KR20080011921A - Compositions for treating autoimmune disease containing extracts of salviae miltiorrhizae radix - Google Patents

Abstract

An extract of Salviae Miltiorrhizae Radix is provided to decrease the total IgG amount of anti-dsNDA, an important marker of an auto-immune disease, and the amount of IFN-gamma, thereby being very useful for treating the autoimmune diseases. A pharmaceutical composition for treating autoimmune diseases comprises an extract of Salviae Miltiorrhizae Radix, where the autoimmune diseases is rheumatoid arthritis, systemic lupus erythematosus(SLE), polyneuritis, type I diabetes, inflammatory bowel disease or scleroderma.

Description

Compositions for treating autoimmune disease containing extracts of Salviae Miltiorrhizae Radix}

Figure 1 shows a comparison of body weight between the group and the control group in NZB / w F1 mice.

2 shows the comparison of proteinuria and urine protein levels between the group treated with the control group and the control group in NZB / w F1 mice.

Figure 3 shows the comparison of the serum level of anti-dsDNA IgG between the mono-dose group and the control group in NZB / w F1 mice.

Figure 4 shows the comparison of the serum level of anti-dsDNA IgG1 between the mono-administration group and the control group in NZB / w F1 mice.

Figure 5 shows the comparison of the serum level of anti-dsDNA IgG2a between the short-term administration group and the control group in NZB / w F1 mice.

Figure 6 shows the comparison of IFN-γ expression level between the control group and the control group in the spleen lymphocytes of NZB / w F1 mice.

Figure 7 shows the comparison of IL-4 expression level between the control group and the control group in the spleen lymphocytes of NZB / w F1 mice.

Figure 8 shows the histological examination results between the group and the control group in the renal tissue of NZB / w F1 mice. A is the height of 27-week-old NZB / w F1 rats, and B is the short osmotic group.

Autoimmune disease is a disease caused by the body's immune function attacking itself. In order to treat autoimmune disease, the immune function that protects our body must be suppressed. have. In addition, autoimmune diseases are formed over a long period of time, the symptoms are chronic and usually cause permanent damage of the organ is a common example, the reality is that there is little way to cure. While knowledge of autoimmune diseases has evolved over the past 20-30 years, the exact mechanisms of production, the identity of autoantigens and regulatory genes are still unclear. Autoimmune diseases can be broadly divided into organ-specific diseases and systemic diseases.

Organ-specific autoimmune diseases occur as a result of an immune response to organ-specific antigens and can occur in almost every organ in our body. Systemic autoimmune diseases are caused by immune responses to antigens expressed throughout the body rather than by an immune response to any particular cell. These systemic autoimmune diseases can also cause disease selectively in specific organs (see http://irc.ulsan.ac.kr/immunity/index.htm, accessed Apr. 11, 2006, 4:30 pm).

Examples of autoimmune diseases include i) rheumatoid arthritis, in which the immune system attacks tissues of various joints, and ii) autoimmunity of the central nervous system induced by T cells. Multiple sclerosis (MS), which can lead to blindness, paralysis, and premature death, is caused by the destruction of immune cells in the insulin-producing cells of the pancreas, and the MHC gene is important for immune mediating or type 1 diabetes. Mediated or Type 1 Diabetes Mellitus, ⅳ) Inflammatory Bowel Diseases, a disease caused by the immune system attacking the intestine, ⅴ) Scleroderma, which induces thickening of the skin or blood vessels ((). Systemic autoimmunity is accompanied by symptoms such as deep fatigue, rash, joint pain, and, in severe cases, systemic redness, which can damage the kidneys, brain, lungs, etc. And the like sex lupus (Systemic Lupus Erythematosus, SLE) (see http://home.inje.ac.kr/~lecture/immunobiotech/ch6/6autoimmunity.htm, 2006. 04. 11. 16:30 connection).

On the other hand, Salviae Miltiorrhizae Radix (SMR) is a perennial herb belonging to Sun-Hwa family (Lacaceae, Labiatae), which has the tinnitus of red ginseng, wild ginseng, shepherd's milk and horse breeding. This medicine is effective for the treatment of symptomatic such as hemoglobin, hematopoietic pulmonary disease, abundance of rain, infertility, cardiovascular system, blood death, etc. useful. In addition, although the salviae are known to have pharmacological effects such as increased coronary blood flow, lowering blood pressure through peripheral vasodilation, decreased serum lipid, antibacterial action, hypoglycemia, stabilization, analgesic effect, None has been used to treat SLE

Therefore, the inventors of the present invention, after administering the ginseng to lupus-induced NZB / w F1 (New Zealand Black / White F1) mouse model, body weight and proteinuria, kidney and liver function test, blood test, lymphocyte test in spleen, ELISA (ELISA) Cytokine expression, anti-dsDNA antibody expression level, and histological analysis of kidney were used to examine whether the ginseng effectively affects liver and kidney disorders and immune function. It was confirmed that the effect on the treatment of autoimmune diseases and completed the present invention.

An object of the present invention is to provide a use of the extract as a therapeutic agent for autoimmune diseases.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

The first aspect of the present invention relates to a pharmaceutical composition for treating autoimmune disease, comprising an extract of Salvia ginseng. More specifically, the autoimmune disease relates to a pharmaceutical composition, characterized in that the systemic lupus erythematosus, rheumatoid arthritis, multiple neuritis, type 1 diabetes, inflammatory bowel disease and scleroderma.

In the present invention, rheumatoid arthritis refers to an inflammatory disease accompanied by pain, swelling, and joint stiffness due to symmetry of arthritis by the immune system attacking tissues of various joints. Osteoarthritis of rheumatoid arthritis is a degenerative disease and is an inflammatory disease caused by joint decoction. Treatment is an inflammation of the joint.

In the present invention, multiple neuritis (Multiple Sclerosis) is a central nervous system autoimmune induced by T cells, most of the relatively normal life, but can lead to severe blindness, paralysis, premature death Refers to autoimmune diseases.

 In the present invention, type 1 diabetes mellitus (Immune-Mediated or Type 1 Diabetes Mellitus) is an autoimmune disease caused by the destruction of the pancreatic insulin-producing cells by immune cells. Symptoms (fatigue, frequent urination, thirst, acute confusion) are present. Severe diabetes can lead to kidney damage, blindness, limb amputation, and death.

Inflammatory Bowel Diseases in the present invention are diseases caused by the immune system attacking the intestine, and induce symptoms such as diarrhea, vomiting, dizziness, abdominal cramps, and pain.

In the present invention, sclerosis (Scleroderma) is a disease that induces the hardening of the skin or blood vessels (thickening), loss of motor function, shortening of breath, rarely lead to kidney, heart, lung failure.

Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

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

Suitable dosages of the pharmaceutical compositions of the present invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and response to response of the patient, Usually a skilled practitioner can easily determine and prescribe a dosage effective for the desired treatment or prophylaxis.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporating into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or an aqueous medium, or may be in the form of extracts, powders, granules, tablets or capsules, and may further include a dispersant or stabilizer.

The second aspect of the present invention relates to a food composition effective for autoimmune diseases comprising the extract of Salvia mildew. More specifically, the autoimmune diseases are systemic lupus erythematosus, rheumatoid arthritis, multiple neuritis, type 1 diabetes mellitus, inflammatory bowel disease and scleroderma.

Food containing the extract of the present invention may include ingredients that are commonly added at the time of food production. For example, when the beverage is prepared, in addition to the plant extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid and / or juice may be further included.

On the other hand, the functional food composition of the present invention includes components that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when manufactured with a drink, it may further include citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract, etc., in addition to the Dansam extract of the present invention. In view of easy access to food, the food of the present invention is very useful for the treatment or prevention of neurological diseases, for the treatment or prevention of diseases caused by oxidative stress, and for improving cognitive function.

In the present invention, NZB / NZW F1 mice produce a part of autoantibodies and are widely used as a research model for human SLE disease. NZB / NZW mice are born normally clinically but show symptoms of hemolytic anemia within 2-3 months. Anti-erythrocytic antibodies, antinuclear antibodies, lupus cells, and immune complex tests are all positive.

In the present invention, GOT (AST) and GPT (ALT) is an amino acid synthase that exists in the organs, including the liver, which has a certain level in the blood even by normal cell destruction, but when the liver and certain organs are damaged, a large amount of cells are destroyed and eventually These enzymes leak out of the cell, causing the enzyme to rise.

In the present invention, the total protein refers to the sum of various proteins present in the serum, and the nutrition of the body can be known. Total protein is somewhat increased in case of blood concentration due to dehydration, but has little clinical significance and rarely, but increases in pathology in multiple myeloma and decreases in cases of malnutrition, chronic infection, liver cirrhosis, nephrotic syndrome and chronic inflammation. Creatinine is present in most muscles and is excreted through the kidneys. Therefore, in the case of kidney disease such as kidney failure, nephritis, high protein diet, tissue collapse and gastrointestinal bleeding increases in blood, and also increases in muscle diseases. It is somewhat decreased in liver disease such as cirrhosis but has no clinical significance. Blood Urea Nitrogen (BUN) increases with creatinine in kidney disease and decreases in liver disease such as cirrhosis, but has no specific clinical significance. Be careful not to eat high-protein meals, tissue breakdown, gastrointestinal bleeding, dehydration increases because it is necessary to discriminate.

In the present invention, interferon (IFN, Interferones) is divided into three groups of IFN-α, IFN-β, IFN-γ and IFN-γ is mainly produced by activated T cells. The combination of interferon and receptor results in a variety of effects. In particular, IFN-γ has an immunosuppressive effect and increases antigen-providing and activity of macrophages, NK cells, and cytotoxic T lymphocytes. Another important function is that interferon increases the expression of MHC molecules on the cell surface. MHC I molecule expression is increased by all interferons, while MHC II molecules are induced by IFN-γ. IL-4 antagonizes IFN-γ secreted by Th1 cells, which promotes functional cellular immune responses, and acts as an immunotherapeutic agent for autoimmune diseases (type 1 diabetes, rheumatism, multiple sclerosis, etc.) caused by excessive Th1 responses. It has been confirmed that it is possible to inhibit the growth of malignant tumors (plasmacytoma, mammary adenocarcinoma, transformed fibroblast cell lines, a melanoma cell line, sarcoma cell line, etc.) which are refractory diseases. Safety and effectiveness have been confirmed.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Measurements presented in all experiments were expressed as mean ± standard error of mean (SEM), and statistical analysis was performed by Student's t-test. *, P <0.05 (n = 8)

Example 1 Identification of Effect of Salvia Miltiorrhiza on Body Weight of NZB / w F1 Mice

(1) materials and methods

1) Experimental Animal

The experimental animals used in the present invention were 6 weeks old NZB / w F1 (New Zealand Black / White F1) magnetic mice, which were managed under sterile conditions and purchased from Central Experimental Animals (Seoul, Korea). . The feed was purchased from Purina Co., Ltd. (Gyeonggi-do,) and the sterilized distilled water was used. NZB / w F1 mice were kept in general lighting, 22 ± 1 ° C, and 55 ± 5% of humidity while feeding and drinking enough to feed freely for three months and used as experimental animals.

2) Preparation and Administration of Samples

Salviae (Salviae Miltiorrhizae Radix: SMR) used in the present invention was purchased from (Sunten Pharmaceutical Co. Lot No. 102401, Taipei, Taiwan). 0.6 g of salvia and 0.32 g of starch (yield: 66%) were mixed per 1 g. Thus, 66.7 mL of water was added per 1 g of salvia, and extracted at room temperature for 24 hours. The collected extract was centrifuged at 3000 rpm for 10 minutes and the supernatant was filtered using filter paper (whatman, No. 2, Portland, OR, USA) and used for the experiment. In the NZB / w F1 mice, the control group was administered water, and in the group of the group administered with salvia, 1% salvia extract was used orally for 15 weeks.

3) measurement of weight

NZB / w F1 mice orally administered with 1% Salvia Miltiorrhiza for 15 weeks were weighed using an animal laboratory scale (OHAUS corporation, Pine Brook, NJ).

(2) results

As described above, after oral administration of 1% salvia extract to NZB / w F1 mice at 3 months of age for 15 weeks, it was confirmed that there was almost no difference between the control and the body weight ( FIG. 1 ).

<Example 2> Confirmation of the proteinuria reducing effect of salvia

(1) materials and methods

Prepare experimental animals and samples as in Example 1 (1) -1), 2),

After 15 weeks of oral administration of 1% salvia extract to NZB / w F1 mice, urine was collected and proteinuria was measured according to the manufacturer's protocol on URiSCAN test paper (Youngdong Pharmaceutical Co., Ltd., Gyeonggi-do, South Korea). . 10-29 ug / mL is 1, 30-99 ug / mL is 2, 100-299 ug / mL is 3, 300-999 ug / mL is 4, 1000 ug / mL based on the color scheme provided by the manufacturer The results were analyzed by dividing the rating into five.

(2) results

As a result of measuring the proteinuria to confirm whether a dysfunction occurred due to glomerular damage, the group treated with the mononuclear group was significantly reduced compared to the control group ( FIG. 2 ). Most of the patients with SLE develop immunoglobulin deposition in the glomeruli, which is a major cause of lupus nephritis, and half of the patients show proteinuria. Urinalysis is the simplest and most commonly used test to diagnose lupus nephritis. The decrease in proteinuria is thought to be due to an improvement in lupus.

Example 3 Confirmation of Effect of Salvia Miltiorrhiza on Liver Function

(1) materials and methods

  Experimental animals and samples were prepared as in Example 1 (1) -1) and 2), and NZB / w F1 mice 3 months old were orally administered with 1% salvia extract for 15 weeks. Blood was collected. The collected blood was left at room temperature for 1 hour and centrifuged at 3000 rpm for 5 minutes. The separated serum was stored at -20 ℃ was used for the experiment. The separated serum was analyzed using DRI-CHEM 3500s (Fuji Photo Film Co., Ltd, Kamagwa-ken, Japan).

(2) results

 GOT and GPT were examined to confirm hepatocellular damage, bile duct stagnation, or drug-induced enzyme increase. The short osmotic group did not show any significant difference compared with the control group.

Table 1 . Comparison of GOT and GPT Concentrations in NZB / w F1 Mice

Control Salmon Administration GOT (U / l) 65.0 ± 10.63 58.0 ± 18.58 GPT (U / l) 20.33 ± 5.93 17.60 ± 2.14

Example 4 Confirmation of Effect of Salvia Miltiorrhiza on Kidney Function

(1) materials and methods

Experimental animals and samples were prepared as in Example 1 (1) -1) and 2), and 3% old NZB / w F1 mice were orally administered with 1% salvia extract for 15 weeks. As in (1), the blood was collected and the serum was separated and analyzed using DRI-CHEM 3500s (Fuji Photo Film Co., Ltd, Kamagwa-ken, Japan).

(2) results

 BUN, creatinine, and total protein were examined to determine the status of renal impairment and the extent of the disorder. As a result, the test group decreased in all groups compared with the control group, but it was not significant.

Table 2 . Comparison of Total Protein, Creatinine, and BUN Concentrations in NZB / w F1 Mice.

Control Salmon Administration Total protein (g / dl) 4.386 ± 0.16 4.40 ± 0.16 Creatinine (mg / dl) 0.314 ± 0.03 0.280 ± 0.02 BUN (mg / dl) 31.51 ± 6.91 26.52 ± 2.94

Example 5 Effect of Salvia Miltiorrhiza on Hematology

(1) materials and methods

Experimental animals and samples were prepared as in Example 1 (1) -1) and 2), and 3% old NZB / w F1 mice were orally administered with 1% salvia extract for 15 weeks. Hematology analyzer HEMAVET system, Drew Scientific Inc, Dallas, TX , USA).

(2) results

The test results using the hematological analyzer as shown above are shown in Table 3, and the distribution rate of the blood cells was almost insignificant.

Table 3 . Hematological Comparison in NZB / w F1 Mice

Control Salmon Administration White blood cell (K / ul) 1.528 ± 0.24 1.487 ± 0.24 Red Blood Cell (M / ul) 7.959 ± 0.33 7.510 ± 0.33 Lymphocytes (%) 84.80 ± 0.92 86.60 ± 1.60 Neutrophil leukocytes (Neutrophil,%) 8.614 ± 0.93 7.753 ± 1.50 Monocyte (Monocyte,%) 5.374 ± 0.79 6.183 ± 0.87 Eosinophil (%) 0.880 ± 0.24 0.715 ± 0.15 Basophils (%) 0.247 ± 0.07 0.36 ± 0.11

Recently, a method of treating lupus with disease by transplantation using autologous hematopoietic stem cells has been studied. The method is to remove the pathological lymphocytes completely, and then replace them with the newly produced lymphocytes from the transplanted autologous hematopoietic stem cells. Salviae extract according to the present invention increased the amount of lymphocytes, it is not known whether the increase is an increase in pathological lymphocytes, lymphocytes performing a normal immune function. However, the amount of monocytes, neutrophils and eosinophils was reduced. The relationship between lupus disease with increasing and decreasing lymphocytes requires further study.

Example 6 Effect of Salvia Miltiorrhiza on Anti-dsDNA Antibody Expression

(1) materials and methods

1) Prepare experimental animals and samples as in Example 1 (1) -1) and 2), and after oral administration of 1% Salvia extract for 15 weeks to NZB / w F1 mice 3 months old, As in Example 3 (1), serum was isolated from the blood obtained by heart blood collection.

2) Determination of the expression level of anti-dsDNA antibody using ELISA

In order to measure the anti-IgG, IgG1, IgG2a expression levels in isolated herpes, the protocol of mouse IgG, IgG1, IgG2a ELISA Quantitation kit (ELISA Quantitation kit, Bethyl Laboratory, Montgomery, TX, USA) was used for calf thymus. ) Dilute DNA (Sigma, Louis, MO, USA) to 100 ug / mL with coating buffer (0.05M Carbonate-Bicarbonate, pH 9.6), dispense 100 μl into 96-well plates and coat for 1 hour at room temperature. It was. The coated plate was washed three times with washing buffer (50mM Tris, 0.14M Nacl, 0.05% Tween-20), and then 200 µl of blocking solution (50mM Tris, 0.14M Nacl, 1% BSA, pH 8.0) was added. After aliquoting per well, the cells were blacked for 30 minutes at room temperature. Again washed three times with the wash buffer, and 100 μl of standards and samples were dispensed for 1 hour at room temperature. Washed five times with wash buffer and 100 μl of HRP Detection antibody was added to each well. After 1 hour of reaction at room temperature, the solution was washed 5 times with washing buffer, and then 100 μl of substrate solution (Substrate Solution, TMB Substrate Reagent; Pharmingen, BD Bioscience, San Diego, CA, USA) was added to each well. After reacting for 30 minutes in a dark room temperature, 100 μl of 2N H ₂ SO ₄ was added and then read at 450 nm with a microplate reader (Micropalte reader, Molecular Devices, Sunnyvale, CA, USA) within 30 minutes.

(2) results

As measured by the Eliza method as described above, the anti-dsDNA total IgG was significantly decreased and the anti-dsDNA IgG1 was decreased, but not significant. In contrast, anti-dsDNA IgG2a increased significantly ( FIGS. 3, 4, 5 ).

Anti-dsDNA antibodies are antibodies to DNA, an important constituent in our body, and they play a decisive role in lupus diagnosis because they only appear in lupus patients. In addition, anti-dsDNA antibodies are often used as an indicator of lupus treatment response in hospitals because the higher the lupus, the higher the levels, and the lower the lupus, the lower the levels. Therefore, the salviae extract of the present invention is shown to be effective in lupus disease.

Example 7 Effect of Salvia Miltiorrhiza on Cytokine Expression in Spleen Lymphocytes

(1) materials and methods

1) Prepare the experimental animals and samples as in Example 1 (1) -1), 2), and after oral administration of 1% salvia extract for 15 weeks to NZB / w F1 mice 3 months old, spleen Extract the.

2) preparation and culture of spleen lymphocytes

The spleens of the isolated NZB / w F1 mice are crushed with a sterile syringe and then filtered with a cell strainer (BD Bioscience, San Diego, CA, USA). Into homogenized splenocytes, 5 ml of palm rise (PharM Lyse, BD Bioscience, San Diego, CA, USA) was added to remove red blood cells and allowed to react for 5 minutes. 5 ml of medium is added to the tube in which the cells are suspended, centrifuged at 1,000 rpm for 10 minutes, and the supernatant is removed. The remaining cell pellet was suspended in 1 ml of medium and stained with trypan blue to measure the number of cells.

The isolated spleen lymphocytes were seeded into 24-well plates at a concentration of 2 × 10 6 cells / mL in 24-well plates. 2 μg / mL anti-CD3e (clone: 145-2C11, BD Bioscience, San Diego, CA, USA) coated spleen lymphocytes on coated plates and were then stimulated with 2 μg / ml anti-CD28 (clone: 37.51, BD Bioscience, San Diego, Calif., USA). The mixture was incubated for 48 hours at 37 ℃, 5% CO ₂ incubator (Nuaire, Plymouth, MN, USA) and then obtained a supernatant and stored in -20 ℃ was used in the experiment.

3) Measurement of cytokine expression using ELISA

In order to measure the expression level of IFN-γ and IL-4 in supernatant cultured spleen lymphocyte cells of NZB / W F1 mice, OptEIA mouse IFN-γ set and OptEIA mouse IL-4 set (BD Bioscience, San Diego, CA) , USA) protocol, but the capture antibody (anti-mouse IFN-γ or IL-4) is diluted with coating buffer (0.1M Carbonate, pH 9.5) and dispensed 100μL in 96-well plate and then 4 ℃ Coated overnight at. The coated plate was washed three times with washing buffer (PBS / Tween-20 0.05%), followed by dispensing the assay diluent (Assay Diluent, BD Bioscience, San Diego, Calif., USA) at 200 μl / well, then at room temperature. Blocking for time. Again washed three times with the wash buffer and the standard or sample was dispensed each 100μL and reacted for 2 hours at room temperature. After washing 5 times with washing buffer and adding 100 μl of each working detector (Working Detector, detection antibody + avidin-HRP) to each well, the reaction was performed at room temperature for 1 hour and then washed 10 times with washing buffer. 100 μl of each solution (Substrate Solution, TMB Substrate Reagent; Pharmingen, BD Bioscience, San Diego, Calif., USA) was added to each well. After reacting for 30 minutes in a dark place at room temperature, 50 μl of 2N H ₂ SO ₄ was added and within 30 minutes, 450 nm / 570 with a multi-enzyme immunoassay (absorber, Microplate Reader, Molecular Devices, Sunnyvale, CA, USA). Read at nm.

(2) results

The cytokine expression level measured by the above method was significantly decreased in IFN-γ but not significantly increased in IL-4 compared to the control group compared with the control group ( FIGS. 6 and 7 ).

IFN-γ plays a role in inducing remission in rheumatoid arthritis patients, but it is known to exacerbate the disease in SLE patients and SLE mice. The main mechanism is known to accelerate the production of IgG2a and accelerate glomerular disorder. IL-4 is a cytokine induced by Th2 and mainly secretes IgG1 and IgE and is increased in SLE patients. In case of IFN-γ, which is involved in IgG2a conversion, the saliva extract was significantly reduced compared to the control group, indicating that it was effective in lupus disease. On the other hand, the salviae extract according to the present invention, IL-4 involved in IgG1 conversion increased in comparison with the control group in the saliva extract administration group, but was not significant (?).

Example 8 Checking the Effect of Salvia Miltiorrhiza on Kidney

 (1) materials and methods

Experimental animals and samples were prepared as in Example 1 (1) -1) and 2), and NZB / w F1 mice aged 3 months were orally administered with 1% salvia extract for 15 weeks. After sacrifice, kidney tissue was extracted, washed with saline, fixed in 10% neutralized formalin, washed with water, dehydrated, and fixed with paraffin. The fixed tissue was divided into 4 μm thick and PAS stained using a protocol of PAS (PAC, Periodic Acid-Schiff staining system, Sigma, ouis, MO, USA).

(2) results

As a result of the above experiments, it was confirmed that the dan-sam administered group had less glomerular damage and no invasion of macrophages compared to the control group (FIG. 8).

In Figure 8 , A is the kidney of the control group, the short and wide arrow is a hardened glomeruli, the long arrow shows the surface vascular leukocyte infiltration, B is the kidney of the short-term administration group, two short arrows represent the glomeruli.

It was stained with PAS (Periodic acid-Schiff hematoxylin stain). Magnification × 400.

In the present invention, the proteinuria level was measured after oral administration of the salvia extract to NZB / W F1 mice, and significantly decreased proteinuria, increased lymphocytes, and decreased anti-dsDNA total IgG, an important measure of lupus disease, in the administration group. I was.

In addition, as a result of examining the effect of Salvia extract according to the present invention on cytokine expression in spleen lymphocytes of NZB / W F1 mice, the group treated with Salvia extract was significantly decreased compared to the control group in IFN-γ involved in IgG2a conversion. It has been shown to be effective in the disease, and since it shows an effect of inhibiting the damage of glomeruli and invasion of macrophages, it can be usefully used as a treatment for lupus.

 Salviae extract according to the present invention reduced the amount of anti-dsDNA total IgG, an important marker of autoimmune disease, and decreased the amount of IFN-γ. Therefore, the extract is expected to be useful in the treatment of autoimmune diseases such as rheumatoid arthritis, polyneuritis, type 1 diabetes, inflammatory bowel disease, and scleroderma.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (15)

Pharmaceutical composition for treating autoimmune disease comprising extract of Salvia. The pharmaceutical composition for treating systemic lupus erythematosus according to claim 1, wherein the autoimmune disease is systemic lupus erythematosus (SLE). The pharmaceutical composition for treating rheumatoid arthritis according to claim 1, wherein the autoimmune disease is rheumatoid arthritis. The pharmaceutical composition for treating multiple neuritis according to claim 1, wherein the autoimmune disease is multiple neuritis. The pharmaceutical composition for treating type 1 diabetes mellitus according to claim 1, wherein the autoimmune disease is type 1 diabetes mellitus. The pharmaceutical composition for treating inflammatory bowel disease according to claim 1, wherein the autoimmune disease is an inflammatory bowel disease. The pharmaceutical composition for treating scleroderma according to claim 1, wherein the autoimmune disease is scleroderma. An effective food composition for autoimmune diseases comprising the extract of Salvia. The food composition effective for systemic lupus erythematosus according to claim 8, wherein the autoimmune disease is systemic lupus erythematosus. The food composition effective for rheumatoid arthritis according to claim 8, wherein the autoimmune disease is rheumatoid arthritis. 9. The food composition effective for multiple neuritis according to claim 8, wherein the autoimmune disease is multiple neuritis. The food composition effective for type 1 diabetes, characterized in that the autoimmune disease is type 1 diabetes. The food composition of claim 8, wherein the autoimmune disease is an inflammatory bowel disease. [Claim 10] The food composition of claim 8, wherein the autoimmune disease is scleroderma. A food composition effective for systemic lupus erythematosus comprising the therapeutic agent for autoimmune diseases of claim 8.

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