KR20100108031A - A composition having an effect of curing and preventing diabetes mellitus - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing mixture extract of medicinal herb for preventing and treating diabetes is provided to prevent and treat renal failure, edema, foot gangrene, cataract, glaucoma, stroke, and diabetic neuropathy. CONSTITUTION: A pharmaceutical composition for preventing and treating diabetes contains 6-10 weight% of Codonopsis pilosulae radix, 6-10 weigh part% of Salviae miltiorrhizae radix, 4-8 weight% of Pinelliae rhizome, 3-5 weight% of Coptis rhizome, 3-5 weight% of Evodiae fructus, 8-12 weight% of Epimedii herba, 4-8 weight% of Rhei radix et rhizome, 4-8 weight% of Perillae folium, 2-4 weight% of Glycyrrhizae radix, 12-20 weight% of Artemisiae capillaris herba, 10-14 weight% of Alismatis rhizome, 4-6 weight% of Poria, 4-6 weight% of Atractylodis macrocephalae rhizome, 4-6 weight% of Polyporus and 2-3 weight% of Cinnamomi ramulus. The medicinal herbs are obtained by freeze-drying the hot water extract of the medicinal herbs.

Description

Pharmaceutical composition for preventing and treating diabetes {A composition having an effect of curing and preventing diabetes mellitus}

The present invention relates to a pharmaceutical composition for preventing and treating diabetes, and more specifically, made up of ginseng, red ginseng, halved, yellow lotus, sesame oil, yin and yang, rhubarb, lobule, licorice, jinjin, taeksa, baekbokyeong, baekchul, seolyeong and gyeji It relates to a pharmaceutical composition for preventing and treating diabetes, including a mixed extract of the herbal medicine.

Diabetes is a disease in which glucose in the blood is excreted through urine. It is one of the most common non-communicable diseases in modern people and a chronic degenerative disease that does not heal fundamentally. It is caused by insulin action and insulin secretion. Metabolic syndrome, characterized by hyperglycemia, is caused by a variety of causes, ranging from absolute deficiency of insulin to resistance to insulin as pancreatic beta cells are destroyed by autoimmune mechanisms.

Diabetes is a serious disease that causes hyperglycemia to be chronic and causes lipid or protein metabolic disorders as well as glucose metabolism, leading to complications such as retina, kidney, nerve, and cardiovascular system. It can be shortened and is the fourth cause of death in Korea.

Recently, the prevalence of diabetes increased year by year due to the rapid economic development, which is about 5 ~ 10% in Korea. In 2004, 8.3 out of 200 people were diabetic. While the number of diabetics is increasing rapidly, it is difficult to continue and proper treatment is emphasized the importance of the development of diabetes treatment.

Current drug treatments for diabetes include oral hypoglycemic agents such as sulfonyluresa, biguanides, a-glucosidase inhibitors, and thiazolidinediones, insulin therapy, Pancreas transplantation is used, but if all are used continuously, side effects such as hypoglycemia and hepatotoxicity may occur, and it is not a fundamental treatment method.

Therefore, the present inventors have made a thorough research to develop a preventive and therapeutic agent for diabetes using natural ingredients with no side effects, ginseng, sweet ginseng, halved, yellow lotus, sesame oil, yin and yang, rhubarb, lobule, licorice, injin, taeksa, baekbokyeong Confirmed that the mixed extract of the herbal medicine consisting of, baekchul, herb and gyeji has an excellent effect on the prevention and treatment of diabetes mellitus, to complete the present invention.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for preventing and treating diabetes, including a mixed extract extracted from natural herbal medicine having no side effects.

According to an aspect of the present invention, the present invention is 6-10% by weight, 6-10% by weight, Salvia 4-8% by weight, 3-5% by weight of yellow lotus, 3-5% by weight of sesame oil, 8-12 Wt%, rhubarb 4 ~ 8%, leaflet 4 ~ 8%, licorice 2 ~ 4%, phosphorus 12 ~ 20%, talc 10 ~ 14%, Baekbokyeong 4 ~ 6%, Baekchul 4 ~ 6% It provides a pharmaceutical composition for the prevention and treatment of diabetes, comprising a mixed extract of the herbal medicine consisting of%, age 4-6% by weight and gage 2-3% by weight.

In the pharmaceutical composition for preventing and treating diabetes of the present invention, the mixed extract is ginseng 150g, salvia 150g, halved 100g, rhubarb 60g, sewage oil 60g, Yin Yang 160g, rhubarb 100g, lobule 100g, licorice 50g, 300 g of jinjin, 200g, It is preferable that it is a mixed extract of the herbal medicine consisting of 80 g of Baekbokryeong, 80 g of Baekchul, 80 g of Giryeong and 40 g of gyeji.

In the pharmaceutical composition for preventing and treating diabetes of the present invention, the extract is preferably lyophilized hot water extract of the herbal medicine.

Hereinafter, the method of the present invention will be described in more detail step by step.

At this time, if there is no other definition in the technical and scientific terms used, it has a meaning commonly understood by those of ordinary skill in the art.

In addition, repeated description of the same technical configuration and operation as in the prior art will be omitted.

The present invention not only direct symptoms of diabetes, but also ginseng, sweet ginseng, halved, yellow lotus, sesame milk, yin and yang, rhubarb, lobule, licorice, injin, taeksa, baekbokyeong, baekryeok, which can prevent and treat complications arising from the onset of diabetes. It is to develop a pharmaceutical composition comprising a mixed extract consisting of 15 kinds of herbal medicines of age and geji.

In particular, the pharmaceutical composition of the present invention can be prevented and treated diabetes by the effect of controlling the phenomenon of increase in blood sugar and decrease in insulin caused by diabetes, even if it is extracted from natural products for a long time taking, The effect of preventing tissue damage of the pancreas and kidneys can prevent or treat complications due to long-standing diabetes.

At this time, the water used to extract the mixed herbal medicine in the present invention refers to water suitable for drinking standards based on the Ministry of Health, generally water or tap water, such as purified water, sterile water, anion water filtered through a filter or water purifier, etc. , Bottled water and the like can be used. However, the solvent used to extract the active ingredient of the mixed herbal medicine in the present invention is not limited to water, and if using a device that removes to the extent that it is not harmful to the human body, organic solvents such as methanol, ethanol, acetone, diethyl ether, etc. Can be used.

And, in the present invention, 6 to 10% by weight, 6 to 10% by weight, red ginseng 6 to 10% by weight, half to 4 to 8% by weight, yellow lotus 3 to 5% by weight, sewage oil 3 to 5% by weight, 8 to 12% by weight, rhubarb 4 ~ 8% by weight, 4-8% by weight of leaflets, 2-4% by weight of licorice, 15-20% by weight of phosphorus, 10-14% by weight of taxi, 4-6% by weight of Baekbokyeong, 4-6% by weight of Baekbyeong, age 4-6 Weighing by weight and Cultivation 2-3% by weight, which is used in excess of the above-mentioned maximum amount of each Chinese herbal medicine does not use enough other necessary ingredients, if less than the minimum amount of the effective effect of each herbal medicine is reduced This is because the above-mentioned range of use of each herbal medicine is preferable for showing the maximum effect on diabetes. At this time, more specifically, 150 g of ginseng, 150 g of red ginseng, 100 g of halves, 100 g of yellow lotus, 60 g of sesame oil, 60 g of Yin Yang Guk, rhubarb 100 g, lobule 100 g, licorice 50 g, 300 g of phosphorus, 200 g of Taekjang, 80 g of Baeknyeong, 80 g of Baeknyeong, and 80 g and The mixed herbal medicine consisting of 40 g of gage and the hot water extracted by adding water are used.

However, in the present invention, in order to prevent or treat diabetes in addition to water, while fully extracting the active ingredient of the herbal medicine if harmless to the human body using an organic solvent and then may be subjected to a process of removing the organic solvent.

As described above, the extract of the mixed herbal medicine according to the present invention is able to provide a synergistic effect of prevention or treatment while minimizing side effects for the overall symptoms related to diabetes due to the appropriate balance between mutual efficacy as described above.

The present invention provides a pharmaceutical composition for preventing and treating diabetes by applying the mixed herbal medicine extract. At this time, the pharmaceutical composition according to the present invention can be formulated based on the formulation criteria of the Food and Drug Administration (KFDA) into a conventional pharmaceutical formulation.

The mixed herbal extracts may be used by themselves or on acceptable formulations as pharmaceutical formulations, for example microcrystalline cellulose, colloidal silicon dioxide, corn starch and disintegrants, for example corn starch, crospovidone And pregelatinized starch and glidants such as glyceryl behenate and epidermal agents such as Opadry and the like. In addition, the herbal extracts and formulated preparations of the present invention are conventionally mixed, diluted with a pharmaceutically acceptable carrier, or encapsulated in a container-type carrier according to the administration method, dosage form, and therapeutic purpose. .

When the carrier is used as a diluent, powder, granule, injection solution, syrup, solution, tablet for oral administration and parenteral administration using at least one carrier selected from the group consisting of saline, buffer, water, Ringer's solution and ethanol. To formulations such as; However, the carrier of the present invention is not limited to the above carrier. In this case, parenteral administration refers to the administration of the active ingredient via rectal, intravenous, peritoneal, muscle, arterial, transdermal, nasal, inhalation and the like in addition to oral.

The formulation may further include fillers, anti-coagulants, lubricants, wetting agents, flavors, emulsifiers, preservatives, and the like, and may be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. And the dosage of the present invention can be adjusted according to the condition of the patient, the route of administration and the dosage form is not limited and those skilled in the art according to the symptoms can be obviously used within various ranges, In general, in the present invention, it is determined that 100 mg to 200 mg per kg of body weight can be continuously or intermittently administered per day in an experimental effective amount.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Example 1. Preparation of Herbal Mixture Extract of the Present Invention

TABLE 1

Drug name Scientific name and part Working weight (g) Ginseng radices of Codonopsis pilosula (FR.) NANNF
; Codonopsis pilosulae radix 150 Salvia radices of Salvia miltiorrhiza BGE; Salviae miltiorrhizae radix 150 Half rhizome of Pinellia ternata (THUNB.) BREIT. ; Pinelliae rhizoma 100 goldthread rhizome of Coptis chinensis FRANCH. ; Coptis rhizoma 60 Osuyu fructus of Evodia rutaecarpa (Juss.) Benth. ; Evodiae fructus 60 Yin Yang leaves of Epimedium koreanum NAKAI; Epimedii herba 160 rhubarb rhizome of Rheum palmatum L.; Rhei radix et rhizoma 100 Leaflets leaves of Perilla frutescens (L.) BRITT. ; Perillae folium 100 licorice radices of Glycyrrhiza uralensis FISCH. ; Glycyrrhizae radix 50 Injin leaves of Artemisia capillaris THUNB. ; Artemisiae capillaris herba 300 Taxi rhizome of Alisma plantago-aquatica var. orientale SAMUELS.
; Alismatis rhizoma 200 Baikyeongryeong whole fungi of Poria cocos (SCHW.); Poria 80 Whiteness rhizome of Atractylodes macrocephala KOIDZ.
; Atractylodis macrocephalae rhizoma 80 Bad whole fungi of Polyporus umbellatus (PERS.) FRIES. ; Polyporus 80 Cock branches of Cinnamomum cassia PRESL. ; Cinnamomi ramulus 40

To 1,710 g of the mixed herbal medicines prepared according to the weight of the natural herbal medicines prepared in Table 1, the water volume of about 2 times the volume of the herbal medicines was added, heated for 5 hours to extract hot water, and the extract was filtered with gauze to freeze the extract. By drying, the herbal mixture extract of the present invention was prepared. At this time, the yield was 23.7%.

The extract was named WHW extract, and was used as a medicament for effect verification while storing it at 4 ° C.

Experimental Example 1. Anti-diabetic effect of WHW extract through animal experiment

1) Production of diabetes-induced animal model

end. Preparation of Laboratory Animals

Experimental animals were used as an average of 200g, 8-week-old Sprague-Dawley male from Samtaco (Ilsan, Korea). General solid feed (Samyang blended feed animal, Samyang oil and fat feed, Seoul, Korea) and water were supplied during the entire experiment period. The room temperature was 22 ± 2 ℃, the humidity was 55 ± 5%, and the contrast was 12 hours (Day light 06: 00 ~ 18: 00) was maintained at a constant breeding condition until the end of the experiment.

I. Diabetes induction in experimental animals

Streptozotocin (STZ) was immediately dissolved in 0.01 M citrate buffer (pH 4.5), and a single dose of 0.1 ml per 100 g body weight of the experimental animals was administered at a dose of 60 mg STZ / kg (body weight). In order to confirm that diabetes was induced, fasting 12 hours after 72 hours of STZ administration, blood was collected from the tail, and only rats having a blood glucose concentration of 300 mg / dL or more were used for the experiment.

2) Selection of experimental group and administration of WHW extract

The experimental group was adjusted to the average value of blood sugar levels between the groups, saline-control (Saline-Control; Saline-C), Streptozotocin control (Streptozotocin Control, STZ-C) and the WHW-administered group (W100; WHW 100 mg / kg) of the present invention. Divided by. The administration period was 4 weeks, and the Saline-C and STZ-C groups received 0.1 ml of saline per 100 g body weight orally once a day, and the W100 group received oral WHW extract once daily at a concentration of 100 mg / kg. Administered.

3) Weight measurement of experimental animals

Weight change was measured once a week for the duration of the experiment, with body weight at 0 days of administration as the initial body weight and body weight at 28 days as the final body weight.

As a result, as shown in Figure 1, the weight of the week 0 was 276.31 ± 12.56g in the Saline-C group, 273.33 ± 14.53g in the STZ-C group, 283.33 ± 16.67g in the W100 group was similar to all groups appear. On the other hand, at 2 weeks, body weight gained to 310.01 ± 2.66g in the Saline-C group alone, 275.05 ± 1.45g in the STZ-C group and 276.67 ± 23.33g in the W100 group. In addition, the weight of the Saline-C group was increased to 330.0 ± 14.63g at 4 weeks, but the STZ-C group was reduced to 238.0 ± 17.89g and the W100 group was 274.3 ± 31.02g. However, in the last 4 weeks, the W100 group showed a significant (p <0.05) increase in body weight compared to the STZ-C group.

In the STZ-treated group (STZ-C group, W100 group), the initial weight was not increased compared to the normal saline control group because the normal glucose metabolism could not be due to the onset of diabetes due to the damage of the pancreas. In the case of (W100 group), glucose metabolism due to insulin secretion was normally restored due to the antidiabetic effect of WHW extract, and it was confirmed that the body weight was increased compared to the STZ-C group at 4 weeks. Therefore, the anti-diabetic effect on the WHW extract of the present invention was confirmed.

4) Blood collection of experimental animals

Blood was collected from the tail vein after 12 hours of fasting at 0, 7, 14, 21, and 27 days of administration, and serum was separated by centrifugation at 6,000 rpm for 10 minutes. The separated serum was stored at -20 ° C and used as a sample for serum analysis.

In addition, all experimental animals were sedated by injecting 0.2 mg / kg of Rompun (xylazine hydrochlroride, Yuhan) into the abdominal cavity on the last autopsy day, and anesthesia after injecting 1 mg / kg of Ketalar (ketamine hydrochlroride, Yuhan) into the abdominal cavity. 5 ml of blood was collected from the abdominal vein. Serum was separated from the collected blood by centrifugation at 6,000 rpm for 5 minutes within 1 hour.

5) Analysis of ingredients in serum

In order to confirm the antidiabetic effect of WHW extract, the serum of experimental animals collected from each control group was analyzed.

end. Serum Glucose Concentration Analysis

To determine the effect of the WHW extract of the present invention on the symptoms of hyperglycemia that is characteristic of diabetes, glucose concentration in serum was analyzed.

Serum glucose concentration was measured by absorbance at 500 nm using an enzyme reagent (Asan Pharmaceutical, Korea) using a glucose oxidase reaction.

As a result, the serum glucose concentration of the Saline-C group was measured as 116.8 ± 7.85 mg / dl, 443.4 ± 23.76 mg / dl in the STZ-C group, 344.01 ± 33.96 mg / dl in the W100 group, as shown in FIG. It was confirmed that the serum glucose concentration was significantly (p <0.01) decreased by the WHW extract of the present invention compared to the STZ-C group. This means that the WHW extract of the present invention can effectively treat the hyperglycemic symptoms caused by diabetes.

I. Serum insulin concentration analysis

Insulin concentrations in serum were analyzed to determine the effect of the WHW extract of the present invention on the reduction of insulin secretion from the pancreas, a symptom of diabetes.

Serum insulin concentration was measured by absorbance at 450 nm using a rat insulin ELISA kit (Central Laboratory Animal, Japan).

As a result, as shown in FIG. 3, the serum insulin concentration of the Saline-C group was 0.44 ± 0.02ng / ml, the STZ-C group was measured at 0.209 ± 0.007ng / ml, and the W100 group was 0.286 ± 0.021ng / ml. It was confirmed that the serum insulin concentration was significantly (p <0.01) increased by the WHW extract of the present invention compared to the STZ-C group. This means that by using the WHW extract of the present invention can effectively restore insulin secretion from the pancreas, which is reduced due to diabetes, diabetes can be treated.

All. Serum tryglyceride (TG) concentration analysis

The concentration of triglyceride in serum was analyzed to determine the effect of the present invention WHW extract on the increase of triglyceride, a symptom of diabetes.

The concentration of TG was measured by absorbance at 450 nm using TG diagnostic reagent (Asan Pharmaceutical, Seoul) by Kinetic alkaline picrate method.

As a result, as shown in FIG. 4, the TG concentration of the Saline-C group was measured to be 131.3 ± 35.56 mg / dl, the STZ-C group was 226.8 ± 18.28 mg / dl, and the W100 group was 179.2 ± 17.51 mg / dl. It was confirmed that the WHW extract of TG concentration significantly (p <0.01) compared to the STZ-C group. This means that by using the WHW extract of the present invention, diabetes can be cured by reducing the action of insulin in the target organ by reducing triglycerides.

la. Serum Blood Urea Nitrogen (BUN) Concentration Analysis

Continued progression of diabetes is accompanied by damage to the kidneys, a complication of diabetes, which results in increased BUN levels in the blood due to decreased kidney function. Serum BUN concentration was analyzed to confirm the effect of the present invention WHW extract on these symptoms.

Serum BUN concentration was measured by absorbance at 520 nm using Quanti Chrom ^™ Urea Assay Kit (Bioassay systems).

As a result, as shown in FIG. 5, the BUN concentration of the Saline-C group was 29.49 ± 3.85 mg / dl, the BUN concentration of the STZ-C group was 107.9 ± 6.008 mg / dl, and the BUN concentration of the W100 group was 79.68 ± 8.279 mg / dl. As measured by, the concentration of serum BUN by the WHW extract of the present invention was confirmed that significantly (p <0.001) compared to the STZ-C group. This means that the WHW extract of the present invention reduced the symptoms of diabetes due to the diabetic treatment effect, thereby restoring the function of the kidney, thereby reducing the BUN concentration in the blood.

hemp. Serum creatine concentration analysis

If the kidney is damaged due to the continued progression of diabetes, the creatinine concentration in the blood increases due to the decrease in renal function. The serum creatinine concentration was analyzed to confirm the effect of the present invention WHW extract on these symptoms.

Serum Creatinine concentration was measured by absorbance at 510 nm using QuantiChrom ^™ Creatinine Assay Kit (Bioassay systems).

As a result, the serum creatinine concentration of the Saline-C group was measured as 0.55 ± 0.11 mg / dl, 3.34 ± 0.190 mg / dl in the STZ-C group, 1.133 ± 0.347 mg / dl as in Figure 6, The serum creatinine concentration was significantly (p <0.01) decreased by the WHW extract of the present invention compared to the STZ-C group. This means that due to the therapeutic effect of diabetes mellitus of the WHW extract of the present invention, the symptoms of diabetes are alleviated and the function of the kidney is restored, thereby reducing the concentration of creatinine in the blood.

6) Oral glucose tolerance test (OGTT)

When the WHW extract was administered, an OGTT experiment was performed to confirm the change in the absorption efficiency of oral glucose in the body.

After 4 weeks of physiological saline or WHW extract (100mg / ㎏) administered to the animals and fasted for at least 12 hours, distilled water or WHW (W100mg / ㎏) was administered by 1ml each. After 10 minutes, glucose (1 g / kg / mL) was orally administered, and blood was collected from the tail vein at 30, 60, 90, and 120 minutes to separate serum, and the same method as the measurement of glucose concentration in serum. OGTT was performed in the saline control group (Saline-C), STZ control group (STZ-C) and WHW treatment group (W100), respectively.

As a result, in the Saline-C group as shown in Figure 7, glucose concentrations of 106 ± 5.2 mg / dl, 156 ± 11.8 mg / dl, 148 ± 18.4 mg / at 0 minutes, 30 minutes, 60 minutes, 90 minutes, 120 minutes dl, 137 ± 18.2 mg / dl, 118 ± 20.1 mg / dl, and STZ-C group had glucose concentration of 400 ± 24.7 mg / dl, 480 at 0, 30, 60, 90 and 120 minutes. ± 27.9 mg / dl, 441 ± 26.2 mg / dl, 416 ± 28.9 mg / dl, and 421 ± 24.4 mg / dl. The W100 group had glucose concentrations of 407 ± 11.7 mg / dl, 455 ± 17.9 mg / dl, 362 ± 13.6 mg / dl, 353 ± 15.1 mg / dl at 0, 30, 60, 90 and 120 minutes. Measured at 344 ± 19.4 mg / dl it was confirmed that the concentration of glucose significantly (p <0.05) at 60 and 90 minutes compared to the STZ-C group.

7) Hematoxylin & Eosin Tissue Dyeing

When the WHW extract was administered, H & E tissue staining was performed to identify changes in the pancreatic and renal tissues damaged by diabetes.

For 4 weeks, 100 mg / kg of saline or WHW was administered to each experimental animal group, followed by sedation by injection of 0.2 mg / kg of Rompun into the abdominal cavity, anesthesia by injection of 1 mg / kg of Ketalar into the abdominal cavity, followed by an open physiological saline solution. Was perfused to the heart to remove blood, and the pancreas and kidney were excised and fixed in 4% paraformaldehyde solution for one day. The immobilized tissue was allowed to settle on 10-30% sucrose solution and completely embedded on dry ice using OCT compound. The embedded tissue was freeze-dried at -70 ° C, and a continuous section having a thickness of 30-40 µm was prepared using a freezing section. Thereafter, the cells were dyed in Harris hematoxylin solution for 3 minutes, followed by 1% HCl alcohol and Ammonia followed by differential dyeing with eosin for 3 minutes. After dehydration in the order of alcohol concentration (70-100%) again, the stained tissue specimens were mounted using a mounting medium and examined for staining results.

end. Morphological changes in pancreatic tissue

As a result of observing the shape of the pancreatic tissue, in the normal group (Normal-C) as shown in Figure 8, the beta cells secreting islets of Langerhans (inslets of Langerhans) and insulin, which is a normal endocrine organ around red blood vessels, were observed. In the STZ control group (STZ-C), the pancreas was severely destroyed by STZ-induced diabetes, resulting in structural changes, and almost no normal form of Langerhans island was seen. In the WHW-administered group (W100) of the present invention, the histological degeneration was slightly compared to that of the normal group, but the morphology of the pancreas was restored compared to the STZ control group.

I. Morphological changes in kidney tissue

As a result of morphology of the kidney tissue, as shown in FIG. 9, in the case of the Normal-C group, a typical glomerular, distal tuble, and proximal tubule in the cortex part of the cortex was typical. Nephron structures were recognized, and medullary ray consisting of aggregated tubules extending into the cortex was normally observed. However, the STZ-C group showed typical STZ-diabetic nephropathy, including proliferation and necrosis of tubular epithelial cells and fibrosis of tubules, and marked vasodilated atrophy of glomeruli. On the other hand, in W100 group, histological changes of tubules and glomeruli due to STZ were reduced, and morphological characteristics similar to those of normal group were observed.

8) Immunohistochemical Staining for Insulin

When the WHW extract was administered, immunohistochemical staining was performed to confirm the functional change of insulin secretion of beta cells in pancreatic tissue damaged by diabetes.

First, the pancreatic tissue sections were washed three times with PBS and then blocked with PBS containing 5% normal goat serum and 0.3% Triton X-100 for 1 hour at room temperature. The anti-insulin monoclonal antibody diluted 1,000-fold in PBS containing 2% normal goat serum and 0.15% Triton X-100 was reacted overnight at 4 ° C. and washed three times with PBS. It was reacted with biotinylated secondary antibody (goat anti-rat, 1: 1000) for 1 hour at room temperature and washed three times with PBS for 1 hour at room temperature with avidine-biotin-horseradish peroxidase complex (Vectastain Elite). After completing the reaction, the tissue sections were washed three times with 0.1 M Tris buffer (TB, pH 7.4), and then reacted for 2-5 minutes in TB containing 0.004% H ₂ O ₂ and 0.05% DAB (Sigma). After dipping into 100% alcohol and xylene, mounting was carried out using a mounting medium and the results of immunostaining were examined.

As a result, in the Normal-C group, as shown in Figure 10, beta cells secreting insulin in the normal Langerhans islet stained by insulin antibodies, while STZ-C secreted beta cells by STZ and secreted insulin And not stained with insulin antibodies. The functional changes of Langerhans Island due to STZ did not recover as in the normal group, but increased beta cells stained with insulin antibodies by W100 group, and many normal Langerhans Island were observed. Thus, WHW has been shown to increase insulin secretion by protecting beta cells from STZ-induced beta cell damage.

As described above, according to the present invention by using a mixed extract of Chinese herbal medicine consisting of ginseng, red ginseng, halved, yellow lotus, sesame oil, yin and yang, rhubarb, lobule, licorice, injin, taxa, baekbokyeong, baekrye, nyul and gye, Prevents secondary diabetic complications such as coronary artery disease, stroke, diabetic neuropathy, such as renal failure, edema, foot necrosis, cataracts, glaucoma, and macrovascular complications, as well as primary symptoms of diabetes, such as increased levels and decreased insulin levels And treatment.

1 is a graph showing the body weight of each experimental group including the administration group (W100) of the present invention composition (physiological saline control group: Saline-C and Streptozotocin control group: STZ-C) at two week intervals.

Figure 2 is a graph showing the measurement of the glucose concentration in the serum of each experimental group.

Figure 3 is a graph showing the measurement of the insulin concentration in the serum of each experimental group.

Figure 4 is a graph showing the measurement of serum TG (tryglyceride) concentration of each experimental group.

Figure 5 is a graph showing the measurement of blood serum urea nitrogen (BUN) concentration in each experimental group.

Figure 6 is a graph showing the measurement of the serum creatinine concentration in each experimental group.

Figure 7 is a graph showing the measurement of the glucose concentration in the serum of each experimental group through the OGTT experiment.

Figure 8 is a photograph of the pancreas of each experimental group observed through a microscope after staining Hematoxylin & Eosin tissue.

9 is a photograph of the kidney of each experimental group observed through a microscope after staining Hematoxylin & Eosin tissue.

10 is a photograph taken through a microscope after immunohistochemical staining of the pancreas of each experimental group.

Claims (3)

6-10% by weight of ginseng, 6-10% by weight of red ginseng, 4-8% by weight of hwangja, 3-5% by weight of yellow lotus, 3-5% by weight of sewage oil, 8-12% by weight of yin and yang, rhubarb 4-8% by weight of rhubarb 4 to 8% by weight, licorice 2 to 4% by weight, phosphorus 12 to 20% by weight, taxa 10 to 14% by weight, Baekbokryeong 4 to 6% by weight, Baekchul 4 to 6% by weight, age 4 to 6% by weight, and gage 2 Pharmaceutical composition for the prevention and treatment of diabetes comprising a mixed extract of herbal medicine consisting of ~ 3% by weight. According to claim 1, wherein the mixed extract ginseng 150g, Dansam 150g, 100g hwang 100g, rhubarb 60g, sewage oil 60g, yinyangkok 160g, rhubarb 100g, lobular 100g, licorice 50g, injin 300g, Taek 200g, Baekbokyeong 80g, Baek 80g, low age Pharmaceutical composition for preventing and treating diabetes, characterized in that the mixed extract of the herbal medicine consisting of 80g and gage 40g. According to claim 1 or 2, wherein the extract is a pharmaceutical composition for preventing and treating diabetes, characterized in that the lyophilized hot water extract of the herbal medicine.

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