KR20190058110A - Use of astragalus membranaceus, rubus coreanus and helianthus tuberosus extract mixture for treating diabetes - Google Patents

Abstract

The present invention relates to the anti-diabetic use of a combination of hwanggi, bokbunja and porcini potato complex extracts, wherein a specific mixture of hwanggi, bokbunja, and porcine potato extracts mixed with a specific ratio of the present invention shows glucose production without toxicity in high glucose- , Reduce the fasting blood glucose level in the diabetic model mouse, decrease the glycated hemoglobin, and reduce the oral glucose tolerance, and thus can be used for prevention and treatment of diabetes through the hypoglycemic effect .

Description

[0001] The present invention relates to an antidiabetic agent for use in combination with an antidiabetic agent,

The present invention relates to antidiabetic uses of a combination of hwanggi, bokbunja and porcini potato complex.

Diabetes is one of the most common chronic diseases, and the number of patients is rapidly growing due to the improvement of living standards and the westernized lifestyle. The control of the body in our body is made by the balance of the consumption of the party and the production of sugar. Among them, insulin, glucagon and others are known. These hormones play a role in maintaining blood sugar within a certain range. If abnormalities of these hormones occur, problems arise in the regulation of the blood sugar level and diabetes is a typical disease (Shepherd PR, Kahn BB: Glucose transporters and insulin action-implications for insulin resistance and diabetes mellitus. N Engl J Med 341: 248-257, 1999). Abnormal glucose levels due to diabetes increase the level of hemoglobin in the plasma and are associated with chronic hyperglycemia, atherosclerosis, microangiopathy, diabetic neuropathy, kidney disease, heart disease , Diabetic retinopathy, and cataracts, leading to death (American Diabetes Association: Diagnosis and classification of diabetes mellitus. Diabetes Care 27: S5-S10, 2004).

Diabetes mellitus is characterized by two types: insulin dependent diabetes (IDDM), which is caused by insulin secretion deficiency in the blood, the glucose regulating hormone in the blood, It is also called juvenile diabetes because it occurs in younger age groups. Type II diabetes (non-insulin dependent diabetes, NIDDM) occurs mainly after the age of 40, and it accounts for most of the diabetic patients in Korea. Unlike type Ⅰ, it is called adult type diabetes. The cause of the disease is not clear yet, but it is known that genetic factors and environmental factors are involved together. As a pathogenesis of type II diabetes, both insulin secretion defects in pancreatic beta cells and insulin action defects in target cells (insulin resistance) are observed, although it is not yet clear which change is of primary importance. Currently, there are five classes of compounds used for complications such as type 2 diabetes and insulin resistance: biguanides, thiazolidinediones, sulfonylureas, benzoic acid, Derivative compounds and? -Glucosidase inhibitors have been used. Of these, it has been known that acetylated compounds such as metformin prevent excessive gluconeogenesis in the blood. Thiazolidinedione compounds are believed to act to increase the rate of glucose consumption of the peripheral nerves and include compounds such as sulfonylureas, such as tolbutamide and glyburide, benzoic acid, such as repaglinide, Derivatives and alpha glucosidase inhibitors such as acarbose act to lower plasma glucose by stimulating insulin secretion.

Control of blood sugar in the treatment of diabetes is one of the most important goals. Controlling postprandial blood glucose as well as fasting blood glucose as close to normal as possible is very important in the treatment and prevention of diabetes (Jenkins, DJA et al., Diabetes Care, 11, pp. 149-159, 1988; Harold, E., Diabetic Research abd Clinical Practice, 40, ppS27-S28, 1998; Hermann, H., Diabetic Research abd Clinical Practice, 40, ppS43-S49, al., Diabetes Research and Clinical Practice, 43 (3), pp 179-185, 1999). However, the above sulfonylurea and biguanide-containing preparations have various drawbacks and side effects, and therefore, there is a need for an excellent hypoglycemic agent having low side effects and high safety.

Accordingly, the present inventors have completed the present invention by confirming the excellent hypoglycemic effect of a combination of Hwanggi, Bokbunja and Pork Potato Extract while studying to provide a natural substance excellent in blood glucose lowering action without adverse effects on the living body

In the present invention, it is an object of the present invention to use them as a composition for prevention or treatment of diabetes and a food composition for lowering blood glucose by confirming the antidiabetic activity, particularly the hypoglycemic function, of a mixture of extracted yellowness, bacterium and porcine potato mixed at a specific ratio.

In order to achieve the above object, the present invention provides a composition for preventing or treating diabetes mellitus containing an extract of hwanggi, bokbunja, and porcine potato as an active ingredient.

The present invention also provides a food composition for hypoglycemia containing an extract of hwanggi, bokbunja and pig potato as an active ingredient.

In addition, the present invention provides a method for preparing an extract of hwanggi, bokbunja, and porcine potatoes showing antidiabetic and hypoglycemic effects.

According to the present invention, the specific mixture of yellow, blue, and pig potato extracts mixed at a specific ratio inhibits sugar production without showing toxicity in high glucose-induced hepatocytes, decreases fasting blood glucose in diabetic model mice, decreases glycosylated hemoglobin And reduces oral glucose tolerance. Thus, it can be used for prevention and treatment of diabetes through blood glucose lowering effect.

FIG. 1 shows cell viability of HepG2 hepatocyte induced by high glucose. FIG.
FIG. 2 is a graph showing the ability of hepG2 hepatocyte-induced hepatic glucose production to inhibit glucose production.
FIG. 3 is a graph showing inhibition of sugar production by mixing ratio of hwanggi, bokbunja and porcine potatoes in high glucose-induced HepG2 hepatocytes.
FIG. 4 is a graph showing the cell survival rate according to the mixing ratio of hwanggi, bokbunja and porcine potatoes in HepG2 hepatocyte-induced hepatocyte.
FIG. 5 is a graph showing the expression levels of proteins known to be involved in sugar production.
6 is a chart for confirming fasting blood glucose in an animal model in which diabetes is induced.
FIG. 7 is a chart showing glycated hemoglobin in an animal model in which diabetes is induced.
FIG. 8 is a chart for confirming oral glucose load in a diabetic-induced animal model. FIG.

Hereinafter, the present invention will be described in detail with reference to embodiments of the present invention. It should be understood, however, that the invention is not limited thereto and that various changes and modifications may be made therein without departing from the spirit and scope of the invention as defined by the appended claims and their equivalents. .

In one aspect, the present invention relates to a composition for the prevention or treatment of diabetes comprising an extract of hwanggi, bokbunja and potato extract as an active ingredient.

In one embodiment, the Yellow, Blue, and Piglet potato extract complexes can be mixed in a ratio of 3: 1: 2 (w / w) of Hwanggi, Bokbunja and Pork potato extract.

In one embodiment, the extract of hwanggi, bokbunja and pig potato can be extracted with water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, and more preferably, it is extracted with hot water.

The term " extract " used in the present invention means a preparation which is obtained by squeezing a herbal medicine with an appropriate leaching solution and concentrating the liquid by evaporating the leaching solution. The extract is not limited thereto, but may be a diluent or concentrate of the extract, A dried product obtained by drying the extract, a controlled preparation thereof, or a purified product thereof. The extract can be prepared by a general extraction method, separation and purification methods known in the art. The extraction method may be, but not limited to, hot water extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction.

The term " complex extract " or " extracted complex " as used in the present invention refers to both complex extracts obtained by extracting and mixing together hwanggi, bokbunja and porcine potato, or complex extracts obtained by extracting hwanggi,

In the present invention, the extract may be prepared by extracting with an extraction solvent or extracting with an extraction solvent, followed by fractionation with a fraction solvent. The organic solvent may be an alcohol having 1 to 4 carbon atoms, a polar solvent such as ethyl acetate or acetone, a solvent such as hexane or dichloro, or an organic solvent such as dichloromethane, Methane, or a mixed solvent thereof may be used. Further, water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof may be preferably used, and hot water may be more preferably used. In one embodiment of the present invention, each of the extracts was subjected to hot-water extraction with reflux using tertiary distilled water as the solvent, followed by concentration under reduced pressure.

In one embodiment, the yellow moss, hyssop, and pig potato extract complexes can be administered at a concentration of 10 to 200 mg / kg, more preferably 50 to 100 mg / kg.

In the present invention, the term " prevention " means any action that inhibits or delays the generation, spread and recurrence of diabetes by the administration of the pharmaceutical composition according to the present invention.

As used herein, the term " treatment " means any action that improves or alleviates the symptoms of diabetes or diabetic complications by administration of a composition comprising the extract of hwanggi, bokbunja, and porcine potato extract of the present invention. Those skilled in the art will be able to ascertain the precise criteria of the disease for which the composition of the present invention is effective by referring to the data presented by the Korean Medical Association, will be.

The term " therapeutically effective amount " used in connection with the active ingredient in the present invention means the amount of a pharmaceutically acceptable salt of the extract of the invention which is effective for preventing or treating the target disease, The amount can vary depending on a number of factors, such as the mode of administration, the site of administration, the condition of the patient, and the like. Therefore, when used in the human body, the dosage should be determined in consideration of safety and efficacy. It is also possible to estimate the amount used in humans from the effective amount determined through animal experiments. Such considerations in determining the effective amount are described, for example, in Hardman and Limbird, eds., Goodman and Gilman ' s Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; And E.W. Martin ed., Remington ' s Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term " pharmaceutically effective amount " means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, Factors well known in the art and other medical disciplines including health status, type of cancer, severity, activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration and rate of release, duration of treatment, ≪ / RTI > The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The pharmaceutical compositions of the present invention may include carriers, diluents, excipients, or a combination of two or more thereof commonly used in biological formulations. As used herein, the term " pharmaceutically acceptable " means that the composition is free of toxicity to cells or humans exposed to the composition. The carrier is not particularly limited as long as the composition is suitable for in vivo delivery, for example, Merck Index, 13th ed., Merck & Inc. A buffered saline solution, a buffer solution, a dextrose solution, a maltodextrin solution, glycerol, ethanol, and one or more of these components may be mixed and used, and if necessary, an antioxidant, a buffer, Conventional additives may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into main dosage forms such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules or tablets. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990) in a suitable manner in the art.

The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, Wherein the starch is selected from the group consisting of lactose, mannitol, sugar, arabic gum, pregelatinized starch, cornstarch, powdered cellulose, hydroxypropyl cellulose, opaques, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, Calcium, white sugar, dextrose, sorbitol and talc may be used. The pharmaceutically acceptable additive according to the present invention is preferably included in the composition in an amount of 0.1 part by weight to 90 parts by weight, but is not limited thereto.

The pharmaceutical composition of the present invention may further include a known diabetes therapeutic agent in addition to the yellow, bramble and pig potato extract complexes as active ingredients, and may be used in combination with other treatments known for the treatment of these diseases.

The term " administering " as used herein means providing a predetermined substance to a patient in any appropriate manner, and may be administered orally or parenterally (for example, by intravenous, subcutaneous, The dosage may vary depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease. Examples of the liquid preparation for oral administration of the composition of the present invention include suspensions, solutions, emulsions, syrups, and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, Etc. may be included together. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. The pharmaceutical composition of the present invention may be administered by any device capable of moving the active substance to the target cell. The preferred modes of administration and formulations are intravenous, subcutaneous, intradermal, intramuscular, and drip injections. The injectable solution may be a non-aqueous solvent such as an aqueous solvent such as a physiological saline solution or a ring gel solution, a vegetable oil, a higher fatty acid ester (e.g., oleic acid), an alcohol (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.) (For example, ascorbic acid, sodium hydrogen sulfite, sodium pyrophosphate, BHA, tocopherol, EDTA and the like), an emulsifier, a buffer for pH control, a microbial growth inhibitor And a pharmaceutical carrier such as a preservative (e.g., mercury nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

In one aspect, the present invention relates to a food composition for hypoglycemia containing as an active ingredient an extract of yellow mosses, brambles and potatoes.

In one embodiment, the yellow, bellwether, and porcine potato extract complexes can be mixed in a ratio of 3: 1: 2 (w / w) of hwanggi, bokbunja and porcini potato extracts, have.

When the composition of the present invention is used as a food composition, it is possible to add the above-described hwanggi, bokbunja and porcini potato extract complexes directly or in combination with other food or food ingredients and suitably use according to a conventional method. The composition may contain a food-acceptable food-aid additive in addition to the active ingredient, and the amount of the active ingredient to be mixed may be suitably determined according to the intended use (prevention, health or therapeutic treatment).

As used herein, the term " food-aid additive " refers to a component that can be added to foods in a supplementary manner, and is appropriately selected and used by those skilled in the art as added to produce health functional foods of each formulation. Examples of food-aid additives include flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, , a pH adjusting agent, a stabilizer, a preservative, a glycerin, an alcohol, and a carbonating agent used in a carbonated drink. However, the types of the food auxiliary additives of the present invention are not limited by these examples.

A health functional food may be included in the food composition of the present invention. The term " health functional food " as used in the present invention refers to a food prepared and processed in the form of tablets, capsules, powders, granules, liquids and rings using raw materials and components having useful functions in the human body. Here, 'functional' refers to the structure and function of the human body to obtain nutritional effects and obtain useful effects for health use such as physiological action. The health functional food of the present invention can be prepared by a method commonly used in the art, and can be prepared by adding raw materials and components which are usually added in the conventional technical fields. In addition, the formulations of the above health functional foods may also be manufactured without limitations as long as they are acceptable as health functional foods. The composition for food of the present invention can be manufactured in various forms, and unlike general pharmaceuticals, it has the advantage that there is no side effect that may occur when a drug is used for a long period of time, and is excellent in portability, Can be ingested as an adjuvant to enhance the effectiveness of anticancer drugs.

There is no limitation on the kind of health food to which the composition of the present invention can be used. In addition, the composition comprising the hwanggi, bokbunja, and porcini potato extract composite of the present invention as an active ingredient may be prepared by mixing other suitable auxiliary ingredients and known additives, which may be contained in health functional foods, according to the selection of a person skilled in the art. Examples of foods that can be added include dairy products, such as meat, sausage, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Vitamin complex, and the like, and can be prepared by adding to the juice, tea, jelly, and juice prepared from the extract of the present invention as a main component.

Since the extract of hwanggi, bokbunja and porcine potato extract of the present invention uses natural plant as a raw material, even when it is used as a pharmaceutical composition or a food composition, side effects can be less than general synthetic compounds, and therefore they are safely contained in pharmaceutical composition and health functional food Can be usefully used.

In one aspect, the present invention provides a method of making a fermented beverage, Respectively; Filtering each of the extracts of Hwanggi, Bokbunja and Pork potato; Concentrating the filtrate of Hwanggi, Bokbunja and Pork potato extract respectively under reduced pressure; Lyophilizing each of the red ginseng, bokbunja, and porcine potato reduced-pressure concentrate; And bovine, bacterium, and porcine potato extracts comprising the step of mixing the lyophilisates of the respective hulls, brambles and pig potato extracts in a ratio of 3: 1: 2 (w / w).

In one embodiment, distilled water can be added in a ratio of 1: 5 to 1: 9 (w / v) to hull, brambly and porcine potatoes, and reflux extraction can be carried out at 70 to 100 ° C for 1 to 5 hours.

The present invention will be described in more detail with reference to the following examples. However, the following examples are only for the purpose of illustrating the present invention, and thus the present invention is not limited thereto.

Example 1. Preparation of hot-water extracts of Hwanggi, Bokbunja and Pork potatoes

700 ml of tertiary distilled water was added to 100 g of dried hwanggi, bokbunja, and porcine potatoes, and the process of reflux extraction at 90 ° C for 3 hours was repeated three times. Each of the extracted extracts was filtered, and then concentrated under reduced pressure at 40 ° C. The concentrated extracts were lyophilized for 7 days and used as extracts.

Example 2. Preparation of a composite of hot water extract of Hwanggi, Bokbunja and Pork potatoes

The hydrothermal extracts of each of the hwanggi, bokbunja and porcine potatoes prepared in Example 1 were mixed at a weight ratio of hwanggi: bokbunja: pig potato = 1.5: 0.5: 1.0 to prepare a composite.

Experimental Example 1. Inhibition of cell viability and glucose production in HepG2 hepatocytes induced by high glucose

In order to confirm the hepatocyte toxicity and the glucose production inhibition ability of the combination of the hwanggi, bokbunja and porcine potato hot water extracts prepared in the above examples, the cell viability and the glucose production inhibition ability of HepG2 hepatocytes induced by high glucose were confirmed. Specifically, HepG2 cells were seeded at a density of 5 × 10 ⁴ cells / well in a 96-well plate and cultured in DMEM-low glucose medium containing 10% FBS and 1% penicillin-streptomycin in an incubator under the conditions of 5% CO ² and 37 ° C. To stabilize for 24 hours. Subsequently, to remove diabetic condition, 100 μL of DMEM-high glucose-serum-free (DMEM-high-SF) medium was dispensed into each well, and the cells were stabilized for 24 hours in an incubator under the same conditions. Respectively. Thereafter, the hydrothermal extract compound prepared in Example 2 was treated at 100, 250, 500 and 1000 / / ml, respectively, and cultured. For reference, a group not treated with the extract was used as a control group. After 18 hours of culture, the medium was collected and the sugar content was measured using a blood glucose measurement kit (ASAN PHARM.CO., Ltd). To confirm cell viability, 50 μl of MTT solution (2 mg) was added to a 96-well plate from which the medium had been removed, reacted in a 5% CO ² incubator at 37 ° C. for 2 hours, MTT solution was removed, And the mixture was reacted at room temperature for 20 minutes. After dissolving the insoluble formazan crystals formed through the reaction, the absorbance was measured at 540 nm using a microplate reader (BIO-RAD 450, California, USA).

As a result, when the complex of hwanggi, bokbunja and porcini potato hot water extract of the present invention was treated, cytotoxicity was not observed even at a high concentration of 1000 ug / ml (Fig. 1). In addition, from the concentration of 100 ug / ml of the complex, saccharine production inhibitory effect was shown, and the highest saccharide formation inhibitory activity was shown at 1000 ug / ml (Fig. 2).

Experimental Example 2. Comparison of Cell Survival and Blood Glucose Reducing Effect According to Mixing Ratios of Hot Water Extract of Hwanggi, Bokbunja and Pork Potatoes

In order to compare the blood glucose lowering effect according to the mixing ratio of the mixture of the hwanggi, bokbunja, and porcine potato hot water extracts prepared in the above examples, the effect of reducing glucose in HepG2 hepatocytes induced by high glucose was confirmed. Specifically, HepG2 cells were seeded at a density of 5 × 10 ⁴ cells / well in a 96-well plate and cultured in DMEM-low glucose medium containing 10% FBS and 1% penicillin-streptomycin in an incubator under the conditions of 5% CO ² and 37 ° C. To stabilize for 24 hours. Subsequently, to remove diabetic condition, 100 μL of DMEM-high glucose-serum-free (DMEM-high-SF) medium was dispensed into each well, and the cells were stabilized for 24 hours in an incubator under the same conditions. Respectively. Thereafter, the hydrothermal extract compounds prepared in Example 2 were mixed at the mixing ratios shown in Table 1, and then treated at 100, 250, 500 and 1000 μg / ml, respectively, and cultured. For reference, a group not treated with the extract was used as a control group. After 18 hours of culture, the medium was collected and the sugar content was measured using a blood glucose measurement kit (ASAN PHARM.CO., Ltd). To confirm cell viability, 50 μl of MTT solution (2 mg) was added to a 96-well plate from which the medium had been removed, reacted in a 5% CO ² incubator at 37 ° C. for 2 hours, MTT solution was removed, And the mixture was reacted at room temperature for 20 minutes. After dissolving the insoluble formazan crystals formed through the reaction, the absorbance was measured at 540 nm using a microplate reader (BIO-RAD 450, California, USA).

Hwanggi Bokbunja Pig potato Remarks Type 1 One One One 3 mg / mL 333 l 333 l 333 l 1000 μl Type 2 1.25 1.25 0.5 3 mg / mL 417 μl 417 μl 167 μl 1000 μl Type 3 0.5 1.25 1.25 3 mg / mL 167 μl 417 μl 417 μl 1000 μl Type 4 1.25 0.5 1.25 3 mg / mL 417 μl 167 μl 417 μl 1000 μl Type 5 1.5 0.5 1.0 3 mg / mL 500 μl 167 l 333 l 1000 μl Type 6 1.0 1.5 0.5 3 mg / mL 333 l 500 μl 167 l 1000 μl Type 7 0.5 1.0 1.5 3 mg / mL 167 l 333 l 500 μl 1000 μl

As a result, hepatocyte showed the blood glucose lowering effect at all of the above mixing ratios. Particularly, at a concentration of 1000 ug / ml, the mixture ratio of Hwanggi, bokbunja and porcine potato hot water extract was 3: 1: 2 (w / (Figure 3). ≪ tb > < TABLE > In addition, cytotoxicity was not observed even at a high concentration of 1000 ug / ml in all of the types 1 to 7 (Fig. 4).

EXPERIMENTAL EXAMPLE 3 Expression of Related Protein in Sugar Production

In order to confirm the hepatocyte toxicity and the glucose production inhibition ability of the combination of the hwanggi, bokbunja, and porcine potato hot water extracts prepared in the above examples, the expression of sugar-related protein expression in high glucose-induced HepG2 hepatocytes was confirmed. Specifically, HepG2 cells were seeded at a concentration of 5 × 10 ⁶ cells / well in a 100 mm dish and incubated in DMEM-low glucose medium containing 10% FBS and 1% penicillin-streptomycin in an incubator under the conditions of 5% CO ² and 37 ° C. To stabilize for 24 hours. Then, 10 mL of DMEM-high glucose-serum-free (DMEM-high-SF) medium was dispensed into each dish, and the cells were stabilized for 24 hours in an incubator under the same conditions. High glucose Lt; / RTI > Then, the hot-water extract composite prepared in Example 2 was treated at a concentration of 100, 250 and 500 μg / ml, respectively. The positive control was treated with metformin and cultured for 24 hours each. Cells that were completely filled after incubation were lysed using lysis buffer containing 150 mM NaCl, 1% NP40, 0.5% Sodium Deoxycholate, 0.1% SDS and 50 mM Tris-Cl. Thereafter, the protein content of the lysate supernatant was measured with Bio-Rad Protein Assay (Bio-Rad, California, USA) using BSA as a standard, and the total protein content of each sample was adjusted. For subsequent electrophoresis, the samples were loaded onto 8-15% SDS-PAGE gels to a protein amount of 30 μg each, and transferred to nitrocellulose membranes. The membranes were blocked with 5% skim milk and incubated with a chemiluminescence detection system (Bio-Rad, USA) using antibodies against PEPCK, GSK3?, P-GSK3 ?, GLU4 and Actin (Cell Signaling Technologies, California, USA).

As a result, it was found that the complex of the present invention decreased the expression of PEPCK involved in glucose production induced by high glucose, and the expression of GSK-3β, which promotes accumulation of glycogen, was increased (Fig. 5). Through this, it was possible to suggest that glucose accumulation in the blood glucose is stored in the cells by accelerating the accumulation of glycogen, thereby lowering the blood glucose. On the other hand, metformin, a positive control, was found to increase the expression of GLU4, which is involved in the storage of glucose into cells.

Experimental Example 4. Antidiabetic effect in a diabetic induction animal model of a complex

4-1. Animal models and methods of administration

Db / db mice, an animal model in which type 2 diabetes was induced, were purchased from central laboratory animals and tested. The control group used the heterotypic m + / db. Eight groups per each group were divided into five groups as shown in Table 2 below. Samples were prepared by dissolving in water at a concentration of 10 mg / ml.

group Animal model Administered water Normal group m + / db - Diabetic induction group db / db - Compound administered group 1 db / db 50 mg / kg complex Compound administered group 2 db / db 100 mg / kg complex Positive control group db / db 200 mg / kg metformin

4-2. Check for fasting blood sugar

The blood glucose level of each experimental animal model group was measured once every two weeks, and the dietary supply was stopped the day before the measurement, and the fasting state was maintained for 12 hours and the blood glucose was measured. Blood was drawn from the tail of each mouse and measured using a glucose meter (Accu-Chek Performa).

As a result, it was found that the test group administered with the extract compound of the present invention had lower blood glucose level than the diabetic group, especially when the compound was administered at 100 mg / kg for 8 weeks, (Fig. 6).

4-3. Glycosylated hemoglobin

The glycated hemoglobin of each experimental animal model group was measured once in the tail as in Experimental Example 3-2, and was measured using a glycated hemoglobin analyzer (Infopia Co., Ltd. Easy A1c).

As a result, the glycated hemoglobin level of the test group administered with the extract compound of the present invention at a dose of 100 mg / kg was lower than that of the db / db group, which was induced in the diabetic group, and the degree of decrease was similar to that of the metformin group (FIG. 7).

4-4. OGTT (Oral glucose tolerance test)

In order to measure the OGTT of each experimental animal model group, the dietary intake of the animal model was stopped the day before the measurement and the fasting state was maintained for 12 hours. Thereafter, glucose was administered at a dose of 1 g / kg. Blood glucose was measured at 15, 30, 60, 90 and 120 minutes after glucose administration by the same method as Experimental Example 3-2.

As a result, the blood glucose level temporarily increased due to the glucose administration did not decrease at all after 2 hours in the diabetic induction group db / db, but in the group administered with the complex of the present invention, the blood glucose level was lower than that of the positive control group metformin (Fig. 8).

Claims (9)

A composition for preventing or treating diabetes mellitus, comprising a combination of extracts of hwanggi, bokbunja and pork potato as an active ingredient. The composition for prevention or treatment of diabetes according to claim 1, wherein the extract of hwanggi, bokbunja and porcini potatoes is a mixture of hwanggi, bokbunja and porcini potato extracts at a ratio of 3: 1: 2 (w / w). 3. The composition for preventing or treating diabetes according to claim 2, wherein the extract of hwanggi, bokbunja and porcini potato is a hot-water extract. A composition for extracting hwanggi, bokbunja and pork potato as an active ingredient. The food composition according to claim 4, wherein the hull, bramble and pig potato extract complexes are mixed with 3: 1: 2 (w / w) of haggis, brambly and porcine potato extract. The food composition according to claim 4, wherein the hwanggi, bokbunja, and porcini potato extracts are hot-water extracts. 1) adding distilled water to the hwanggi, bokbunja and pig potato respectively;
2) is refluxed;
3) filtering each of Hwanggi, Bokbunja and Pork potato extract;
4) concentrating the filtrate of Hwanggi, Bokbunja and Pork potato extract respectively under reduced pressure;
5) lyophilizing each of Hwanggi, Bokbunja and Porky potato depressurized concentrate; And
6) mixing the lyophilisates of the respective hwanggi, bokbunja and porcini potato extracts in a ratio of 3: 1: 2 (w / w). The process according to claim 7, wherein distilled water is added in a ratio of 1: 5 to 1: 9 (w / v) to the hulls, brambles and porcine potatoes in step 1). The process according to claim 7, wherein the reflux extraction in step 2) is carried out at 70 to 100 캜 for 1 to 5 hours.

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