KR102239066B1 - Composition for preventing, ameliorating or treating metabolic diseases comprising mixture of plant extract as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating metabolic diseases containing a mixture of plant extracts as an active ingredient, and the mixture of bellflower extract, celery extract and green tea extract of the present invention is a high fat diet animal model in obesity, hyperlipidemia, The composition of the present invention, which contains a mixture of bellflower extract, celery extract, and green tea extract as an active ingredient, is effective for improving arteriosclerosis, cardiovascular disease, hyperinsulinemia, and fatty liver. It can be usefully used as a health functional food composition or as a material for animal feed.

Description

Composition for preventing, ameliorating or treating metabolic diseases comprising mixture of plant extract as effective component}

The present invention relates to a composition for preventing, improving or treating metabolic diseases containing a mixture of plant extracts as an active ingredient.

With recent economic development and changes in dietary habits, the onset of metabolic diseases, including various diseases such as obesity, hyperlipidemia, high blood pressure, arteriosclerosis, hyperinsulinemia, diabetes or liver disease, is rapidly increasing. Each of these diseases may occur, but in general, most of them are closely related to each other and are accompanied by various symptoms.

Obesity is widely known to cause chronic diseases such as fatty liver, high blood pressure, diabetes, and cardiovascular disease. As of 2007, 1.7 billion people, which account for about 25% of the world's population, are overweight (BMI> 25), and children 5 One out of the population falls under pediatric obesity, and the number of pediatric obesity is rapidly increasing, making it a serious social problem. As an obesity treatment agent sold at home and abroad, there is'Zenical', whose main ingredient is orlistat, which has been approved by the U.S. FDA, but Xenical, which inhibits lipase action, has side effects of the gastrointestinal system such as fat stool, gas production, and decreased absorption of fat-soluble vitamins. There is this.

Hyperlipidemia is a condition in which more fatty substances are present in the blood than necessary and accumulates on the walls of blood vessels, causing inflammation, and as a result, cardiovascular disease. As lipid components such as cholesterol in the blood increase, blood flow becomes impaired, and as lipid components adhere to the arterial wall, chronic inflammatory reactions and arteriosclerosis are triggered. It can cause myocardial infarction, stroke or cerebral infarction.

In the arteriosclerosis, fatty substances (plaque) containing cholesterol, phospholipids, calcium, etc. are accumulated in the inner membrane of blood vessels to reduce the elasticity and harden the arteries, and the blood supply is inhibited due to the narrowed arteries or the pressure is increased, causing rupture, detachment, etc. It refers to the state in which it is happening.

Currently, the treatment of hyperlipidemia or arteriosclerosis is a'statin' series of drugs that have inhibitory activity against HMG-CoA reductase, which plays an important role in the synthesis of cholesterol in the liver. It is known to have side effects such as muscle toxicity.

Hyperinsulinemia is a condition associated with obesity or diabetes, such as promoting sympathetic activity or promoting sodium absorption in the kidneys, as a condition with high insulin levels in the blood. There is an urgent need to develop a therapeutic agent that can lower the level.

The liver is an organ that plays a central role in nutrient metabolism, and when abnormal liver function causes problems in the metabolism of nutrients in the living body, it turns glucose into glycogen, converts protein into albumin, or breaks down unnecessary things and delivers them to bile juice. There is an abnormality in the function of the liver.

Among them, fatty liver is a disease that causes abnormal liver function due to abnormal accumulation of fat such as triglyceride in hepatocytes. It is known that the initial condition is simple fatty liver in which only fat deposition occurs in hepatocytes, and then the condition progresses to steatohepatitis including hepatic fibrosis, cirrhosis, and the like.

Fatty liver is classified into alcoholic fatty liver and non-alcoholic fatty liver according to its cause. Alcoholic fatty liver disease is thought to have progressed from initial simple fatty liver to fatty hepatitis and cirrhosis, while non-alcoholic fatty liver disease stays in simple fatty liver and the condition does not progress. It has been reported that the condition progresses due to steatohepatitis or cirrhosis.

Most patients with non-alcoholic fatty liver disease are accompanied by insulin resistance, obesity, diabetes and hyperlipidemia. In particular, 69-100% of non-alcoholic fatty liver patients are obese patients, and 20-40% of obese patients are accompanied by non-alcoholic fatty liver, and in particular, the prevalence of liver disease in male obese patients is higher than that of female obese patients. To date, there are two major treatments used for these patients. The first is a drug that treats and improves fatty liver through correction of risk factors, such as an obesity drug, an insulin resistance drug, or a hyperlipidemia drug, and the second is to recover damaged hepatocytes. It is a drug that is responsible for the function of letting go of liver cell protective agents, antioxidants, or nutritional support. However, until now, there is no effective drug treatment for non-alcoholic fatty liver disease, but only basic treatments such as weight loss through diet and exercise therapy are recommended. In particular, since non-alcoholic steatohepatitis is highly likely to develop into cirrhosis or hepatocellular carcinoma, more active drug treatment is essential, and improvement of oxidative stress and insulin resistance, which are considered important for the onset and progression of the condition of non-alcoholic steatohepatitis. Treatment aimed at is also being attempted, but the development of a therapeutic drug with high efficacy against non-alcoholic fatty liver disease is a reality because there is no treatment with sufficient scientific basis yet.

Therefore, according to the above demand, there is a need for the production of a product that is safe from plant extracts that have proven safety and is effective in preventing and improving metabolic diseases for effective prevention and improvement of the metabolic disease, which is rapidly increasing in incidence.

Meanwhile, Korean Patent Application Publication No. 2015-0027676 discloses a composition for preventing or treating metabolic syndrome-related diseases including grape and Schisandra chinensis extract, but the plant extract of the present invention, particularly bellflower extract, celery extract, and green tea extract. There is no disclosed composition for preventing, improving or treating metabolic diseases containing a mixture as an active ingredient.

The present invention provides a composition for preventing, improving or treating metabolic diseases containing a mixture of bellflower extract, celery extract, and green tea extract as an active ingredient, and the composition is a single extract or bellflower Compared to the mixture of the extract and the celery extract, the present invention was completed by confirming that there is an effect on improving obesity, hyperlipidemia, arteriosclerosis and cardiovascular disease, hyperinsulinemia, and fatty liver in a high fat diet animal model.

In order to solve the above problems, the present invention provides a health functional food composition for preventing or improving metabolic diseases containing a mixture of bellflower extract, celery extract, and green tea extract as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating metabolic diseases containing a mixture of bellflower extract, celery extract, and green tea extract as an active ingredient.

In addition, the present invention provides a feed additive for preventing or improving metabolic diseases in animals other than humans containing a mixture of bellflower extract, celery extract, and green tea extract as an active ingredient.

The present invention relates to a composition for preventing, improving or treating metabolic diseases containing a mixture of plant extracts as an active ingredient, and the mixture of bellflower extract, celery extract and green tea extract of the present invention is a single extract and bellflower in a high-fat diet animal model. Compared to the mixture of the extract and celery extract, the weight gain rate and the weight of liver tissue were reduced, the content of blood sugar and triglycerides was reduced, the content of insulin and leptin in the blood was excellent, and the effect of reducing the content of insulin and leptin in the blood was excellent. cholesterol), lowering the total cholesterol content, reducing the arteriosclerosis index and cardiovascular risk factors. In addition, it has an effect of reducing the fat content of liver tissue, increasing catalase activity in liver tissue, and increasing the content of GSH (glutathione), as well as reducing the size of adipose tissue around the epididymis.

1 is a check of the change in the content of blood sugar (A) and blood triglyceride (B) in a high-fat diet animal model ingesting the extract of the present invention. Control is a general diet group, HFD is a high fat diet group, PGE is a high fat diet and bellflower extract administration group, AGE is a high fat diet and celery extract administration group, GTE is a high fat diet and green tea extract administration group, and PA mixture is a high fat diet. It is a group administered with a mixture of bellflower extract and celery extract, and PAG mixture induces a high-fat diet, and a mixture of bellflower extract, celery extract, and green tea extract is administered. #, ### means that the value is significantly increased in HFD compared to Control, # is p<0.05, ### is p<0.001. *, **, *** means that the value of the PAG mixture is significantly reduced compared to the PGE, AGE, GTE and PA mixture, * is p<0.05, ** is p<0.01, *** is p <0.001.
Figure 2 shows the total cholesterol (A), HDL-cholesterol (B) and LDL-cholesterol (C) content in the blood of a high-fat diet animal model ingested the extract of the present invention. Control is a general diet group, HFD is a high fat diet group, PGE is a high fat diet and bellflower extract administration group, AGE is a high fat diet and celery extract administration group, GTE is a high fat diet and green tea extract administration group, and PA mixture is a high fat diet. It is a group administered with a mixture of bellflower extract and celery extract, and PAG mixture induces a high-fat diet, and a mixture of bellflower extract, celery extract, and green tea extract is administered. ### means that the value is significantly increased in HFD compared to Control, and ### is p<0.001. *, **, *** means that the value of the PAG mixture is significantly reduced compared to the PGE, AGE, GTE and PA mixture, * is p<0.05, ** is p<0.01, *** is p <0.001. ns means that there is no significant difference between the values of PGE, AGE, GTE and PA mixture and PAG mixture.
Figure 3 shows the arteriosclerosis index (A) and cardiovascular disease risk factors (B) of the high fat diet animal model ingested the extract of the present invention. Control is a general diet group, HFD is a high fat diet group, PGE is a high fat diet and bellflower extract administration group, AGE is a high fat diet and celery extract administration group, GTE is a high fat diet and green tea extract administration group, and PA mixture is a high fat diet. It is a group administered with a mixture of bellflower extract and celery extract, and PAG mixture induces a high-fat diet, and a mixture of bellflower extract, celery extract, and green tea extract is administered. ### means that the value is significantly increased in HFD compared to Control, and ### is p<0.001. *, *** means that the value of PAG mixture is significantly reduced compared to PGE, AGE, GTE and PA mixture, * is p<0.05, *** is p<0.001.
Figure 4 shows the blood insulin concentration (A) and leptin concentration (B) of a high fat diet animal model ingested the extract of the present invention. Control is a general diet group, HFD is a high fat diet group, PGE is a high fat diet and bellflower extract administration group, AGE is a high fat diet and celery extract administration group, GTE is a high fat diet and green tea extract administration group, and PA mixture is a high fat diet. It is a group administered with a mixture of bellflower extract and celery extract, and PAG mixture induces a high-fat diet, and a mixture of bellflower extract, celery extract, and green tea extract is administered. ### means that the value is significantly increased in HFD compared to Control, and ### is p<0.001. *, **, *** means that the value of the PAG mixture is significantly reduced compared to the PGE, AGE, GTE and PA mixture, * is p<0.05, ** is p<0.01, *** is p <0.001.
5 is a microscope confirming the histological changes of the liver in a high-fat diet animal model ingesting the extract of the present invention. Control is a general diet group, HFD is a high fat diet group, PGE is a high fat diet and bellflower extract administration group, AGE is a high fat diet and celery extract administration group, GTE is a high fat diet and green tea extract administration group, and PA mixture is a high fat diet. It is a group administered with a mixture of bellflower extract and celery extract, and PAG mixture induces a high-fat diet, and a mixture of bellflower extract, celery extract, and green tea extract is administered.
Figure 6 is a check of the catalase activity (A) and GSH concentration (B) in the liver tissue of the high fat diet animal model ingested the extract of the present invention. Control is a general diet group, HFD is a high fat diet group, PGE is a high fat diet and bellflower extract administration group, AGE is a high fat diet and celery extract administration group, GTE is a high fat diet and green tea extract administration group, and PA mixture is a high fat diet. It is a group administered with a mixture of bellflower extract and celery extract, and PAG mixture induces a high-fat diet, and a mixture of bellflower extract, celery extract, and green tea extract is administered. ### means that the value is significantly reduced in HFD compared to Control, and ### is p<0.001. *, **, *** means that the value of the PAG mixture increases significantly compared to the PGE, AGE, GTE and PA mixture, * is p<0.05, ** is p<0.01, *** is p <0.001.
7 is a microscope showing the histological changes of adipose tissue around the epididymis in a high-fat dietary animal model ingesting the extract of the present invention. Control is a general diet group, HFD is a high fat diet group, PGE is a high fat diet and bellflower extract administration group, AGE is a high fat diet and celery extract administration group, GTE is a high fat diet and green tea extract administration group, and PA mixture is a high fat diet. It is a group administered with a mixture of bellflower extract and celery extract, and PAG mixture induces a high-fat diet, and a mixture of bellflower extract, celery extract, and green tea extract is administered.

The present invention relates to a health functional food composition for preventing or improving metabolic diseases containing a mixture of bellflower extract, celery extract, and green tea extract as an active ingredient.

The metabolic disease refers to one or more diseases selected from the group consisting of hyperlipidemia, obesity, arteriosclerosis, cardiovascular disease, hyperinsulinemia, and non-alcoholic fatty liver disease, and the non-alcoholic fatty liver disease is simple fatty liver, nutritive fatty liver, starvation fatty liver, It is obese fatty liver, diabetic fatty liver, steatohepatitis, liver fibrosis or cirrhosis, but is not limited thereto.

In one embodiment of the present invention, the mixture may be a mixture of bellflower extract, celery extract, and green tea extract in a weight ratio of 0.5-2:0.5-2:0.5-2, preferably bellflower extract, celery extract, and green tea extract It is to mix the weight ratio of 1:1:1, but is not limited thereto.

In one embodiment of the present invention, the bellflower extract, celery extract and green tea extract _{may be extracted using water, C 1} to C ₄ lower alcohol or a mixture thereof as a solvent, preferably bellflower and celery extract The extraction is performed using ethanol as a solvent, and the green tea extract is extracted using water as a solvent, but the present invention is not limited thereto.

When using the health functional food composition of the present invention as a food additive, the health functional food composition may be added as it is or may be used with other foods or food ingredients, and may be appropriately used according to a conventional method. The amount of the active ingredient can be appropriately used depending on the purpose of use (prevention or improvement). In general, when preparing food or beverage, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the total raw material. However, in the case of long-term intake for the purpose of health, the amount may be less than the above range, and there is no problem in terms of safety, so the active ingredient may be used in an amount above the above range.

There is no particular limitation on the types of the health functional food. Examples of foods to which the health functional food composition can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea There are drinks, alcoholic beverages, and vitamin complexes, and all health foods in the usual sense are included.

In addition, the health functional food composition of the present invention may be prepared as a food, particularly a functional food. Functional foods of the present invention include ingredients that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when prepared as a drink, a natural carbohydrate or flavoring agent may be included as an additional ingredient in addition to the active ingredient. The natural carbohydrates are monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g., dextrins, cyclodextrins, etc.) or sugar alcohols (e.g. , Xylitol, sorbitol, erythritol, etc.). As the flavoring agent, natural flavoring agents (eg, taumatin, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be used.

In addition to the above health functional food composition, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonic acid It may further contain a carbonation agent used in beverages. Although the ratio of the ingredients to be added is not very important, it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food composition of the present invention.

In addition, the present invention relates to a pharmaceutical composition for preventing or treating metabolic diseases containing a mixture of bellflower extract, celery extract, and green tea extract as an active ingredient.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier in addition to the active ingredient, and such carriers are commonly used in formulation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, Calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals It includes, but is not limited to, oil and the like. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components.

The appropriate dosage of the pharmaceutical composition according to the present invention is formulated in various ways depending on factors such as the formulation method, the mode of administration, the patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and response sensitivity. Can be.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like.

The pharmaceutical composition of the present invention may be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, and methods of using a biological response modifier for suppression and treatment of metabolic diseases.

The concentration of the active ingredient contained in the composition of the present invention can be determined in consideration of the purpose of treatment, the condition of the patient, the required period, etc., and is not limited to a specific range of concentration.

The pharmaceutical composition of the present invention is prepared in a unit dosage form by formulating using a pharmaceutically acceptable carrier or excipient according to a method that can be easily carried out by a person having ordinary knowledge in the art. It can be manufactured by enclosing it in a dose container. At this time, the formulation may be prepared in any one formulation selected from injections, creams, sprays, ointments, warnings, lotions, liniment, pasta and cataplasma, and may additionally include a dispersant or a stabilizer.

In addition, the present invention relates to a feed additive for preventing or improving metabolic diseases in animals other than humans containing a mixture of bellflower extract, celery extract and green tea extract as an active ingredient.

The feed additive may be in the form of a high concentrate, powder, or granule containing a mixture of 20 to 90% by weight of bellflower extract, celery extract, and green tea extract.

The feed additives of the present invention include organic acids such as citric acid, humic acid, adipic acid, lactic acid, malic acid, and phosphates such as sodium phosphate, potassium phosphate, acidic pyrophosphate, and polyphosphate (polyphosphate), polyphenols, catechins, and alpha. -It may further contain at least one selected from natural antioxidants such as tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid.

Animal feed additives containing a mixture of bellflower extract, celery extract, and green tea extract of the present invention, and feed containing the same, as auxiliary ingredients, amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal enzymes, digestion and absorption It can be used with substances such as enhancers, growth accelerators, disease prevention agents, and the like.

The feed additive may be administered to an animal alone in combination with other feed additives in an edible carrier. In addition, the feed additive may be easily administered as a top dressing or directly mixed with animal feed or separately from feed, in a separate oral formulation, by injection or transdermally, or in combination with other ingredients. Typically, a single daily dose or divided daily doses may be used, as is well known in the art. When the feed additives are administered separately from animal feed, as is well known in the art, the dosage form of the extract can be prepared in an immediate release or sustained release formulation by combining them with a non-toxic pharmaceutically acceptable edible carrier. Such edible carriers can be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the dosage form of the extract may be a tablet, a capsule, a powder, a toroki or a sugar-containing tablet, or a top dressing in a microdispersible form. When a liquid carrier is used, it may be a soft gelatin capsule, or a dosage form of a syrup or liquid suspension, emulsion or solution. In addition, the dosage form may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like.

In addition, animal feed in which a mixture of bellflower extract, celery extract, and green tea extract is included as a feed additive may be any protein-containing organic grain flour commonly used to meet the dietary needs of animals. These protein-containing grains typically consist primarily of corn, soybean meal or corn/soybean meal mix. The feed additive may be immersed, sprayed or mixed and added to the animal feed. The veterinary composition or feed additive of the present invention can be applied to the diet of companion animals.

Hereinafter, the present invention will be described in more detail using Preparation Examples and Examples. These preparation examples and examples are only for describing the present invention in more detail, and it is obvious to those of ordinary skill in the art that the scope of the present invention is not limited thereto.

Materials and methods

1. Preparation of extract

Bellflower ( Platycodon grandiflorus ) roots were purchased at a special agricultural product store in Jinan-gun, Jeollabuk-do, chopped and dried at 40℃ for 16 hours. Dried bellflower roots (100g) were pulverized, added to 50% (v/v) ethanol (2,000 ml), extracted for 3 days, filtered using a 0.45 μm filter, and the filtrate was lyophilized and recovered and -20℃ It was used for experiments while being stored

Celery ( Apium graveolens ) purchased dried celery at Deokseong Celery Farm in Bongnim-dong, Gangseo-gu, Busan, crushed celery (100g), added it to 80% (v/v) ethanol (2,000 ㎖), extracted for 3 days, and then used a 0.45 µm filter. It was filtered using, and the filtrate was lyophilized and recovered, and stored at -20°C for use in the experiment.

Green tea extract powder (catechin content of 38% (w/w) or more, water extract) was purchased from Mirae Biotech Co., Ltd. Pocheon-si, Gyeonggi-do and stored at -20°C for experiments.

The mixture of bellflower extract and celery extract was prepared by mixing each extract in a weight ratio of 1:1 (PA mixture), and the mixture of bellflower extract, celery extract, and green tea extract was prepared in a weight ratio of 1:1:1 to each of the above extracts. It was prepared by mixing (PAG mixture).

2. Breeding and diet of experimental animals

As for the experimental animals, 4 weeks old C57BL/6 male mice (18-20g) were purchased from Orient Bio Co., Ltd., and they were adapted to the environment of the breeding room for 1 week. The temperature of the feedlot was maintained at 18~24℃ and the relative humidity was 50~60%, and the contrast was controlled at a 12-hour cycle (09:00~21:00).

Mice were in the general diet group (normal group, Control), high fat diet group (control group, HFD), high fat diet and bellflower extract administration group (PGE 200 mg/kg/day), high fat diet and celery extract administration group (AGE 200 mg/kg/day) , High-fat diet and green tea extract administration group (GTE 200mg/kg/day), inducing a high-fat diet, bellflower extract and celery extract administration group (PA mixture 200mg/kg/day), inducing a high-fat diet, bellflower extract, celery The experiment was conducted by dividing into 7 groups of extract and green tea extract mixture administration group (PAG mixture 200mg/kg/day), and 5 animals per each experimental group were randomly divided and bred for 16 weeks.

Each extract was administered orally once a day at a final concentration of 200mg/kg. D12450B (10% kcal fat) for general feed and D12492 (60% kcal fat) for high fat feed were purchased from Central Experimental Animal Co., Ltd. and used. High fat feed was fed while refrigerated at 4°C, and drinking water and feed were freely ingested. Feed intake and body weight were measured at weekly intervals. This study was conducted in accordance with the regulations of the Animal Experimental Ethics Committee of Jeonju University.

3. Blood glucose measurement, blood collection and storage

Blood glucose was fasted for 12 hours in the last 16 weeks of the experiment, and tail vein blood was collected and measured using a blood glucose meter (cuu-Check, Roche Diagnostics Korea Co., Ltd, Seoul, Korea), and for biochemical analysis. Blood to be used was anesthetized after measuring blood glucose, and blood was collected from the heart. The collected blood was centrifuged at 3,000 rpm at 4° C. for 15 minutes to separate the serum, and the separated serum was stored at -80° C. before analysis and used.

4. Biochemical analysis of blood

Serum triglyceride, total cholesterol, and HDL (High Density Lipoprotein)-cholesterol content were analyzed using a measurement kit (Asan), and LDL (Low Density Lipoprotein)-cholesterol content was determined by Friedewald et al. (1972, Clin Chem). It was calculated as in Equation 1 below.

[Equation 1]

(LDL-cholesterol)=[total cholesterol-(HDL-cholesterol)]-total triglycerides/5

In addition, arteriosclerosis index and cardiovascular risk factors were calculated as shown in Equations 2 and 3 below.

[Equation 2]

Atherosclerosis Index=[total cholesterol-(HDL-cholesterol)]/(HDL-cholesterol)

[Equation 3]

Cardiovascular risk factor = total cholesterol/(HDL-cholesterol)

5. Blood insulin and leptin concentration measurement

Serum insulin concentration and leptin concentration were determined by an insulin ELISA kit (ultra sensitive mouse insulin ELISA kit, Crysral Chem Inc., Elk Grove Village, IL, USA) and a leptin ELISA kit (mouse leptin Quantikine ELISA kit, R&D systems, Inc., Minneapolis, USA). MN, USA) was used to measure according to the manufacturer's protocol.

6. Liver tissue weighing, fat accumulation and biochemical analysis

In the last 16 weeks at the end of the experiment, the liver was fasted for 12 hours, the liver tissue was removed and the total weight was measured with a precision electronic balance, and then a part of the liver tissue was rapidly frozen (-40°C) for the later experiment. Some were washed with saline, dried, and fixed with 4% (v/v) paraformaldehyde (pH 7.4), and then made into a paraffin block through a series of processes.

The paraffin block was cut to a thickness of 5 μm and subjected to a deparaffin and hydration process, and then fat accumulation in liver tissue was stained with H&E and observed under a microscope (×100, Olympus, Tokyo, Japan).

GSH (glutathione) level in liver tissue was measured according to the manufacturer's protocol using a GSH assay kit (United States Biological., USA),

Liver catalase activity was measured using a commercially available assay kit (Cayman Chemical., USA) according to the manufacturer's protocol.

7. Observation of the size of fat around the epididymis

After the end of the experiment, adipose tissue around the epididymis was removed, washed with saline, dried, and fixed with 4% (v/v) paraformaldehyde (pH 7.4), and a paraffin block was prepared through a series of procedures. The paraffin block tissue was cut to a thickness of 5 μm, and the cut tissue section was stained with H&E after undergoing a deparaffin and hydration process, and the size of fat was confirmed with a microscope (×100, Olympus, Tokyo, Japan). .

Example 1. Weight gain and tissue weight measurement by administration of a mixture of plant extracts in a high fat diet animal model

The effect of the mixture of bellflower extract, celery extract, and green tea extract of the present invention on weight gain and tissues due to a high fat diet was confirmed.

As a result, as shown in Table 1, it was confirmed that the weight of the mouse was increased by a high fat diet, and a mixture of bellflower extract (PGE), celery extract (AGE), green tea extract (GTE), bellflower extract and celery extract (PA mixture) or a mixture of bellflower extract, celery extract, and green tea extract (PAG mixture) to reduce the weight gain. In particular, administration of a mixture of bellflower extract, celery extract, and green tea extract showed the greatest reduction in weight gain.

The weight of liver tissue increased by the high fat diet was also reduced by the administration of plant extracts, and the reduction effect was the best when the mixture of bellflower extract, celery extract, and green tea extract was administered.

group Weight(g) Acquisition weight (g) Liver weight (g) Before experiment After experiment Control 19.68±1.27 ^a 26.88±2.07 ^c 7.20±2.07 ^c 0.89±0.05 ^d HFD 19.54±0.79 ^a 44.00±1.93 ^a 24.46±1.93 ^a 2.06±0.14 ^a HFD+PGE 19.60±0.90 ^a 41.00±1.50 ^a 21.40±1.50 ^a 1.43±0.17 ^b HFD+AGE 19.46±1.02 ^a 40.83±1.89 ^a 21.37±1.89 ^a 1.62±0.35 ^b HFD+GTE 19.88±0.93 ^a 40.90±2.69 ^a 21.02±2.69 ^a 1.35±0.14 ^b HFD+PA mixture 20.13±0.57 ^a 41.73±2.11 ^a 21.60±2.11 ^a 1.50±0.09 ^b HFD+PAG mixture 18.90±0.80 ^a 36.07±1.25 ^b 17.17±1.25 ^b 1.08±0.11 ^c

In Table 1, the letters a to d mean that there is a significant difference from each other, and p<0.05.

Example 2. Biochemical analysis of blood by administration of a mixture of plant extracts in a high fat diet animal model

The effect of the mixture of bellflower extract, celery extract, and green tea extract of the present invention on blood sugar, blood triglyceride and cholesterol content in a high fat diet animal model was confirmed.

As a result, it was confirmed that the content of blood sugar and triglycerides increased by a high-fat diet decreased by administration of bellflower extract, celery extract, green tea extract, bellflower extract and a mixture of celery extract or a mixture of bellflower extract, celery extract, and green tea extract. In particular, when a mixture of three extracts was administered, a remarkable reduction effect was exhibited (FIG. 1).

In addition, the content of total cholesterol in the blood was decreased by administration of a mixture of bellflower extract, celery extract, green tea extract, bellflower extract and celery extract, or a mixture of bellflower extract, celery extract, and green tea extract. The reduction effect was the most excellent (Fig. 2A).

In addition, there was no significant change in the content of HDL-cholesterol in total cholesterol (FIG. 2B ), but the content of LDL-cholesterol was a mixture of bellflower extract, celery extract, green tea extract, bellflower extract and celery extract, or bellflower extract, celery extract, and It was reduced by the administration of the mixture of green tea extract, and a significant reduction effect was confirmed when the mixture of the three extracts was administered (Fig. 2C).

As a result of calculating the arteriosclerosis index and cardiovascular disease risk factors through Equations 2 and 3 using the measured total cholesterol, triglyceride and HDL-cholesterol levels, bellflower extract, celery extract, green tea extract, bellflower extract and celery extract The value was decreased by administration of the mixture or the mixture of bellflower extract, celery extract, and green tea extract, and the administration of the mixture of the three extracts showed a remarkable reduction effect (FIG. 3).

Example 3. Changes in blood insulin and leptin content by administration of a mixture of plant extracts in a high fat diet animal model

The effect of the mixture of bellflower extract, celery extract and green tea extract of the present invention on blood insulin and leptin content in a high fat diet animal model was confirmed.

Blood insulin content was increased by a high fat diet, and showed a tendency to decrease by administration of a mixture of bellflower extract, celery extract, green tea extract, bellflower extract and celery extract, or a mixture of bellflower extract, celery extract, and green tea extract. When the mixture of the species extract was administered, the most excellent insulin reducing effect was confirmed (FIG. 4A).

In addition, the content of leptin in blood was also increased by a high-fat diet, and showed a tendency to decrease by administration of a mixture of bellflower extract, celery extract, green tea extract, bellflower extract and celery extract, or a mixture of bellflower extract, celery extract, and green tea extract. , In particular, the best leptin reduction effect was confirmed when the mixture of the three extracts was administered (FIG. 4B).

Example 4. Pathological analysis of liver tissue by administration of a mixture of plant extracts in a high fat diet animal model

The effect of high fat diet and administration of plant extracts on liver tissue was analyzed pathologically.

As a result of observing the extracted liver tissue by H&E staining at 100 times, the increased fat content in the liver tissue by a high fat diet was found to be a mixture of bellflower extract, celery extract, green tea extract, bellflower extract and celery extract, or bellflower extract, celery extract, and It was confirmed that the reduction was confirmed by the administration of the mixture of green tea extract, and in particular, the effect of reducing the fat content was the most excellent when the mixture of the three extracts was administered, showing a form similar to that of the general diet group (FIG.

Example 5. Analysis of biochemical changes in liver tissue by administration of a mixture of plant extracts in a high fat diet animal model

Catalase enzyme activity was measured to determine the peroxidation reaction of the liver. Catalase enzyme activity decreased by a high fat diet was increased by administration of a mixture of bellflower extract, celery extract, green tea extract, bellflower extract and celery extract or a mixture of bellflower extract, celery extract and green tea extract, and in particular, administration of a mixture of three extracts The increase in the activity of the catalase enzyme was remarkable (Fig. 6A).

In addition, the concentration of GSH reduced by a high fat diet was increased by administration of a mixture of bellflower extract, celery extract, green tea extract, bellflower extract and celery extract or a mixture of bellflower extract, celery extract and green tea extract, and in particular, administration of a mixture of three extracts. Showed the most excellent increase effect (Fig. 6B).

Example 6. Analysis of changes in adipose tissue around epididymis by administration of a mixture of plant extracts in a high-fat dietary animal model

The effect of high fat diet and extract administration on the adipose tissue around the epididymis was confirmed. As a result, the size of adipocytes in adipose tissue enlarged by a high fat diet was reduced when administered bellflower extract, celery extract, green tea extract, bellflower extract and a mixture of celery extract or a mixture of bellflower extract, celery extract, and green tea extract. When the mixture of the species extract was administered, the reduction effect was the most excellent (FIG. 7).

Claims (4)

At least one metabolic disease selected from the group consisting of arteriosclerosis, cardiovascular disease and hyperinsulinemia containing a mixture of bellflower extract, celery extract, and green tea extract mixed in a weight ratio of 0.5-2:0.5-2:0.5-2 as active ingredients Health functional food composition for the prevention or improvement of. The method of claim 1, wherein the bellflower extract, celery extract, and green tea extract are _{extracted using water, C 1} ~ C ₄ lower alcohol or a mixture thereof as a solvent. Health functional food composition for preventing or improving at least one metabolic disease selected from the group consisting of. At least one metabolic disease selected from the group consisting of arteriosclerosis, cardiovascular disease and hyperinsulinemia containing a mixture of bellflower extract, celery extract, and green tea extract mixed in a weight ratio of 0.5-2:0.5-2:0.5-2 as active ingredients Pharmaceutical composition for the prophylaxis or treatment of. In the group consisting of arteriosclerosis, cardiovascular disease, and hyperinsulinemia of animals other than humans containing a mixture of bellflower extract, celery extract and green tea extract mixed in a weight ratio of 0.5-2:0.5-2:0.5-2 as active ingredients Feed additives for the prevention or improvement of one or more selected metabolic diseases.

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