KR100962292B1 - Pharmaceutical composition for preventing and treating diabetes comprising extract of Mulberry leaves-Phellinus linteus mycelia - Google Patents

Abstract

The present invention relates to a mulberry leaf mushroom fungus mycelium extract having a hypoglycemic function without side effects, a pharmaceutical composition for preventing or treating diabetes containing the extract as an active ingredient and a health food for preventing or treating diabetes containing the extract as an active ingredient will be.

The mulberry leaf situation mushroom mycelium hot water extract concentrate according to the present invention has the effect of suppressing the post-prandial blood sugar rise without side effects in diabetics or people with diabetes risk factors that require blood sugar control, added to foods that cause blood sugar rise It can be used as a raw material.

Mulberry Leaf Strain Mushroom Mycelia Extract, Diabetes, Pharmaceutical Composition, Health Food

Description

Pharmaceutical composition for preventing and treating diabetes comprising extract of Mulberry leaves-Phellinus linteus mycelia}

The present invention relates to a fermented mulberry leaf mushroom fungus mycelium extract having an effect on suppressing post-prandial blood sugar elevation, more specifically, a mulberry leaf mushroom fungus mycelium extract having a hypoglycemic function, preventing diabetes or containing the extract as an active ingredient The present invention relates to a therapeutic pharmaceutical composition and health food.

Diabetes mellitus is one of the most common chronic adult diseases. In Korea, diabetic patients are estimated to be at least 2.5 million, about 5% of the total population. In developed countries, diabetic patients are reported to 10% of the total population. In addition, it is expected that the number of diabetics will gradually increase as the lifestyle becomes westernized. In particular, due to the surge in the elderly population, interest in elderly diabetes is increasing, and about 10% of the elderly aged 65 or older may have diabetes.

Diabetes is a disease caused by a lack of hormones that regulate blood sugar (glucose in the blood) in our body or a disorder of glucose metabolism due to the lack of hormones that function on each organ of the body. Energy sources in the body, especially the use of sugar is not good enough to lack the necessary energy, while unused sugar is accumulated in the blood more than necessary to be discharged into the urine.

Blood sugar is glucose from the food we eat and is used as an energy source by acting as a kind of fuel in the human body. In normal people, when blood sugar rises due to foods consumed after a meal, insulin is secreted from the pancreas (interest) according to the degree, at which time the secreted insulin carries sugar (glucose) in the blood and uses it as energy. In the case of diabetic patients, the blood sugar level is normal, but in the case of diabetics, the blood sugar rises due to insufficient insulin secretion or inability to function properly. Exhausted and a large amount of glucose comes out with the water is discharged because of a lot of urine volume, energy loss and fatigue, and eating a lot of weight loss occurs, such as a variety of abnormalities.

Hyperglycemia is a condition in which blood sugar levels rise above normal, which is a big problem because it causes chronic complications as well as acute symptoms as described above. In other words, if hyperglycemia persists, the primary goal of diabetes treatment is to maintain high blood sugar levels by lowering high blood sugar levels because of the high possibility of complications such as atherosclerosis in larger blood vessels as well as microvascular complications such as blood vessels.

Diabetes is largely divided into type 1 diabetes and type 2 diabetes. Type 1 diabetes is insulin-dependent diabetes, which usually occurs before the age of 30, and is usually low in weight. Is most common in children aged 11-12 years, but rarer in Asians than in Westerners.

Since type 1 diabetes is caused by the destruction of insulin-secreting cells in the pancreas and the absolute lack of insulin, insulin injection is essential for the treatment of diabetes. Type 2 diabetes is insulin-independent diabetes, usually occurring after age 40. In many cases, obesity and symptoms appear slowly, so the symptoms may not be obvious. Unlike type 1 diabetes, the pancreatic function is normally close to normal in the early stages, so insulin is normally secreted, but insulin is functioning in the organs where the insulin works (muscles, liver, fat cells, etc.). It is rare to fall into acute ketoacidosis like type 1 diabetes because it is a diabetes caused by insufficient exercise, and insulin injection is not essential in terms of treatment.

Type 2 diabetes accounts for over 90% of all diabetic patients, and the incidence increases with age and obesity, and about half of the patients are obese over normal weight. Therefore, the treatment of type 2 diabetes is insufficient insulin, but the pancreatic insulin secretion ability remains, diet therapy and exercise therapy may be able to control blood sugar, if the blood glucose control is not possible by such a method (oral blood glucose) Taking blood sugar) or using insulin injections to control blood glucose.

Recently, the development of anti-diabetic agents has been actively developed, and several types of therapeutic agents have been developed. In the past, the use of oral hypoglycemic agents, such as fast-acting insulin or sulfonurea, has been the main treatment with simply lowering elevated blood sugar, and these therapies are difficult to normalize glucose metabolism and thus prevent complications.

Since the epidemiologic investigation of the relationship between postprandial hyperglycemia and cardiovascular disease has been published, the importance of postprandial hyperglycemia control has been emphasized in diabetics. Accordingly, as the importance of the first secretion of insulin secretion in the blood sugar control has been highlighted, drugs or foods for reducing the occurrence of hypoglycemia have been attracting attention.

Alpha-glucosidase, a type of digestive enzyme, is known to be involved in the conversion of carbohydrates into glucose, which is absorbed into the blood from the small intestine and directly involved in raising the concentration of sugar in the blood. Inhibiting the activity of this enzyme inhibits the absorption of glucose into the blood, so the mechanism of action that can prevent the rise of blood sugar after eating is well known fact. Alpha-glucosidase inhibitors, unlike other drugs, have the advantage of not causing hyperinsulinemia or hypoglycemia. Therefore, since it is an effective drug for controlling postprandial hyperglycemia, it is used as an important therapeutic agent for elderly diabetics or patients with impaired glucose tolerance, and because of its unique mechanism of action, an additional glycemic effect can be expected in combination therapy with other drugs.

Mulberry trees ( Morus) alba L. and other plants of the genus Morus ) are inhabited in Japan, Korea, and China, and their leaves are used as food for silkworms producing cocoons.

In Japan, fibroin protein solutions, gels and powders are processed from cocoons into skin creams, face wash milk, shampoos and bath products. Mulberry leaves are used to treat diabetes in Chinese medicine. The bark of the mulberry root is called baekbaekpi, and it is used as an anti-inflammatory, diuretic, cough medicine, antipyretic medicine, and the mulberry fruit is called audi and is used as a tonic or sedative.

In 1976, an agrochemist isolated 1-deoxynojirimycin from the mulberry root bark and named it moranoline in Japan. Later, it was found that 18 sugar-mimic alkaloids, including 1-deoxynojirimycin, were isolated from mulberry root bark and leaves and inhibited mammalian α-glucosidase.

Phellinus linteus ; Woody mud mushrooms are mushrooms belonging to the family of pine scaly mushrooms. The situation has been known as a valuable herbal medicine since ancient times and has been used for antiperspirant or gynecological diseases.

Situation mushroom fruiting hot water extract has been shown to have a slowing effect on cancers of the digestive system and to increase immune function by combination of chemotherapy after liver cancer and cancer resection surgery, and many studies have been conducted. Proved by

Conventional natural situation mushrooms are difficult to breed in the natural world and rarely grow native to mulberry trees, but the situation mushrooms or mycelium grown by artificial cultivation methods using various culture conditions are used, but the intake requirements and dietary methods are not easy. Is on the side. Situation mushroom rice is a cultivated mushroom mycelium mass that has been invaded and changed into the whole surface of rice and the inside of rice by cultivating the fungi mushrooms as a medium and then cultivating it using a grain of rice as a medium. It is a valid invention with a variety of pharmacological functions and is registered as a Korean patent (No. 02,049,82,049,83,0,049,84,0,250,49,0,250,50).

Until now, research and development of foods using mulberry leaves or situation mushrooms have been concentrated on the identification of manufacturing methods and beverages that have been traditionally taken from the private sector, and are limited to anticancer supplements and blood sugar control supplements.

The present invention was devised by the above requirements, the present invention is mixed with dried mulberry leaf powder and rice in a certain ratio, the situation mushroom hyphae (Phellinus linteus, wood mud mushroom) cultured purely after sterilization in autoclave for 1 hour It was inoculated in sterile state, dried mycelium purely cultivated for a certain period of time at a certain temperature, and then pulverized and then extracted with hot water for 2 to 4 hours. To provide.

In order to solve the above problems, the present invention provides a mulberry leaf situation mushroom mycelium extract having a hypoglycemic function without side effects.

The present invention also provides a pharmaceutical composition for preventing or treating diabetes containing the extract as an active ingredient.

In addition, the present invention provides a health food for preventing or treating diabetes containing the extract as an active ingredient.

The mulberry leaf situation mushroom mycelium hot water extract concentrate according to the present invention has the effect of suppressing the post-prandial blood sugar rise without side effects in diabetics or people with diabetes risk factors that require blood sugar control, added to foods that cause blood sugar rise It can be used as a raw material.

In order to achieve the object of the present invention, the present invention provides a mulberry leaf situation mushroom mycelium extract having a hypoglycemic function without side effects.

The mulberry leaf mushroom fungus mycelium can be obtained by mixing the dried mulberry leaf powder with rice and sterilized in a sterile state by inoculated in a sterile state after incubated in a high-pressure sterilizer in a sterile state, dried mycelium, and then pulverized . The mixing ratio of the dried mulberry leaf powder and rice is preferably 30:70 by weight.

The solvent for extracting the extract of the present invention may be a polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

Extracting mulberry leaf mushroom fungus mycelium of the present invention, after crushing and grinding the dried mulberry leaf mushroom fungus mycelium, about 5 to 25 times the weight (kg), preferably about 10 to 15 times water, 1 to 4 carbon atoms Lower alcohols of or about 1: 0.1 to 1:10, preferably 1: 0.2 to 1: 5 with a mixed solvent having a mixing ratio (kg / l) of 50 to 100 ℃, preferably 70 to 80 ℃ extraction Soluble extract according to water, lower alcohol, or a mixed solvent thereof is extracted, concentrated under reduced pressure and freeze-dried by an extraction method such as solvent extraction or hot water extraction for about 1 hour to 1 day, preferably 2 hours to 4 hours at a temperature. It can be obtained by. Preferably, the extract may be extracted for 2 to 4 hours with hot water.

In addition, the present invention provides a pharmaceutical composition for preventing or treating diabetes, comprising the extract of mulberry leaf mushroom mycelium of the present invention as an active ingredient.

The composition of the present invention comprises 0.1 to 80% by weight of the extract, preferably 1.0 to 50% by weight, based on the total weight of the composition.

Pharmaceutical dosage forms of the extracts of the invention may also be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other pharmaceutically active fractions.

The composition comprising the extract of the present invention may further comprise a suitable carrier, excipient or diluent commonly used in the manufacture of pharmaceutical compositions. Examples of carriers, excipients and diluents that can be included in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

Compositions comprising extracts according to the invention are each formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories or sterile injectable solutions according to conventional methods. Can be used. Specifically, it may be formulated using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. that are commonly used when formulated.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may comprise at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like can be mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Liquid preparations for oral use include suspensions, solvents, emulsions, and syrups, and may include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have.

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The amount of the extract of the present invention may vary depending on the age, sex, and weight of the patient, but generally in an amount of 0.01 to 500 mg / kg, preferably in an amount of 0.1 to 100 to 200 mg / kg, once or several times a day. It can be administered separately. In addition, the dosage of the extract may be increased or decreased depending on the route of administration, the type and severity of the disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The composition of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The composition of the present invention can be coated on the surface of grains such as rice, grains, and can be coated up to 0.1% to 10% by weight of the grain.

The mulberry leaf mushroom fungus mycelium extract itself of the present invention has little toxicity and side effects, so can be used with confidence even for long-term use for the purpose of prevention.

In addition, the present invention provides a health food containing the mulberry leaf situation mushroom mycelium extract of the present invention as an active ingredient.

When the mulberry leaf mushroom fungus mycelium extract of the present invention is used as a food additive, the mulberry leaf mushroom fungus mycelium extract may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method. The blending amount of the active ingredient can be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). In general, during the production of food or beverage mulberry leaf mushroom mushroom mycelium extract of the present invention is added in an amount of 0.1 to 15% by weight, preferably 0.2 to 10% by weight relative to the raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. Is sure.

There is no particular limitation on the kind of food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  One: Mulberry Leaf Strain Mushroom  Preparation of Extract

(1) establishment of mulberry leaf mushroom mycelium culture method

Situation mushrooms were transferred from slant to petri dish, incubated for 7 days in a 30 ° C incubator, shaken for 15 days in a 500 ml Erlenmeyer flask, and then mixed with soy flour and glucose in 10L air lift type glass bottles. It was inoculated in a medium prepared by sterilization and then incubated for 7 days and used as an inoculum.

After mixing the brown rice glutinous rice and mulberry in a polypropylene standard bag (200mm × 300mm) for sterilization of mushrooms and sterilizing it for 4 hours in an autoclave with a total weight of 340g, inoculate 5% of the prepared mushroom inoculum and mix it evenly to 30 ℃. The whole culture process was completed by incubation for 50 days in a controlled thermostat.

The mixing ratio of mulberry leaves and brown rice glutinous rice was cultured by adjusting the mulberry leaves to 5, 10, 15, 25, 50% of the total weight.When the ratio of mulberry leaves exceeds 25%, the air permeability worsens and the growth of the situation mushroom mycelium is about 50%. Decreased. Finally 30% was confirmed and this sample served as the test material for all experiments. Currently established culture methods can produce 2 tons of raw materials per month, so there is no problem in industrialization.

(2) Preparation of Mulberry Leaf Situation Mushroom Mycelia Extract

After drying the mulberry leaf mushroom mycelium cultured in the above (1), 1,000g of the dried mulberry leaf mushroom mushroom mycelium was pulverized into 100 ~ 200 mesh using a grinder, and hot water extracted from 3,000ml of distilled water for 2-4 hours. The extract was filtered and dried using a spray dryer to obtain a mulberry leaf status mushroom mycelium extract with a dry weight of 450g.

Example  2: of the present invention Mulberry Leaf Strain Mushroom  Hypoglycemic Effects of Extracts

α-glucosidase is an enzyme that breaks down oligosaccharides, trisaccharides, and disaccharides into monosaccharides in the small intestine.Acabose and deoxynojirimycin, which are inhibitors of α-glucosidase, are antidiabetic agents and are used to treat α-glucose on the ciliated border of the small intestine. Inactivates the cedase and functions to reduce or delay the digestion of carbohydrates.

Acarbose is an enzyme that digests carbohydrates and is used as a therapeutic agent for diabetes because it is an α-glucosidase inhibitor that reversibly decreases the function of α-amylase. However, because it inhibits the breakdown of polysaccharides, polysaccharides remain in the intestines, and the digestion of bacteria causes side effects such as flatulence and diarrhea.

1. Materials and Methods

Animal breeding and diabetes induction

(1) Animal species (family): S. D. rats (Sam taco)

(2) Breeding environment

ㅇ Breeding environment

The environment of this animal is 22.2 ± 0.9 ℃, relative humidity 52.2 ± 0.9%, ventilation frequency 13-18 times / hr, lighting time 12 hours (7 am~7pm), illuminance 200 ~ 300 Lux. As a condition, commercial feed and drinking water were fed to isolate and reared during the purification and study period.

ㅇ Breeding box

During the purifying and study period, three rats were placed in a polycarbonate breeding box (400 × 250 × 180 mm ³ ) and placed on large padded rice litter.

ㅇ Feed and negative

Feed fed freely the solid animal feed (Samyang Yuji Feed Co., Ltd.) for experimental animals.

(3) Grouping

ㅇ The experimental groups were divided into normal control group, diabetic control group, acarbose (acarbose) group, and mulberry leaf mushroom mycelium extract group.

(4) Diabetes induction and confirmation

Diabetes was induced by streptozotocin (STZ) injection of 50 mg / kg body weight through the tail vein. The control group was injected with the same amount of citric acid buffer. Diabetes induction was checked 48 hours later, blood was collected from the tail vein, and blood glucose was measured using a blood glucose meter.

Dosage and Method

Acarbose was orally administered before meals at 50 mg / kg body weight.

Mulberry leaf mushroom fungus mycelium extract was administered orally at 3000mg / kg body weight.

Blood glucose measurement over time

After 1 week of diabetes-induced fasting for 12 hours, and fasting blood glucose levels were checked. In the control group, saline was administered, and acarbose and mulberry leaf mushroom mycelium extracts were orally administered to the experimental groups, respectively, and fed with feed and water for 1 hour. Thereafter, blood glucose was measured for 2 hours at 30 minute intervals and for 3 hours (5 hours total) at 1 hour intervals. Blood glucose measurements were measured by collecting blood from the tail vein using ACCU-CHEK Active (Roche Diagnostics GmbH, Germany).

For enteric gas assessment CT  ( Computer tomography Shooting

Computer (Tomography) imaging was performed at Chungnam National University Animal Hospital for evaluation of intestinal gas 30 minutes after drug and mulberry leaf fungus mycelium extract administration. Respiratory anesthesia was performed with isoflurane, and the normal control group, the diabetic control group, the acarbose group, and the mulberry leaf fungus mycelium extract group were placed in the upper right, upper left, lower right and lower left, respectively. Photographs were taken at 2 mm intervals from the diaphragm as a starting point. A total of 10 cuts corresponding to 2 cm under the stomach were compared and analyzed. The average of the total cross-sectional area divided by the average of the area occupied by the gas was shown graphically.

Biochemical analysis

Anesthesia was induced by intraperitoneal injection of 16.5% urethane (1ml / 100g), followed by laparotomy and blood collection from the abdominal aorta. Blood was left at room temperature for 30 minutes and then serum was separated by centrifugation at 4 ° C and 7000 rpm for 15 minutes. Glucose, Blood Urea Nitrogen, Cholesterol, Albumin, Total Protein, Triglyceride, Alkaline phosphate, As Aspartate Transaminase, Alanine Transaminase levels were measured using Vettest 8008 (IDEXX-GmbH, Woerrstadt, Germany).

group processing  And morphological staining

After anesthesia by injection intraperitoneally, blood was collected from the abdominal vein and perfusion was performed. PBS (phosphate-buffered saline) was perfused for 5 minutes to wash the kidneys, and then fixed solution was perfused for 10 minutes for immunostaining after PBS washing. PLP (Periodate Lysin Paraformaldehyde), 2% paraformaldehyde, was used as a fixative. The fixed kidneys were stored on PLP after triming for immunostaining. At the end of tissue collection of each test animal, the test tool was perfused with PBS. Post-embedding immunocytochemistry requires the preparation of waxes. Overnight PEG and 55.5 g of acetyl alcohol were mixed and dissolved at 60 ° C. The tissue to be waxed was placed in 70% alcohol overnight. Soaked in 80%, 90%, 95%, 100% alcohol. Alcohol: Wax 1: 1 and then immersed in Pure Wax I and II. Glycerol was applied to the mold and the wax was poured. The tissues were quickly moved one by one. The hardened, hardened tissue was stored cold and then trimmed in a trapezoid. 5 micrometers cut with a microtome. After distilled water was placed on the slide, the cut tissue was raised, the slide was raised, the water was removed and placed on a hot plate overnight. The wax was removed on the slide, and the slide was dehydrated and then stained with Hematoxyin Eosin and Masson trichrome.

2. Results

end. Biochemical test

Serum chemistry resulted in glucose, blood urea nitrogen (BUN), cholesterol (CHOL), albumin (ALB), total protein (TP), triglycerides (TRIG), alkaline phosphatase (ALKP) and aspartate amino. Transferase (AST), alkali aminotransferase (ALT) levels were measured using Vettest 8008 (IDEXX-GmbH, Woerrstadt, Germany) (Table 1).

Table 1. Biochemical Properties of Serum

^* P <0.05 vs normal rats. Data are expressed as mean ± standard deviation.

Diabetic: Streptozotocin-induced Diabetic Rats

Mulberry leaf: Oral Administration of Mulberry Leaf Situation Mushroom Mycelia Extracts in Diabetic Rats

Acarbose: Oral Administration of Acarbose in Diabetic Rats

The levels of ALB and TP were increased in the mulberry leaf fungus mycelium extract treated group and the acarbose treated group compared to the diabetic control group, and the TRIG, ALKP, AST, and ALT levels were decreased in the mulberry leaf fungus mycelium treated group and the acarbose treated group compared to the diabetic control group.

BUN and CHOL levels were decreased in the mulberry leaf fungus mycelium extract group compared to the diabetic control group, but increased in the acarbose group.

I. Morphological examination

Hematoxylin-Eosin staining and Masson Trichrome staining for morphological examination showed no structural changes due to diabetes in glomeruli and extra glomerular matrix, especially glomerulosclerosis in Masson trichrome staining (Fig. 1). .

All. Blood glucose level change over time

The results of comparing and measuring changes in blood glucose levels at each time are shown in FIG. 2. One hour after the drug and mulberry leaf mushroom mycelium extract administration, the diabetic control group and mulberry leaf mushroom mycelium extract group increased to 527.7 mg / dL and 553.0 mg / dL, respectively, and the acarbose group was 461.4 mg / dL. The increase was small compared with that.

After 2 hours of administration, the diabetic control group and mulberry leaf fungus mycelium extract-treated group showed 507.0 mg / dL and 501.2 mg / dL of blood sugar, respectively, and the acarbose-treated group remained similar to 456.0 mg / dL. Five hours after the administration of mulberry leaf mushroom mycelium extract group showed a decrease in blood glucose to 325.0 mg / dL, which showed a similar decrease rate to the akabose group reduced to 267.2 mg / dL. In comparison, the diabetic control group showed a small decrease of 363.0 mg / dL.

la. Intestinal gas comparison evaluation

A total of 10 cuts corresponding to 2 cm under the stomach were analyzed and analyzed (FIG. 3), and the graph of the numerical value obtained by dividing the average of the total cross-sectional area by the average of the area occupied by the gas (FIG. 4).

In the acarbose-treated group, the intestinal gas increased by 160% compared to the diabetic control group, and the mulberry leaf situation mushroom mycelium extract-administered group reduced the intestinal gas by 25% compared to the diabetic control group.

Therefore, the mulberry leaf mushroom mycelium extract produces less intestinal gas than acarbose, it can be seen that fewer side effects than acarbose.

Example  3: of the present invention Mulberry Leaf Strain Mushroom  Α- of extract Glucosidase  Inhibition assay

100 mg of rat intestinal acetone powder was added to 3 ml of 0.9% NaCl solution and sonicated in 12 ice-cooled water baths for 30 seconds, followed by centrifugation at 10,000 g at 4 ° C. for 30 minutes. Only the supernatant was used directly for analysis or stored at -20 ℃. 50 μl of sample solution was added to 100 μl of rat α-glucosidase solution, and then allowed to stand at 37 ° C. for 10 minutes. 50 μl of 5 mM pNPG (p-nitrophenyl beta-glucoside) solution was added, followed by reaction at 37 ° C. for 15 minutes, and absorbance was measured using an ELISA reader at 405 nm to analyze rat α-glucosidase inhibitory activity (FIG. 5).

As can be seen in Figure 5, as the concentration of the mulberry leaf mushroom mushroom mycelium extract of the present invention was found to increase the rat α-glucosidase inhibitory effect.

Example  4: of the present invention Mulberry Leaf Strain Mushroom  Α- of extract Amylase  Inhibition assay

200 μl of the sample solution was added to 300 μl of 1 U porcine pancreatic α-amylase solution dissolved in 0.02 M sodium phosphate buffer (pH 6.9, containing 0.006 M sodium chloride) and incubated at 25 ° C. for 10 minutes. To this solution was added 500 µl of a 1% starch solution incubated at 25 ° C for 10 minutes in advance and allowed to react at 25 ° C for 10 minutes. The reaction was stopped by adding 1 ml of 1% DNS solution dissolved in 30% Rochelle salt, and then treated in a boiling water bath for 5 minutes, cooled to room temperature, and 10 ml of distilled water was added thereto. The absorbance of the reaction solution of the sugar and DNS solution decomposed from the substrate by α-amylase was measured using an ELISA reader at 540 nm, and a solvent in which the sample was dissolved instead of the sample was used as a control (FIG. 6).

As can be seen in Figure 6, the mulberry leaf situation mushroom mycelium extract administration group of the present invention was found to reduce the α-amylase inhibitory effect. In other words, α-amylase activity increases, it was found that the digestion is good.

1 shows the results of Hematoxylin-Eosin staining (top panel) and Masson Trichrome staining (bottom panel) of rats treated with various treatments.

A: normal control group, B: diabetes control group, C: oral administration group of acarbose to diabetic rats, D: oral administration group of mulberry leaf mushroom fungus mycelium extract to diabetic rats.

Figure 2 is a graph showing the effect of mulberry leaf mushroom mycelium extract and acarbose on blood glucose levels in streptozotocin-induced diabetes.

Normal: Normal control

Diabetic: Streptozotocin-induced Diabetic Rats

Mulberry: Oral administration group of mulberry leaf fungus mycelium extract in diabetic rats

Acarbose: Oral administration of acarbose to diabetic rats

3 is a result showing a comparison of intestinal gas ratio from computed tomography. A: 12 mm, B: 16 mm, C: 20 mm, D: 22 mm, E: 26 mm, F: 30 mm from the diaphragm as a starting point.

NCTL: normal control, DMCTL: diabetic control, ACAR: acarbose, MULB: mulberry leaf mushroom mycelium extract

4 is a graph showing the measurement of intestinal gas levels in the experimental group.

5 is a result of α-glucosidase inhibition of the mulberry leaf mushroom fungus mycelium extract.

Figure 6 shows the α-amylase inhibition results of mulberry leaf mushroom mycelium extract.

Claims (7)

After mixing the dried mulberry leaf powder and rice sterilized in autoclave, inoculated purely cultured mushroom mycelium in aseptic state, dried mycelium purely cultured, and then dried mulberry leaf mushrooms mycelium obtained by crushing blood sugar drop Mulberry leaf fungus mycelium extract having a function. delete The extract according to claim 1, wherein the solvent is a polar solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The extract according to claim 1, wherein the extract is hydrothermally extracted. A pharmaceutical composition for preventing or treating diabetes, comprising the extract of the mulberry leaf mushroom fungus mycelium of any one of claims 1, 3 to 4. The pharmaceutical composition according to claim 5, wherein the mulberry leaf mushroom fungus mycelium extract is contained in an amount of 0.1 to 80% by weight based on the total weight of the composition. A health food for preventing or treating diabetes, comprising the extract of mulberry leaf mushroom fungus mycelium of any one of claims 1, 3 to 4.

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