KR20200045036A - 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, alleviating, or treating metabolic diseases, comprising a mixture of plant extracts as an active ingredient. The mixture of a bellflower extract, a celery extract and a green tea extract of the present invention has excellent effects of alleviating obesity, hyperlipidemia, arteriosclerosis and cardiovascular diseases, hyperinsulinemia and fatty liver in a high fat diet animal model, and thus the composition of the present invention, comprising the mixture of the bellflower extract, the celery extract and the green tea extract as an active ingredient may be useful as a therapeutic agent for metabolic diseases, a health functional food composition for preventing or alleviating metabolic diseases, or an animal feed material.

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.

Recently, the development of metabolic diseases (Metabolic Disease, Metabolic Syndrome), including various diseases such as obesity, hyperlipidemia, hypertension, arteriosclerosis, hyperinsulinemia, diabetes or liver disease, has rapidly increased according to changes in economic development and eating habits. These diseases may occur, but in general, they are most often associated with various symptoms while being closely related to each other.

Obesity is widely known to cause chronic diseases such as fatty liver, hypertension, diabetes, and cardiovascular diseases. As of 2007, 1.7 billion people, or about 25% of the world's population, are overweight (BMI> 25), and children 5 One out of every child falls into childhood obesity, and the number of childhood obesity is rapidly increasing, making it a serious social problem. Obesity treatments sold at home and abroad include generic drugs based on orlistat, which is approved by the U.S. FDA, but generic drugs that inhibit lipase action include gastrointestinal side effects such as fatty stool, gas production, and fat-soluble vitamin absorption. There is this.

Hyperlipidemia is a condition in which more fat-rich substances are present in the blood and accumulate on the walls of blood vessels and cause inflammation, resulting in cardiovascular disease. As the lipid component such as cholesterol increases in blood, the flow of blood becomes not smooth, and as the lipid components adhere to the artery wall, chronic inflammatory reactions and arteriosclerosis are induced, and the resulting thrombus causes cardiovascular or cerebral blood vessels. It blocks and causes myocardial infarction, stroke or stroke.

The arteriosclerosis decreases and hardens the elasticity of the arteries by accumulating fatty substances containing cholesterol, phospholipids, calcium, etc. in the vascular lining, and the blood supply is inhibited or pressure increases due to the narrowed arteries, causing rupture, exfoliation, etc. It says the state in which it happens.

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

Hyperinsulinemia is a condition with high levels of insulin in the blood, and is a disease associated with obesity or diabetes, such as enhancing sympathetic nerve activity or promoting absorption of sodium by the kidneys. There is an urgent need for the development of therapeutic agents that can lower levels.

The liver is an organ that plays a central role in the metabolism of nutrients, and when abnormalities in liver function cause problems in nutrient metabolism in the living body, it makes glucose into glycogen, converts proteins into albumin, breaks down unnecessary ones, and delivers them to bile bile, etc. There is an abnormality in the function of the liver.

Among them, fatty liver is a disease in which fat such as triglyceride accumulates abnormally in liver cells, leading to abnormal liver function. It is known that the initial condition is a simple fatty liver in which only fat deposits occur in the liver cells, and thereafter, the condition progresses to fatty hepatitis including liver fibrosis, cirrhosis, and the like.

Fatty liver is classified into alcoholic fatty liver and non-alcoholic fatty liver according to the cause of occurrence. Alcoholic fatty liver disease progresses from simple fatty liver in the early stage to fatty hepatitis and cirrhosis, while non-alcoholic fatty liver disease is thought to remain in the simple fatty liver and does not progress, but recently, in the case of nonalcoholic fatty liver disease, even from simple fatty liver disease It has been reported that the condition may progress due to fatty hepatitis or cirrhosis.

Most patients with non-alcoholic fatty liver disease are accompanied by insulin resistance, obesity, diabetes and hyperlipidemia. In particular, 69 to 100% of non-alcoholic fatty liver patients are obese patients, and 20 to 40% of obese patients are accompanied by non-alcoholic fatty liver. In particular, the prevalence of liver disease in male obese patients is higher than that in female obese patients. To date, there are two therapeutic agents used in these patients: the first is a drug that treats and improves the fatty liver through correction of risk factors, such as an obesity drug, an insulin resistance drug, or a hyperlipidemic drug, and the second recovers damaged liver cells. Hepatocyte protection agents, antioxidants, or nutritional support are the drugs responsible for the function of prescribing. However, until now, there is no effective drug treatment for non-alcoholic fatty liver disease, but only basic treatment methods such as weight loss through diet and exercise therapy are recommended. In particular, since non-alcoholic steatohepatitis is more likely to develop into cirrhosis or hepatocellular carcinoma, more aggressive drug treatment is essential, and improvement of oxidative stress or insulin resistance, which is thought to be important for the pathogenesis and progression of nonalcoholic steatohepatitis, etc. Although treatment aiming at is also being attempted, there is no sufficient treatment to establish a sufficient scientific basis, so there is a need to develop a therapeutic drug with high effectiveness against non-alcoholic fatty liver disease.

Accordingly, there is a need for production of products that are safe and effective in the prevention and improvement of metabolic diseases from plant extracts proven to be safe for the effective prevention and improvement of the metabolic diseases in which the onset is rapidly increasing.

On the other hand, Korean Patent Publication No. 2015-0027676 discloses a composition for the prevention or treatment of metabolic syndrome-related diseases, including grape and Omija complex extracts, but the plant extracts of the present invention, in particular bellflower extract, celery extract and green tea extract No composition has been disclosed for preventing, improving or treating metabolic diseases containing the mixture as an active ingredient.

The present invention has been derived by the above-described needs, 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, wherein the composition is a single extract or bellflower The present invention was completed by confirming that there is an effect of improving obesity, hyperlipidemia, arteriosclerosis and cardiovascular disease, hyperinsulinemia and fatty liver in a high fat diet animal model compared to a mixture of extract and celery extract.

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 the prevention or improvement of 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, 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 extract and celery extract, the weight gain rate and the weight of liver tissue are reduced, the content of blood sugar and triglycerides is reduced, and the effect of reducing the content of insulin and leptin in blood is excellent, and LDL-cholesterol (low density lipoprotein) in blood cholesterol) to lower the total cholesterol content, and to reduce the risk factors of arteriosclerosis and cardiovascular disease. In addition, the effect of reducing the fat content of the liver tissue, increasing the catalase activity in the liver tissue, and increasing the content of GSH (glutathione), as well as the effect of reducing the size of the fat tissue around the epididymis.

Figure 1 confirms the change in the content of blood glucose (A) and 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 group. Induction, the mixture administration group of bellflower extract and celery extract, PAG mixture induces a high fat diet, and is a group administered with a mixture of bellflower extract, celery extract and green tea extract. #, ### means that the value increases significantly in HFD compared to Control, # is p <0.05, and ### is p <0.001. *, **, *** mean that the value of PAG mixture is significantly reduced compared to 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 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 group. Induction, the mixture administration group of bellflower extract and celery extract, PAG mixture induces a high fat diet, and is a group administered with a mixture of bellflower extract, celery extract and green tea extract. ### means that the value increases significantly in HFD compared to Control, and ### is p <0.001. *, **, *** mean that the value of PAG mixture is significantly reduced compared to 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 in PGE, AGE, GTE and PA mixture and PAG mixture values.
Figure 3 shows the arteriosclerosis index (A) and cardiovascular disease risk factors (B) of the high-fat diet animal model ingested with 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 group. Induction, the mixture administration group of bellflower extract and celery extract, PAG mixture induces a high fat diet, and is a group administered with a mixture of bellflower extract, celery extract and green tea extract. ### means that the value increases significantly 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 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 group. Induction, the mixture administration group of bellflower extract and celery extract, PAG mixture induces a high fat diet, and is a group administered with a mixture of bellflower extract, celery extract and green tea extract. ### means that the value increases significantly in HFD compared to Control, and ### is p <0.001. *, **, *** mean that the value of PAG mixture is significantly reduced compared to PGE, AGE, GTE and PA mixture, * is p <0.05, ** is p <0.01, *** is p <0.001.
5 is a microscopic examination of histological changes in 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 group. Induction, the mixture administration group of bellflower extract and celery extract, PAG mixture induces a high fat diet, and is a group administered with a mixture of bellflower extract, celery extract and green tea extract.
Figure 6 confirms 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 group. Induction, the mixture administration group of bellflower extract and celery extract, PAG mixture induces a high fat diet, and is a group administered with a mixture of bellflower extract, celery extract and green tea extract. ### means that the value is significantly decreased in HFD compared to Control, and ### is p <0.001. *, **, *** mean that the value of PAG mixture increases significantly compared to PGE, AGE, GTE and PA mixture, * means p <0.05, ** means p <0.01, *** means p <0.001.
7 is a microscopic examination of the histological changes of adipose tissue surrounding the epididymis 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 group. Induction, the mixture administration group of bellflower extract and celery extract, PAG mixture induces a high fat diet, and is a group administered with a mixture of bellflower extract, celery extract and green tea extract.

The present invention relates to a health functional food composition for preventing or improving metabolic disease 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, nutrient fatty liver, starvation fatty liver, Obese fatty liver, diabetic fatty liver, fatty hepatitis, 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 to 2: 0.5 to 2: 0.5 to 2, preferably bellflower extract, celery extract and green tea extract. It is to be mixed in a 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, a lower alcohol of C ₁ to C ₄ or a mixture thereof as a solvent, preferably the bellflower and celery extract Ethanol is extracted using a solvent, and green tea extract is extracted using water as a solvent, but is not limited thereto.

When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is or used with other foods or food ingredients, and may be suitably 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, the health functional food composition of the present invention in the manufacture of a food or beverage is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less with respect to the total raw material. However, in the case of long-term intake for health purposes, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

There are no particular restrictions on the type 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 products, dairy products including ice cream, various soups, beverages, tea Drinks, alcoholic beverages, and vitamin complexes are included, and include all healthy foods in the ordinary sense.

In addition, the health functional food composition of the present invention may be prepared as a food, particularly a functional food. The functional food of the present invention includes ingredients that are commonly added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when prepared as a drink agent, natural carbohydrate or flavoring agent may be included as an additional component in addition to the active ingredient. The natural carbohydrates include 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, and the like). The flavoring agent may be a natural flavoring agent (eg, tau martin, stevia extract, etc.) and a synthetic flavoring agent (eg, saccharin, aspartame, etc.).

Various nutritional supplements, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonic acid It may further contain a carbonation agent used in the beverage. The proportion of the components to be added is not particularly important, but it is generally selected from the range of 0.01 to 0.1 parts by weight with respect to 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 a carrier is commonly used in preparation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, Calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals Oil, and the like, but is not limited thereto. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components.

Suitable dosages of the pharmaceutical compositions according to the invention are variously prescribed by factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, and response sensitivity. Can be.

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

The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers for the inhibition 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 treatment purpose, the patient's condition, and the required period, 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 skilled in the art to which the present invention pertains, or It can be prepared by incorporating into a dose container. At this time, the formulation may be prepared as any one of a formulation selected from injections, creams, sprays, ointments, warning agents, lotions, linen agents, pasta agents, and cataplasmas, and may further 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 20-90% 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, and malic acid, or phosphates such as sodium phosphate, potassium phosphate, acidic pyrophosphate, polyphosphate (polymerized phosphate), polyphenols, catechin, alpha -Tocopherol, rosemary extract (rosemary extract), vitamin C, green tea extract, licorice extract, chitosan, tannic acid, may further include one or more selected from natural antioxidants such as phytic acid.

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

The feed additive may be administered to animals alone in combination with other feed additives in an edible carrier. In addition, the feed additive can be easily administered as a top dressing or by mixing them directly into animal feed or separately from the feed, in a separate oral dosage form, by injection or transdermal or in combination with other ingredients. Typically, a single daily dose or a divided daily dose can be used, as is well known in the art. When the feed additives are administered separately from the animal feed, the dosage form of the extract, as is well known in the art, 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, such as 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 can be a tablet, capsule, powder, torokis or sugar-coated tablets or top dressing in undispersed form. When a liquid carrier is used, it may be in the form of a soft gelatin capsule, or a syrup or liquid suspension, emulsion or solution. In addition, dosage forms may contain adjuvants, such as preservatives, stabilizers, wetting or emulsifying agents, solution accelerators, and the like.

In addition, the animal feed in which the mixture of bellflower extract, celery extract and green tea extract is included as a feed additive can be any protein-containing organic flour commonly used to meet the animal's dietary needs. These protein-containing flours usually consist mainly of corn, soy flour or corn / soy flour mixtures. The feed additive may be used by adding to the animal feed by dipping, spraying or mixing. 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 by using manufacturing examples and examples. It is obvious to those skilled in the art that these production examples and examples are only intended to illustrate the present invention more specifically and that the scope of the present invention is not limited by them.

Materials and methods

1. Preparation of Extract

The roots of Platycodon grandiflorus were purchased from a specialty agricultural product store in Jinan-gun, Jeollabuk-do, and then chopped and dried at 40 ℃ for 16 hours. The dried bellflower root (100 g) was crushed, 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 then recovered and recovered at -20 ° C. It was used for the experiment while being stored at.

Celium ( Apium graveolens ) purchased dried celery at Deokseong celery farm in Bongrim-dong, Gangseo-gu, Busan, crushed celery (100 g), added to 80% (v / v) ethanol (2,000 mL), extracted for 3 days, and then filtered for a 0.45 μm filter Filtrate was used, and the filtrate was lyophilized and then 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 was used in the experiment while storing at -20 ℃.

A mixture of bellflower extract and celery extract was prepared by mixing each extract in a weight ratio of 1: 1 (PA mixture), and a mixture of bellflower extract, celery extract, and green tea extract weighted each extract as 1: 1: 1 It was prepared by mixing (PAG mixture).

2. Experimental animal  Breeding and diet

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

The mice were the normal diet group (normal group, control), high fat diet group (control group, HFD), high fat diet and bellflower extract administration group (PGE 200mg / kg / day), high fat diet and celery extract administration group (AGE 200mg / kg / day). , High fat diet and green tea extract administration group (GTE 200mg / kg / day), induces high fat diet, and a mixture administration group of bellflower extract and celery extract (PA mixture 200mg / kg / day), high fat diet, induces bellflower extract, celery The experiment was conducted by dividing into a total of 7 groups of the mixture administration group of the extract and green tea extract (PAG mixture 200mg / kg / day), and 5 animals were randomly divided for each experiment group and kept for 16 weeks.

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

3. Blood sugar measurement, blood collection and storage

Blood glucose was fasted for 12 hours in the last 16 weeks of the end 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). The blood to be used was anesthetized after blood glucose measurement and then taken from the heart. The collected blood was centrifuged at 4 ° C and 3,000 rpm for 15 minutes to separate the serum, and the separated serum was used while being stored at -80 ° C until analysis.

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 measured by Friedewald et al. (1972, Clin Chem). It was calculated as shown in Equation 1 below.

[Equation 1]

(LDL-Cholesterol) = [Total cholesterol- (HDL-Cholesterol)]-Total triglyceride / 5

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

[Equation 2]

Arteriosclerosis Index = [Total Cholesterol- (HDL-Cholesterol)] / (HDL-Cholesterol)

[Equation 3]

Risk factors for cardiovascular disease = total cholesterol / (HDL-cholesterol)

5. Blood insulin and Leptin  Concentration measurement

Serum insulin concentration and leptin concentration are insulin sensitive ELISA kit (ultra sensitive mouse insulin ELISA kit, Crysral Chem Inc., Elk Grove Village, IL, USA) and leptin ELISA kit (mouse leptin Quantikine ELISA kit, R & D systems, Inc., Minneapolis, MN, USA).

6. Liver tissue weighing, fat accumulation and biochemical analysis

Fasting for 12 hours in the last 16 weeks of the end of the experiment, extracting liver tissue, measuring the total weight with a precision electronic balance, and then rapidly freezing (-40 ° C) part of the liver tissue for further experiments. After washing with some brine, some of the water was removed and fixed with 4% (v / v) paraformaldehyde (pH 7.4) to produce a paraffin block through a series of processes.

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

GSH (glutathione) levels in liver tissue were 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. Observing the size of fat around the epididymis

After the end of the experiment, the adipose tissue around the epididymis was extracted, washed with saline, and then drained, fixed with 4% (v / v) paraformaldehyde (pH 7.4) to prepare a paraffin block through a series of processes. The paraffin block tissue was cut to a thickness of 5 μm, and the cut tissue sections were dehydrated with deparaffin and stained with H & E to confirm the size of fat under 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 tissue by high fat diet was confirmed.

As a result, as shown in Table 1, it was confirmed that the weight of the mouse 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. Particularly, when the mixture of bellflower extract, celery extract and green tea extract was administered, the effect of reducing the weight gain was the best.

The weight of liver tissue increased by a high fat diet was also decreased 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 the experiment After the 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

The letters a to d in the column 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 the blood glucose, triglyceride and cholesterol content of the 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 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, when the mixture of the three extracts was administered, it showed a significant reduction effect (Fig. 1).

In addition, the content of total cholesterol in the blood was reduced 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, particularly by administration of a mixture of three extracts. The reduction effect was the best (Fig. 2A).

In addition, the content of HDL-cholesterol in total cholesterol did not change significantly (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 administration of the mixture of green tea extract, and a significant reduction effect was observed when the mixture of the three extracts was administered (FIG. 2C).

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

Example  3. In the high fat diet animal model, blood insulin and Leptin  Content change

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

The insulin content in the blood 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, particularly 3 When the mixture of the species extract was administered, the best insulin reduction effect was confirmed (FIG. 4A).

In addition, the content of leptin in the 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, when the mixture of the three extracts was administered, the best leptin reduction effect was confirmed (Fig.

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 plant extract administration 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 high fat diet is 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 by administration of the mixture of the green tea extract, in particular, when the mixture of the three extracts showed the best effect of reducing the fat content, it showed a form similar to that of the general diet (Fig. 5).

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 investigate the liver peroxidation reaction. The catalase enzyme activity reduced by the 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, particularly administering a mixture of three extracts Cycatalase enzyme activity was remarkable increase (Fig. 6A).

In addition, the GSH concentration 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, particularly administering a mixture of three extracts Showed the best increase effect (Fig. 6B).

Example  6. Analysis of changes in adipose tissue surrounding the epididymis by administration of a mixture of plant extracts in a high fat diet animal model

The effect of high fat diet and extract administration on the fat tissue around the epididymis was confirmed. As a result, the size of fat cells in the adipose tissue enlarged by a high fat diet was reduced upon 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, 3 When the mixture of species extract was administered, the reduction effect was the best (FIG. 7).

Claims (7)

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 method of claim 1, wherein the metabolic disease is hyperlipidemia, obesity, arteriosclerosis, cardiovascular disease, hyperinsulinemia and non-alcoholic fatty liver disease, one or more selected from the group consisting of health functional food composition for preventing or improving metabolic disease . The metabolic property of claim 2, wherein the non-alcoholic fatty liver disease is any one selected from the group consisting of simple fatty liver, nutrient fatty liver, hunger fatty liver, obese fatty liver, diabetic fatty liver, fatty hepatitis, liver fibrosis and cirrhosis. Health functional food composition for preventing or improving disease. According to claim 1, The mixture is a bellflower extract, celery extract and green tea extract 0.5-2: 0.5-2: 0.5-2 health functional food composition for preventing or improving metabolic disease characterized in that it is mixed in a weight ratio. The method of claim 1, wherein the bellflower extract, celery extract and green tea extract is water, C ₁ ~ C ₄ lower alcohol or a mixture thereof as a solvent, characterized in that extracted using a solvent for preventing or improving metabolic disease health functional food Composition. A pharmaceutical composition for the prevention or treatment of metabolic diseases containing a mixture of bellflower extract, celery extract and green tea extract as an active ingredient. A feed additive for preventing or improving metabolic diseases in animals except humans containing a mixture of bellflower extract, celery extract and green tea extract as an active ingredient.

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