a nucleic acid to a fraction extracted from a methanol component of starfish, After extracting only the components dissolved in ethyl acetate after treatment with ethyl acetate to obtain a component exhibiting an effective effect.

The term " obesity " of the present invention means a state in which the amount of fat tissue in the body is excessively increased. Such obesity is caused by an imbalance in the synthesis and degradation of fat in the adipocyte. Obesity resulting from the accumulation of fat more than necessary increases the risk of developing a chronic degenerative disease. In particular, abdominal obesity is caused by coronary artery disease, hypertension, stroke, The risk of

 Obesity is divided into simple obesity and secondary obesity. Simple obesity accounts for about 95% of obese people because of obesity due to overeating and lack of exercise, without special hereditary diseases, endocrine diseases, neurological abnormalities. Secondary obesity is obesity as a result of some diseases such as endocrine obesity, hereditary obesity, hypothalamic obesity, frontal obesity, and metabolic obesity. Obesity can be treated by treating the disease.

In addition, obesity can be classified into fat cell proliferative obesity, fat cell non-large obesity, and mixed fat type obesity according to the size and number of adipocytes. Adipocyte proliferative obesity is caused by an increase in fat cell count, And usually occurs from childhood and early childhood through maturity. Obesity is a cause of overweight and energy imbalance in the degree of obesity in childhood obesity, and childhood obesity is shifted to adult obesity, the number of fat cells is three to four times the number of people with normal weight, It is known that treatment is difficult because it is difficult to reduce the number of cells themselves.

In adipocyte non-large-sized obesity, the number of adipocytes is almost normal, but each adipocyte is accompanied by hypertrophy of cells growing in size. Usually non-large-sized obesity starts after becoming an adult, and it becomes fattening after middle age, and it is common in pregnancy. However, non-large-sized obesity can be regarded as easier than proliferative obesity because the size of the fat cells is reduced when the body weight is reduced and the fat cells are returned to the original state. Adipocyte mixed type obesity is an obesity that increases in both the fat and body size and the size.

The composition of the present invention is able to inhibit obesity in both cases of adipocyte proliferation type obesity and adipocyte non-large type obesity, and is effective in reducing visceral fat, which is a major complication (Fig. 3).

 The term 'cholesterol' is widely used as a cellular component of higher animals and is a steroid compound. It is a major component of the membrane system of the cell together with the phospholipids, and plays a major role in membrane structure and function. It is a form of animal fat that is essential for our body. It is a nutrient which is a component of cell membrane, hormone, and digestive juice. Cholesterol is divided into high-density lipoprotein (HDL) and low-density lipoprotein (LDL), which are known to cause diseases such as hyperlipemia and atherosclerosis when the level of harmful cholesterol such as LDL-cholesterol is increased.

Low-density lipoprotein (LDL) cholesterol plays a role in transporting cholesterol produced by the liver to the whole body's cells. However, accumulating more LDL-cholesterol in the cells of the arterial wall may cause arteriosclerosis. Accumulation of LDL in the body by more than 50% causes arteriosclerosis and thus a heart attack. High density lipoprotein (HDL), on the other hand, is responsible for removing cholesterol from the arterial wall cells and returning it to the liver. HDL-cholesterol is a small particle and cleans up the blood to remove the fat component, so a lot of HDL-cholesterol arteriosclerosis is not good because it is called good cholesterol.

The extract of the present invention reduces blood LDL in the body and can control the concentration of cholesterol which is a cause of obesity by comparing or increasing HDL (Tables 3, 4 and 4).

On the other hand, fatty acids include saturated fatty acids and unsaturated fatty acids. Saturated fatty acids do not have double bonds between carbon atoms, while unsaturated fatty acids have one or more double bonds. When the concentration of saturated fatty acids in the blood increases due to obesity, it converts saturated fatty acids into cholesterol in the liver, resulting in an increase in blood LDL-cholesterol, which causes atherosclerosis, heart attack, stroke and dementia.

The term " visceral fat " of the present invention means fat existing between organs and is distinguished from subcutaneous fat accumulated below the skin layer. Particularly, the most related site of metabolic abnormalities in body fat distribution is the fat accumulating in the internal organs of the upper abdomen. This abdominal fat is more rapidly degraded than the fat cells of the other regions, and the amount of free fatty acid secreted increases It is known to cause metabolic disorders. In the case of visceral fat, it accumulates more easily than subcutaneous fat and easily flows into the blood, thereby increasing the blood cholesterol level, generating harmful free radicals, and visceral fat sticking around the organ, do. In addition, the fat permeated into the blood lowers the insulin effect in the body, disturbing the influx and action of glucose, causing diseases such as hyperinsulinemia, hypertension, diabetes, and dyslipidemia, and a combination of different mechanisms, , Angina pectoris, and myocardial infarction.

It was confirmed that the extract of the present invention can induce a substantial anti-obesity effect by effectively inhibiting visceral fat, which is a major cause of not only obesity but also major diseases (FIG. 3).

The term " triglycerides " of the present invention is one of the lipid components in the blood, and exists together with cholesterol, triglyceride, phospholipid, free fatty acid and the like in the blood. Since triglyceride in blood is free from blood, it can not be transported by itself. Therefore, in order to move to a necessary place in the body, it is dissolved in blood and binds to apo-protein to form lipoprotein. In most cases, triglyceride produces LDL-cholesterol and HDL - It is known to promote cholesterol degradation.

Preferably, the starfish extract of the present invention can prevent or treat obesity and thus serious complications by preventing the generation of LDL-cholesterol that starts from obesity and causes serious diseases such as heart attack or arteriosclerosis as described above .

In addition, the composition of the present invention can achieve an anti-obesity effect by inhibiting MMP-9 (matrix metalloproteinase-9) of mast cells.

The term "matrix metalloproteinase (MMP)" of the present invention refers to an extracellular matrix metalloproteinase, which is a cell composed of structural proteins such as collagens, proteoglycans and glycoproteins It is known to play an important role in the degradation process of the extracellular matrix (ECM). These MMPs are numbered according to their types and up to 20 or more MMPs (MMP-1, MMP-2, MMP-3, MMP-4, MMP-5, MMP-6, MMP- MMP-11, MMP-12, MMP-13, MMP-14, MMP-15, MMP-16, MMP-17, MMP-18, MMP- , MMP-21, MMP-22 and MMP-23).

In addition to histamine, tryptase, chymase, heparin, interleukin, tumor necrosis factor α (TNF-α), transforming growth factor (TGF-β), basic matrix growth factor (bFGF) , Which are known to produce vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP-9). MMP-9 increases the amount of MMP-9 as mast cells increase, so that it infiltrates surrounding tissues and penetrates blood vessels and promotes diseases such as arteriosclerosis. Accordingly, it is known that when the mast cell is increased, the MMP-9 is also increased proportionally. In the present invention, the extract of the present invention, preferably the glycolipid component of the extract of the present invention inhibits MMP-9 activity of mast cells Confirming that it has anti-obesity activity (FIG. 5).

In the present invention, in order to confirm the anti-obesity activity of the starfish ethyl acetate fraction, the experimental mice were divided into 5 groups, and the anti-obesity activity was observed by administering the starfish ethyl acetate fraction of the present invention after the general diet and the high fat diet, respectively. As a result, it was confirmed that weight loss, reduction of LDL-cholesterol level in blood, decrease of total-cholesterol level, and inhibition of MMP-9 activity were observed in a high fat diet mouse (Table 4 and Figs.

In order to confirm which of the fractions has substantial activity, the present inventors used the ethyl acetate fraction in a solvent system of chloroform: methanol: water = 30: 60: 8 using high performance thin layer chromatography (HPTLC) As a result of the re-fractionation, four glycolipid components (T1, T2, T3 and T4) were separated according to the mobility. Among these components, the portions having MMP-9 activity were T1 and T2 (Fig. 5 to Fig. 8).

Further, the ethyl acetate fraction was dissolved in chloroform and fractionated with chloroform: methanol = 8: 2 using TLC. As a result, a glycolipid component of a single band product was obtained (FIG. 9) : Water = 60: 35: 8 to obtain a single glycolipid component. The result of MALDI-QIT-MS analysis showed that the glycolipid had three hexoses and one ceramide structure (FIG. 10).

In another aspect, the present invention relates to a food composition for improving obesity comprising glycolipid of starfish extract as an active ingredient.

The term " improvement " of the present invention means an effect of preventing or alleviating symptoms associated with obesity by administering a health functional food composition containing a starfish extract.

The food composition of the present invention includes all forms of functional food, nutritional supplement, health food, and food additives, and the food composition of this type is known in the art And can be prepared in various forms according to known conventional methods. For example, as a health food, the starfish extract of the present invention may be prepared in the form of tea, juice, and drink and then taken for drinking, granulated, encapsulated and powdered. However, It is not.

In addition, the health functional food of the present invention may contain additional additives, and the kind thereof is not particularly limited. Preferable examples of the functional food that can be added include drinks (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruits, bottled, jam and marmalade), fish, meat and processed foods (Such as butter and cheese), edible vegetable oil, margarine, vegetable protein (such as corn oil), breads and noodles (such as udon, buckwheat noodles, ramen noodles, spaghetti and macaroni), juice, various drinks, cookies, , Retort food, frozen food, various kinds of seasoning (for example, miso, soy sauce, sauce, etc.) by adding the starfish extract of the present invention.

Preferably, the food composition for improving obesity of the present invention may be an ethyl acetate fraction of a starfish extract, more preferably it may be a glycolipid component of the ethyl acetate fraction, and even more preferably contains three hexoses and one ceramide Lt; / RTI > The present inventors confirmed that the starfish ethyl acetate fraction was administered to obesity mice induced by high fat diet and showed anti-obesity activity such as weight loss and visceral fat loss, and it was confirmed that the above-mentioned activity was due to the glycolipid component .

The inventors of the present invention divided the four active components (T1, T2, T3, and T4) according to the mobility to separate active ingredients of the extract of the present invention. T1 and T2 fractions, Preferably, the component of the T2 fraction can induce anti-obesity activity through inhibition of MMP-9 activity. It was also confirmed that the glycolipid component contained three hexoses and one ceramide.

The starfish extract of the present invention is preferably used as a food or a pharmaceutical composition used for prevention and treatment of obesity because the glycolipid component of the extract shows anti-obesity activity. Further, it can be used as a food or a pharmaceutical composition for obesity, And cerebrovascular disorders. ≪ Desc / Clms Page number 2 >

FIG. 1 is a schematic view of a fractionation process of a starfish extract. FIG.
FIG. 2 is a graph showing the weight loss effect of raw mouse fed with high fat diet (HD) by starfish extract ingestion.
FIG. 3 is a graph showing the weight reduction effect of mouse abdominal fat tissue according to the concentration of the starfish extract of the present invention (starfish extract: SE, ND: normal diet, HD: high-fat diet) .
FIG. 4 is a graph showing changes in blood total-cholesterol and LDL-cholesterol according to the administration of the starfish extract of the present invention in a high cholesterol diet animal model. (A) total-cholesterol (B) LDL-cholesterol, ND: normal diet, HD: high fat diet.
FIG. 5 is a graph showing the inhibitory activity of MMP-9 production of T1 and T2 isolated by fractionation and sub-fractionation by HPTLC of ethyl acetate (EtOAc) fraction. TNF-a-treated arterial vascular smooth muscle cells (HASMC).
6 to 8 are diagrams showing the determination results of the structures of T1, T2, T3 and T4 compounds by Maldi-Tof analysis. The T2 fraction was considered to be a single peak due to a single peak, and it was confirmed through experiments that T2 had MMP-9 inhibitory activity at the same time.
FIG. 9 is a graph showing the glycolipid component of a single band product through TLC analysis after dissolving the ethyl acetate (EtOAc) fraction in chloroform.
10 is a graph showing the results of MALDI-QIT-MS analysis of single glycolipids separated through HPLC. Indicating that the glycolipid component contains three hexoses and one ceramide (6-per-6-per-6-per-ceramide).

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

Example  1: Starfish fraction extraction

In the present invention, in order to confirm the pharmacological action of starfish on hyperlipidemia and obesity, an extraction process for obtaining an active ingredient of starfish is shown by a schematic diagram (Fig. 1). Specifically, 10 kg of starfish (National Bioscience Institute of Bioscience and Biotechnology) extracted from the southern coast of Korea was extracted with methanol to obtain 1.4 kg yield. Of these, 40 g of the methanol extract was fractionated into water, hexane, ethylacetate (EtOAc) and butanol (BuOH). As a result, 5.301 g of dried nucleic acid extract, 0.308 g of dried ethyl acetate extract, 2.465 g of dried butanol extract and 19.6 g of water-soluble fraction were obtained. The extracts for each solvent were lyophilized and dissolved in DMSO at a concentration of 100 mg / ml.

Example  2: ingestion of starfish extract, weight and biochemical changes in experimental animals

To investigate the effect of starfish extract on weight gain of mice due to high calorie diet, 4 - week - old C57BL / 6J mice were divided into 5 groups. The animals used in the experiments were managed according to the animal study guidelines. Experiments were carried out on 4-week-old C57BL / 6J mice between 17 and 20 g body weight in five groups of control, high fat diets, and 1 mg / kg, 10 mg / kg and 100 mg / . The light / dark cycle of the animal control room was 12h / 12h and the temperature was maintained at 22 ° C. The high-fat diet (HD) used in the positive control group was D12492 (60 kcal fat, 20 kcal protein, 20 kcal carbohydrate: casein 200 g, L-cysteine 3 g, maltodextrin 125 g, sucrose 68.8 g, 50 g of cellulose, 25 g of soybean oil, 245 g of lard, and other 57.05 g) were used.

Oral doses of starfish extracts for experimental mice were given daily at doses of 1 mg / kg, 10 mg / kg, and 100 mg / kg. The regular diet and high fat diets used were consistently supplied as 5g / mouse ㆍ day ^{- 1} . The body weight of the experimental mice was measured every 5 days. Fifty days after the experiment, the experimental mice were sacrificed and the serum was separated and sacrificed. The epithelial adipose tissue and the peritoneal and retroperitoneal adipose tissues were cut and weighed.

According to the method performed in Example 2, after 50 days, the body weights were 27g and 34g on average, and a weight difference of about 7g was obtained as shown in Table 2. In addition, mice receiving 1 mg / kg of starfish extract showed an average body weight of about 30 g, 50 mg / kg, about 25 g, and 100 mg / kg after 50 days, mg / kg of starfish extract showed weight loss effect, and maximum activity was observed within the range of 1 mg / kg to 10 mg / kg (Table 2).

When the starfish extract was administered over 50 days, changes in body weight of the experimental mice were shown in FIG. 2 as time elapsed. Compared to the control group, the body weight was decreased in the 10 mg / kg and 100 mg / kg groups (FIG. 2).

Example  3: Blood lipid concentration and Leptin ( leptin Effect of Starfish Extract on Concentration

To investigate the effect of starfish extract on blood lipid concentration, the contents of total-cholesterol, HDL-cholesterol, LDL-cholesterol and triglyceride were investigated by using serum from a laboratory mouse administered with a starfish extract for 50 days.

The concentration of leptin in the serum was determined using a mouse leptin ELISA kit from SHBAYAGI Co. (Shibayagi Co. Ltd. 1062-1, Ishihara, Shibukawa, JA). Measurement of total-cholesterol, HDL-cholesterol, and triglyceride in the serum was performed using asan set of Asan pharm. Co. Ltd., Hwaseong, Gyeonggi-do, Korea. LDL-cholesterol in the serum was calculated by the formula of {(total-cholesterol) - (HDL-cholesterol) - (Triglyceride / 5)}.

As shown in Table 3, total cholesterol, HDL-cholesterol, LDL-cholesterol and triglyceride contents were higher in the high fat diet group than in the control group (ND) (HD SE1), and 10 mg / kg or 100 mg / kg was slightly lower than that of high-fat diets (HD SE0). Similarly, total-cholesterol content was significantly reduced at 1 mg / kg and slightly decreased at 10 mg / kg and 100 mg / kg (Table 3).

HDL-cholesterol was not significantly different between the HD group (HD SE0) and the starfish extract-treated group, and the triglyceride group did not show any significant difference compared with the HD group and slightly increased in the 100 mg / kg group Table 3).

Leptin is secreted from adipose tissue and is known to be associated with obesity (Skurk et al.). In the present study, leptin was increased about 4 times in the high fat diet compared to the control group (ND), but decreased to 1.5 to 2 times in the experimental mouse group treated with starfish extract (Table 3).

Example  4: Effect of starfish extract on abdominal subcutaneous fat and visceral fat

Starfish extract was administered to experimental mice for 50 days and the weight of abdominal subcutaneous fat and visceral fat of normal diet (ND) and high fat diet group (HD) were measured and compared.

As a result, as shown in FIG. 3, the abdominal subcutaneous fat of the high fat dietary group was increased to about 1.7 g, but in the starfish extract-treated group, it was dose-dependent decreased to about 0.5 g. And increased to about 2.5 g in the diet group, but decreased to 0.5 g in the starfish extract-treated group (Fig. 3).

Example  5: starfish ethyl acetate ( EtOAc ) Fractional blood cholesterol and triglyceride changes after mouse diet

In order to observe changes in mouse blood lipid levels by starch ethyl acetate fraction, 4 - week old mice were divided into 6 groups of dietary, high fat diets, and high fat diets + EtOAc groups for 6 weeks, and blood cholesterol and triglycerides Were observed. These measurements were carried out by ELISA using reagents from Asan Pharmaceutical Co., Ltd. and the like.

As a result, the total-cholesterol concentration in the high fat dietary group + EtOAc fraction group was lower than that in the high fat dietary group, and the HDL-cholesterol was slightly increased in the high fat dietary group + EtOAc fraction intake group (Fig. 4 and Table 4).

In addition, LDL-cholesterol levels were also lower in the high-fat diet + EtOAc fraction group than in the high fat diet group. The concentration of triglyceride was not significantly different from that of the high fat diet group (FIG. 4 and Table 4).

These results suggest that ethyl acetate fraction of starfish extract can inhibit not only obesity but also vascular diseases.

Example  6: TLC ( thin layer chromatography ) Starfish using the technique EtOAc  As the active ingredient in the fraction Glycolipid  Identification of ingredients

The EtOAc fraction was fractionated using HPTLC (high performance thin layer chromatography). A solvent system of chloroform: methanol = water = 30: 60: 8 was used. EtOAc fraction was dissolved in the same solvent, and the mixture was divided into 4 compounds according to mobility. As a result, After dissolving in the same solvent, they were named as sT1, sT2, sT3, sT4.

Dried using an evaporator, and dissolved in DMSO to determine activity. As shown in FIG. 5B, when the fraction of T1-T4 was treated with the fraction of arterial smooth muscle cells and activated by TNF-α, the activity was examined by zymography after 24 hours. As a result, MMP-9 inhibitory activity (Fig. 5).

The mobility of the four mixtures fractionated according to the method performed in Example 6 was compared with that of the ganglioside standard using Maldi-Tof, and the T2 of the glycolipids Tl and T2, in which anti-smooth muscle cell activity was confirmed, was purely isolated 6 to 8).

Example  7: Acquired from starfish ethyl acetate fraction Glycolipid  Identification of structure of ingredients

To determine the structure of glycolipids in starfish extract, the starfish ethyl acetate fraction was dried and dissolved in chloroform. Lipids and glycolipids were then separated by a silica gel column. To this end, the supernatant was adsorbed onto 1 g of a Waters silica gel disposable column and washed with chloroform (C). The fractions were then fractionated at a ratio of C: M (chloroform: methanol) 9: 1, 8: 2, 7: 3, 1: 1 and dried up with nitrogen to confirm by TLC. As a result, a glycolipid component of a single band product was obtained at 8: 2 (FIG. 9).

For the HPLC analysis of the fractions obtained at the ratio of C / M 8: 2, the glycolipids were additionally eluted from the column AQUASIL ss-1251 4.6 Ø × 250 mm with a solvent of chloroform: methanol: water = 60: 35: 8. MALDI-QIT-MS (Shimazu Co.) analysis of the thus obtained single glycolipids revealed glycolipids having three hexoses and one hexamide-hexose-ceramide structure (FIG. 10).

Claims (4)

(a) repeatedly extracting the starfish three times with methanol for 24 hours to obtain a methanol extract;
(b) adding water and hexane to the methanol extract obtained in the step (a) to separate into a hexane layer and a water layer;
(c) fractionating the aqueous layer obtained in the step (b) with ethyl acetate, separating into an ethyl acetate layer and an aqueous layer, and obtaining an ethyl acetate layer therefrom to obtain an ethyl acetate fraction;
(d) drying the ethyl acetate fraction obtained in the step (c), dissolving it in chloroform, and adsorbing it on a silica gel column; And
(e) washing the column with chloroform, and then adding a mixed solvent of chloroform-methanol having a ratio of chloroform: methanol of 8: 2 (v: v) to the column to obtain an eluate A pharmaceutical composition for preventing or treating atherosclerosis, comprising a glycolipid comprising three hexoses and one ceramide extracted from a starfish.
The pharmaceutical composition according to claim 1, wherein the composition has matrix metalloproteinase-9 (MMP-9) inhibitory activity in arterial smooth muscle cells.
(a) repeatedly extracting the starfish three times with methanol for 24 hours to obtain a methanol extract;
(b) adding water and hexane to the methanol extract obtained in the step (a) to separate into a hexane layer and a water layer;
(c) fractionating the aqueous layer obtained in the step (b) with ethyl acetate, separating into an ethyl acetate layer and an aqueous layer, and obtaining an ethyl acetate layer therefrom to obtain an ethyl acetate fraction;
(d) drying the ethyl acetate fraction obtained in the step (c), dissolving it in chloroform, and adsorbing it on a silica gel column; And
(e) washing the column with chloroform, and then adding a mixed solvent of chloroform-methanol having a ratio of chloroform: methanol of 8: 2 (v: v) to the column to obtain an eluate A composition for improving atherosclerosis, comprising a glycolipid comprising three hexoses and one ceramide derived from a starfish.
4. The composition of claim 3, wherein the composition has matrix metalloproteinase-9 (MMP-9) inhibitory activity in arterial smooth muscle cells.

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