KR20100111088A - Composition for preventing and treating diabetes mellitus or diabetic complications comprising extract of herbal combination - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing complex herb extract for preventing and treating diabetes and complication is provided to prevent Langerhans islet contraction. CONSTITUTION: A pharmaceutical composition for preventing or treating type 1 diabetes or complication contains complex herb extract. The complex herb extract contains Rosa davurica Pall berries, Acanthopanax senticosus berries, acanthopanax senticosus bark, ginseng, alder rhytidome and Kalopanax ricinifolium rhytidome in a ratio of 1-10: 1-5: 1-5: 1-5: 1-10: 1-10. A food composition for preventing or relieving type 1 diabetes or complication contains the complex herb extract as an active ingredient.

Description

Composition for preventing and treating diabetes mellitus or diabetic complications comprising extract of herbal combination}

The present invention relates to a composition for the prevention or treatment of diabetes mellitus or its complications, which comprises extracts of bio-fruit, thorny oak fruit, thorny oak bark, ginseng, alder bark and oak bark as active ingredients, The present invention relates to a pharmaceutical composition for preventing or treating type 1 diabetes or its complications, and a food composition for preventing or alleviating prickly bark fruit, prickly bark bark, ginseng, alder bark and oak bark as active ingredients.

Diabetes threatens the health of about 15 million people worldwide and is expected to double in 20 years. In 2003, a survey of diabetics who visited a medical institution targeting 20-79 year-olds nationwide found that the prevalence of diabetes among Korean adults was 7.7%, with an additional 10% of new patients each year. Including patients with impaired fasting glucose and impaired glucose tolerance in all stages of diabetes, the occurrence of diabetes is expected to increase further.

Diabetes is largely divided into type 1 diabetes and type 2 diabetes. Type 1 diabetes is an insulin-dependent type that is absolutely deficient in insulin due to the destruction of pancreatic islets by T cell mediated autoimmunity, and type 2 diabetes is obesity. Insufficient exercise, stress, etc., combined with genetic predisposition increases insulin resistance or decreases the insulin sensitivity of the cell.

In diabetes, hyperglycemia (abnormally high blood sugar levels), diabetes (excessive glucose excretion in the urine) and glycogen lowering in the liver cause various metabolic disorders. These metabolic disorders lead to degenerative complications. For example, diabetic retinopathy, diabetic neurosis, nephropathy, atherosclerosis, myocardial disorders, skin disorders, diabetic foot lesions and other peripheral vascular diseases. In general, it is generally understood by improving hyperglycemia of diabetic patients to delay or prevent such degenerative complications. In addition, severe hyperglycemia may cause hyperosmotic biketone coma, so lowering blood glucose levels may prevent hyperosmotic biketone coma caused by hyperglycemia.

Treatment and management of diabetes is a global concern, and successful treatments are still being studied. Langerhans Island is an endocrine gland tissue scattered in the pancreas of insulin-secreting vertebrates discovered by German pathologist Langerhans in 1869. Beta-cells in the island of Langerhans are known to secrete insulin, which can lead to insulin deficiency. Recently, a method of directly administering the animal or human Langerhans islet cells to a patient has been attempted, but it is still in an experimental stage, and there are many problems to be solved, such as tissue rejection and supply amount.

Until now, insulin and oral hypoglycemic agents have been used as treatments.If you have type 1 diabetes, you will have clinical symptoms such as hyperglycemia, Daum, polyps, polyuria, and weight loss. Insulin therapy is known to be essential. The goal of treatment for type 1 diabetes is to maintain normal blood sugar to prevent acute complications. Insulin therapy is the ultimate treatment of type 1 diabetes, but type 1 diabetes is a long-term step, and there is a risk of hassle or complications that burden the patient. Therefore, there is a need for the development of a natural substance diabetic agent that can be fundamentally cured while minimizing the risk of side effects by inhibiting blood sugar and restoring Langerhans islet cells.

The inventors of the present invention while studying a composition composed of natural ingredients effective in diabetes and its complications, the composition consisting of extracts of bark fruit, spiny bark fruit, spiny bark bark, ginseng, alder bark and oak bark suppresses diabetes And the therapeutic activity was found and the present invention was completed.

Accordingly, an object of the present invention is 1 to 10: 1 to 5: 1 to 5: 1 to 5: 1 to 5: 1 to 10: 1 to 5: 1 to 5: 1 to 5: 1 to 10 on the basis of dry weight of each wild medicinal herb, barberry, bark, bark, bark, oak bark It is to provide a pharmaceutical composition for the prevention or treatment of type 1 diabetes mellitus or complications thereof as an active ingredient extract extracted by mixing at a ratio of 1 to 10.

Still another object of the present invention is 1 to 10: 1 to 5: 1 to 5: 1 to 5: 1 to 5: on the basis of dry weight of the wild bark fruit, spiny bark fruit, spiny bark bark, ginseng, alder bark and oak bark It is to provide a food composition for preventing or alleviating complications of type 1 diabetes or its complications, wherein the extract extracted by mixing at a ratio of 10: 1 to 10 as an active ingredient.

In order to solve the above problems, the present invention is 1 to 10: 1 to 5: 1 to 5: 1 on the basis of the dry weight of fresh herbs, thorny oak bark, thorny oak bark, ginseng, alder bark and oak bark To 5: 1 to 10: provides a pharmaceutical composition for preventing or treating type 1 diabetes mellitus or complications thereof by using the extract extracted by mixing in a ratio of 1 to 10 as an active ingredient.

In order to achieve the other object of the present invention, the present invention is 1-10: 1 to 5: 1 to 5: 1 on the basis of the dry weight of wild berry fruit, spiny bark fruit, spiny bark bark, ginseng, alder bark and oak bark 5 to 1 provides a food composition for preventing or alleviating complications of type 1 diabetes or the complications thereof, comprising the extract extracted by mixing in a ratio of 1 to 10: 1 to 10 as an active ingredient.

Hereinafter, the present invention will be described in detail.

The bio-fruit of the present invention refers to the fruit of Rosa davurica Pall. The red-green shrub is a perennial plant belonging to the family Rosacea, which is a deciduous broad-leaved shrub distributed in Korea, Japan, Manchuria, and Siberia. Since ancient times, it has been used for edible food, and it is known that the medicinal effects of roots and fruits include dry stomach and bleeding, and have been used for the treatment of indigestion, anticipation, stomach pain, and menstrual relief. In particular, the fruit contains abundant vitamin C, which is used as a tonic drink in the Chinese people.

Prickly Pear Fruit and Bark are the fruits and bark of Acanthopanax senticosus. Acanthopanax senticosus (Acanthopanax senticosus) is a deciduous broad-leaved shrub belonging to the genus Araliaceae. 효능), Ikgi Anshin (益氣 安神), is known to have the effect of active blood circulation (活血 通 絡) has been widely used in the treatment of pediatric musculoskeletal weakness, each myeloma, rheumatoid arthritis.

Ginseng has long been recognized for its medicinal effects based on thousands of years of experience in oriental medicine.In ancient books such as the New Nongbyeon Herbal Medicine and Boncho River, the ginseng mainly stabilizes and stabilizes the spirit. It has been known to stop vigilance, brighten the eyes, make the head wise and prolong the lifespan, and in modern times it has been found to restore liver function, anti-cancer immunity, anti-stress, fatigue recovery and carbohydrates. , Lipids, protein metabolism, etc., especially effective in diabetic hypoglycemic effect and improvement of general subjective symptoms, ginseng hypoglycemic effect is very effective, such as synthetic diabetes drugs to reduce the risk of excessive diabetes Because it is not present, it can be used as an adjuvant therapy while taking with diabetes It is known that this is.

Alder bark refers to the bark of Alnus japonica Steud. Alder is a deciduous algae tree attached to the birch family, called Yuri-mok, or Red-yang, and tea in China. Alder's bark contains 5-10% tannin, taraxolol and betulinic acid. Fruits are astringent diarrhea 멎 diarrhea, used for diarrhea, gastrointestinal diseases, the skin is used as postpartum blood 약 pill, stomach disease, and used for eye irritation, rheumatism, throat irritation, sunscreen or nausea. Alder has a taste and bitterness, and is cool in nature, reducing heat and poisoning. In particular, it is very effective in dissolving alcohol poison and has excellent therapeutic effect on various liver diseases such as hepatitis, cirrhosis and fatty liver.

The oak bark refers to the bark of the oak tree (Kalopanax pictus). The oak tree is a deciduous tree belonging to the family Elmaceae, with over 80 genera and 900 species distributed worldwide, and 8 genera and 20 species are native to Korea. The main distributions are Japan, Sakhalin, Okinawa and China. In Korea, it grows in the sunny area at the foot of 100 ~ 1,800m altitude of Jeollabuk-do, Gangwon-do, etc., and the area around 400-500m is the central zone. Leaves are regenerated, palm-shaped, split into 5-9 branches, heart-shaped, lobe is ovate with sharp tip and sharp sawtooth. Yin-Tak has been known for its bark in the oriental medicine, and its roots are known as hawthorn skin, and hawthorn root is called hawthorn root. It has been widely used as expectorant, analgesic, tonic and neuralgia. Moreover, recently immunological and antioxidant activities have been reported. Thawing blood is known to improve symptoms such as back and knee pain, eczema and skin diseases.

The extracts of the barberry, thorny bark, ginseng, ginseng, alder bark and oak bark of the present invention are extracted from the mixture of bark, barberry, barberry, ginseng, alder bark and oak bark Says liquid. The bark of the present invention, thorny oak fruit, thorny oak bark, ginseng, duck or radish bark and oak bark of 1 to 10: 1 to 5: 1 to 5: 1 to 5: 1 to 10: 1 Extract extracted by mixing at a ratio of 10 (hereinafter referred to as a mixed extract) can be prepared by a solvent extraction method known in the art. Examples of the extraction solvent include any one selected from alcohols having 1 to 6 carbon atoms such as water, ethanol and methanol, organic solvents such as acetone, ethyl acetate, n-nucleic acid, diethyl ether, acetone, and benzene, or a mixture thereof. Solvents may be used. Preferably, the extraction may be performed using water or an alcohol having 1 to 6 carbon atoms. When the extract is prepared by the solvent extraction method, hot water extraction, ultrasonic extraction and reflux extraction may be used. Most preferably, the mixed extract of the present invention may be prepared by a hydrothermal extraction method using water as a solvent. The ratio of fresh barberry, thorny oak bark, thorny oak bark, ginseng, alder bark and oak bark and water at the time of hot water extraction is not particularly limited, but water may be added 20 to 200 times (by weight). In order to increase the extraction efficiency, water may be added at 50 times to 100 times (by weight). The temperature during extraction is preferably performed at room temperature under atmospheric pressure and the extraction time depends on the extraction temperature, but is extracted for 5 hours to 15 hours, preferably 8 hours to 12 hours. In addition, the extraction efficiency can be further increased when stirring with a shaker (shaker) during extraction. In addition, in order to increase the extraction efficiency can be used to crush the bark bark, thorny oak fruit, thorny oak bark, ginseng, alder bark and oak bark with a grinder.

The ratios of bioscissus fruit, spiny bark fruit, spiny bark bark, ginseng, alder bark and oak bark used in the present invention are 1 to 10: 1 to 5: 1 to 5: 1 to 5: 1 to 10: dry weight basis. 1 to 10 are preferred, and most preferably 5: 3.5: 3.5: 1: 7: 7. At this time, in the case of using undried fruit, bark, etc., the weight ratio is calculated in consideration of the included moisture content.

In one embodiment of the present invention, 50g of fresh fruit, 35g of thorny oak fruit, 35g of thorny oak bark, 10g of ginseng bark, 70g of alder bark and 70g of oak bark in 20L of water for 5 hours at 80 ℃, 5 hours at 100 ℃ Extraction and lyophilization gave a mixed extract (see Example 1).

The mixed extract prepared by the ratio of the present invention described above shows an effect in the treatment of diabetes mellitus, and useful in the treatment of type 1 diabetes among diabetes. Thus, the present invention provides a pharmaceutical composition for treating type 1 diabetes containing the mixed extract prepared by the above ratio as an active ingredient.

The mixed extract of the present invention has the effect of preventing or treating type 1 diabetes. Pharmacological activity of the recovery of Langerhans islet cells in the pancreas with blood glucose lowering effect on the first symptoms of type 1 diabetes when the pharmaceutical composition containing the mixed extract of the present invention is applied to the treatment of type 1 diabetes The effect can be obtained.

The effect of preventing or treating the type 1 diabetes mellitus or complications caused by the mixed extract according to the present invention was confirmed by the present inventors through animal experiments.

That is, in one embodiment of the present invention, whether the mixed extract has anti-diabetic activity against rats of type 1 diabetic animal model induced by intraperitoneal administration of Streptozotocin (STZ, 65 mg / kg) through blood glucose change observation Confirmed.

As a result, all the diabetic group had significantly higher blood glucose level than the normal group. However, by the sixth week of the experiment, three of the six groups of the present invention with the combined extract of the present invention showed a blood glucose level slightly above normal blood glucose levels. As a result, the mixed extract of the present invention was confirmed to have excellent efficacy in reducing blood sugar (see Example 3).

In another embodiment of the present invention, it was confirmed by measuring histopathological changes whether the mixed extract has antidiabetic activity in a rat of type 1 diabetic animal model. Rats that induce type 1 diabetes were fed feed containing the fertilizer and the mixed extract of the present invention, and reared for 6 weeks, dissected, and the pancreas were extracted to observe the condition of the island of Langerhans.

Experimental results showed that the endocrine gland Langerhans is atrophy in all groups that induced diabetes with STZ, unlike the exocrine gland, which is not significantly different from the normal group. However, it was confirmed that the atrophy of the Langerhans island was recovered in the diabetic group fed the feed containing the mixed extract of the present invention (see Example 4-2).

In addition, pancreatic tissue samples were prepared and immunohistochemically tested using insulin antibodies to determine the level of insulin secretion in the beta cells of Langerhans islet. As a result, it was confirmed that the proportion of insulin-positive cells was higher in the diabetic group fed the feed containing the mixed extract of the present invention than the control group (see Example 4-3). As a result of confirming that the mixed extract of the present invention restores insulin secretion ability of type 1 diabetes, the mixed extract of the present invention proved to be excellent in type 1 diabetes.

The disease to which the composition according to the present invention can be applied may be applied to, but not limited to, diabetes and related complications, diseases caused by disruption or degradation of Langerhans islet cells, and diseases caused by an increase in blood glucose concentration. Specifically, but not limited to, type 1 diabetes, diabetic retinopathy, diabetic neurosis, kidney disease, atherosclerosis, myocardial disorders, skin disorders, diabetic foot lesions and other peripheral vascular diseases.

The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable suitable carrier, excipient and diluent, and pharmaceutically acceptable liquids, powders, granules, tablets, capsules, suspensions, emulsions, syrups, liquid formulations Oral formulations such as aerosols, and may be used in the form of external preparations, such as coating ointment.

Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like. In addition, the carrier for parenteral administration may include water, dropping oil, saline, aqueous glucose and glycols, and may further include stabilizers and preservatives. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.

The dosage of the pharmaceutical composition of the present invention depends on the type and severity of the disease, the type and content of the active ingredient and other ingredients included in the composition, the absorption of the active ingredient, the rate of release, the age and weight of the patient, sex and condition, etc. It may be appropriately selected, and in general, the adult may be administered once to several times a day at a dose of 1 mg / kg to 100 mg / kg, and preferably at a dose of 3 mg / kg to 10 mg / kg.

The present invention provides a food composition for preventing or alleviating complications of type 1 diabetes mellitus or a combination thereof as an active ingredient.

The food composition according to the present invention may be prepared in the form of a composition by mixing with a known active ingredient known to have a preventive and alleviating effect of type 1 diabetes or its complications, and may include a functional food and a nutritional supplement. And all forms, such as health food and food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art. For example, as a health food can be prepared in the form of tea, juice and drink containing the mixed extract of the present invention, it can be ingested by granulating, encapsulating or powdering a food composition comprising the mixed extract as an active ingredient. .

The preferred content of the mixed extract of the present invention may include about 0.1 to 50% by weight relative to the total weight of the food. Preferably intake of 0.5mg / kg / day ~ 10mg / kg / day in the amount may be effective to prevent and improve diabetes.

Therefore, the composition for the prevention or treatment of diabetes mellitus or its complications containing the extracts of the bark fruit of the present invention, thorny oak bark fruit, thorny oak bark, ginseng, alder bark and oak bark as an active ingredient, or Effective for treatment In particular, it has excellent efficacy in preventing or treating type 1 diabetes mellitus, chronic hyperglycemia, atherosclerosis, microangiopathy, kidney disease, heart disease and diabetic retinopathy. In addition, the food composition comprising the extract of the bark fruit, thorn oak bark fruit, thorn oak bark bark, ginseng, alder bark and oak bark of the present invention has an excellent effect on the prevention and alleviation of type 1 diabetes or its complications It can be used to manufacture health foods for patients, which is highly industrially applicable.

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 1 >

Preparation of Mixed Extracts of Fresh Berries, Prickly Pear Bark, Prickly Pear Bark, Ginseng, Alder Bark and Oak Bark

After drying 50g of dried wild berries, 35g of thorny oak bark, 35g of thorny oak bark, ginseng 10g, 70g of alder bark and 70g of oak bark, put into 20L of water and extract it for 5 hours at 80 ℃ and 5 hours at 100 ℃. The extract was lyophilized to prepare a mixed extract (hereinafter referred to as a mixed extract) of bark bark fruit, bark oak fruit, bark oak bark, ginseng, alder bark and oak bark.

<Example 2>

Manufacture of Type 1 Diabetic Animal Model

 The experimental animals purchased Spraque-Dawley male rats (weight: 190-220g) 7 weeks of age from OrientBio Co., Ltd., and fed the feed according to AIN-76G for a week and the new environment (temperature: 24 ± 5, humidity). : 50 ± 10, Contrast: 12 hours light / dark cycle).

 After 1 week of acclimatization, the rats weighed 240-290 g were fasted for 16 hours, and then Streptozotocin (STZ, 65 mg / kg) (Sigma Chemical Co., St Louis, Mo, USA) was dissolved in 10 mM pH 4.5 Sodium citrate buffer. After 3 days of intraperitoneal administration, fasting blood glucose was measured and a subject of 280 mg / dl or more was used as a diabetic group. An equal amount of sodium citrate buffer was administered to the control group.

<Example 3>

Physiological and Blood Glucose Measurements of Mixed Extracts of Fresh Berries, Prickly Pear, Prickly Pear Bark, Ginseng, Alder Bark and Oak Bark in Type 1 Diabetic Animal Model

<3-1> Administration of Mixed Extracts of Fresh Berries, Prickly Pear Bark, Prickly Bark Bark, Ginseng, Alder Bark and Oak Bark in Type 1 Diabetic Animal Model

 After administration of STZ and Buffer, the rats were divided into two groups, diabetic and normal control group, and they were divided into the following five experimental groups. Group A: Normal rats + Normal control diets; Group B: Normal rats + 0.01% extract diets; Group C: DM rats + Normal control diets; Group D: DM rats + 0.01% extract diets; Group E: DM rats + 0.1% extract diets

Feed and water were daily fresh and freely consumed.

Rat group used in the experiment Group Group classification Animal No. Group a Normal rats + normal control diets 6 Group b Normal rats + 0.01% extract diets 6 Group c DM rats + Normal control diets 6 Group d DM rats + 0.01% extract diets 6 Group e DM rats + 0.1% extract diets 6

<3-2> Physiological Change Observation of Mixed Extract Administration

In STZ-induced diabetic rats, carbohydrate and fat metabolism are not smooth and protein catabolism is increased, resulting in weight loss. Therefore, weight loss is one way to determine if diabetes is caused by STZ. Body weights were measured using electronic scales in the same order starting at week 0 at the time of grouping at 11 am on Monday morning for 6 weeks. The dietary intake was measured by the difference between the amount of feed per cage and the remaining amount at a given time every day to determine the total intake during the experiment. The amount of drinking water was calculated as the difference between the supply and the remaining amount during the 24-hour intake from Tuesday 11 to Wednesday 11 every week.

As a result, as shown in Figure 1, the normal rat body weight increased steadily during the experimental period (A, B) in the group fed the feed containing 0.01% extract and extract (156%, 6 weeks after 6 weeks, respectively). 151% increase. On the other hand, in the diabetic C, D, and E groups, the body weight was about 83% lower than that of the normal rats at 0 weeks, and increased or decreased slightly within 20 g for 6 weeks. At the end of the experiment, the weight of diabetic rats was about 50% of that of normal rats, and there was no significant difference between the diabetic groups C, D, and E. (p <0.05)

The amount of drinking water was increased about 5 times in diabetic rats compared to normal rats. There was no significant difference between diabetic group C, D, and E group. There was also no significant increase or decrease from week to week in each group (see Figure 2). (P <0.05)

Feed intake increased 1.5 times in diabetic rats compared to normal rats. There was no significant difference between the diabetic groups C, D, and E (see Fig. 3). (P <0.05)

<3-3> Changes in Blood Glucose Following Administration of Mixed Extracts

Blood sampling to measure blood glucose and insulin levels was performed after 14-15 hours of fasting at 0, 3, and 6 weeks, respectively. At 0 and 3 weeks, the patient was anesthetized with diethyl ether and the blood was collected from the orbital vein. The collected blood was centrifuged at 4 ° C and 3000rpm for 15 minutes to separate the serum, and then stored at -20 ° C until analysis.

As a result, the blood glucose level of all diabetic groups increased significantly at week 0 compared to the normal group (P <0.0001). There was no significant difference in blood glucose levels between group A and group B at weeks 0, 3, and 6. (P0.05) In diabetic group C, D, and E, there was no significant difference in weeks 0 and 3, but there was a significant difference in 6 weeks (P <0.05). Decreased by 4% and decreased by 28%. By week, the group A's blood sugar was 95.9% and the group B's blood sugar was 88.2% and the group A's blood sugar decreased by 65.3% and 49.7% compared to the C group. In particular, at week 6, E group showed blood glucose levels of around 200 mg / dl slightly above normal blood glucose levels in 3 of 6 mice (see FIGS. 4 and 5). As a result, it was confirmed that the mixed extract of the present invention has an excellent effect on reducing blood sugar.

<Example 4>

Measurement of Histopathological Changes in Pancreas Following Administration of Bio-Deer Fruit, Prickly Pear, Prickly Pear Bark, Ginseng, Alder Bark and Oak Bark in Type 1 Diabetic Animal Model

<4-1> Pancreatic Tissue Extraction

 Rats divided into five groups as in Example <3-1> were supplied with a large amount of feed and extract feed for 6 weeks, followed by necropsy. After fasting for 14-15 hours, anesthesia was performed with diethyl ether, and then opened, blood was collected from the abdominal vein, and the pancreas was extracted and fixed in 10% buffered formaline within 48 hours.

<4-2> biopsy

Pancreatic tissue embedded in paraffin was made into 3 micrometer sections and subjected to H & E staining to observe the pancreatic islets of Langerhans.

H & E staining showed that the pancreatic tissues were atrophy in the endocrine glands of the endocrine glands, unlike the exocrine glands, which showed no significant difference from the normal groups in all groups that induced diabetes with STZ. In the diabetic group, the atrophy of Langerhans island was decreased in the group fed the feed containing 0.01% and 0.1% of the extract compared to the diabetic control group, and the number of nuclei was also large (see FIG. 6). From the above results, it was confirmed that the mixed extract of the present invention has an antidiabetic effect due to the anti-atrophy effect of Langerhans island.

<4-3> Immunohistochemical Test

Pancreatic tissue was fixed in 4% paraformaldehyde for 24 to 48 hours to be examined for insulin secretion in the beta cells of pancreatic Langerhans island, washed with running water for 1 hour, and then processed into paraffin-embedded tissue. Section was sliced to 3㎛ thickness and attached to the ProbeOn TM Plus slide for immunohistochemical staining and dried sufficiently at room temperature. After removing paraffin with Xylene and passing through the hydrolysis process, 0.3% It was treated with methanol containing hydrogen peroxide for 20 minutes. After washing with distilled water and PBS, 0.01M Sodium citrate buffer (pH 6.0) was preheated in a pressure cooker in order to restore antigenicity, and then heated at a boiling temperature for 8 minutes. The ice water was cooled to room temperature in a bath, treated with 0.05% TPBS, washed with PBS four times for 10 minutes, and then reacted at room temperature for 1 hour by applying a normal blocking serum containing Avidin D. After washing with PBS, PCNA antibody (SantaCruz, mouse polyclonal IgG, 1: 100), which is a biotin-containing primary antibody, was applied and reacted overnight at 4 ° C. After washing with PBS, reacted with Ni-DAB for 2 ~ 3 minutes to develop color. Wash with PBS, apply secondary antibody attached with Biotin, react for 40 minutes at room temperature, wash with PBS, apply ABC reagent, and 30 The reaction was carried out for a minute. After washing with PBS, apply the normal blocking serum containing Avidin D again and react for 1 hour at room temperature. After washing with PBS, apply the Insulin antibody (SantaCruz, rabbit polyclonal IgG, 1: 200) which is the primary antibody containing Biotin. After reacting overnight at 4 ° C. After washing with PBS, a biotin-attached secondary antibody was applied and reacted at room temperature for 40 minutes, washed with PBS, and then coated with ABC reagent and reacted for 30 minutes. After applying for 40 seconds to 1 minute with DAB solution, the color was developed and then examined by mounting.

As a result, immunostaining using insulin antibody by ABC method showed that Langerhans is generally positive in normal group A and B, while partially stained in diabetic group C, D and E. Compared with C, the diabetic control group, and D and E, there was no significant difference, but the proportion of insulin-positive cells was higher in C group than in C group and D group (see FIG. 7).

As described above, the composition for the prevention or treatment of diabetes mellitus or its complications comprising extracts of the barberry, thorny oak bark, thorny oak bark, ginseng, alder bark and oak bark of the present invention as an active ingredient It is effective in preventing or treating complications. In particular, it has excellent efficacy in preventing or treating type 1 diabetes mellitus, chronic hyperglycemia, atherosclerosis, microangiopathy, kidney disease, heart disease and diabetic retinopathy. In addition, the food composition comprising the extract of the bark fruit, thorn oak bark fruit, thorn oak bark bark, ginseng, alder bark and oak bark of the present invention has an excellent effect on the prevention and alleviation of type 1 diabetes or its complications It can be used to manufacture health foods for patients, which is highly industrially applicable.

Figure 1 shows the results of weight change test of rats with mixed extract in STZ-induced type 1 diabetic animal model. (A to E refer to the group of Table 1).

Figure 2 shows the results of a negative change test of rats by mixed extract in STZ-induced type 1 diabetic animal model. (A to E refer to the group of Table 1).

Figure 3 is a test result of the change in feed intake of rats by the mixed extract in the type 1 diabetes animal model induced by STZ. (A to E refer to the group of Table 1).

Figure 4 shows the results of blood glucose changes in rats by mixed extracts in the STZ-induced type 1 diabetic animal model. (A to E refer to the group of Table 1).

Figure 5 shows the results of the blood glucose change test of rats by the mixed extract in STZ-induced type 1 diabetic animal model. (A to E refer to the group of Table 1).

Figure 6 is a micrograph of the observation results of changes in the Langerhans islet cells of rats by the mixed extract in STZ induced type 1 diabetic animal model. (A to E refer to the group of Table 1).

Figure 7 is an immunohistochemistry test to determine the changes in insulin secretion-positive cells in rats by the mixed extract in STZ-induced type 1 diabetic animal model. (A to E refer to the group of Table 1).

Claims (3)

The ratio of 1 to 10: 1 to 5: 1 to 5: 1 to 5: 1 to 5: 1 to 10: 1, 10 to 1, 10: 1 to 10, on the basis of the dry weight of the wild bark fruit, prickly pear fruit, prickly pear bark, ginseng, alder bark and oak bark A pharmaceutical composition for preventing or treating type 1 diabetes mellitus or complications thereof by using the extract extracted by mixing as an active ingredient. The pharmaceutical composition of claim 1, wherein the complication is selected from the group consisting of chronic hyperglycemia, atherosclerosis, microangiopathy, kidney disease, heart disease and diabetic retinopathy. The ratio of 1 to 10: 1 to 5: 1 to 5: 1 to 5: 1 to 5: 1 to 10: 1, 10 to 1, 10: 1 to 10, on the basis of the dry weight of the wild bark fruit, prickly pear fruit, prickly pear bark, ginseng, alder bark and oak bark Food composition for the prevention or alleviation of type 1 diabetes mellitus or complications caused by the extract extracted by mixing as an active ingredient.

Images