| Objective: TO observe prevention and cure effect of TANGSHEN capsule on the renal damage of diabetic rats systematically and explore the best dosage of Astragalus membranaceus in TANGSHEN capsule by animal experiment. At the same time, to do some further investigation on the mechanism of TANGSHEN capsule protecting the kidney, so as to find an effective way for the prevention and cure of diabetic nephropathy (DN), providing strong theoretical and experimental basis for the clinical spread of TANGSHEN capsuleMethods: 1. TANGSHEN capsule were made and divided into three species: TANGSHEN-1 TANGSHEN-2 TANGSHEN-3 according to the different dosage of Astragalus membranaceus in them. 2. Animal models were made. Male Wistar rats, weight 180-210g, were chosen and fed for 5 days, then fasting for 12 hours. Streptozotocin(STZ) ( bought from American Sigma) were dissolved in newly-made 0.1mmol/L citrate buffer(pH 4. 5) to make into 10mg/ml STZ solution, then injected into rats (60mg/kg bodyweight). The levels of blood glucose were determined 3 days after the injection of STZ, and rats with blood glucose levels ≥13.5mmol/L were used as diabetic rats model. In control group the same volume of citrate buffer with lOmg/ml STZ solution were injected 3. Diabetic modelrats were divided into 4 groups: diabetic rats treated with low dosage (Group LDT) ; diabetic rats treated with middle dosage (Group MDT) ;diabetic rats treated with high dosage(Group HDT); diabetic nephropathy rats (Group DM) .there was a control group (Group NC) still. 4. Drugs injected per kg of rats were converted by 6. 25 times as adult. LDT group: 13 rats were injected with TANGSHEN-1 (with the content of Astragalus membranaceus matching 30g used for 60kg adult) qd, 8 weeks; MDT group:13 rats were injected with TANGSHEN-2(with the content of Astragalus membranaceus matching 60g used for 60kg adult) qd, 8 weeks; HDT group:13 rats were injected with TANGSHEN-3(with the content of Astragalus membranaceus matching 120g used for 60kg adult) qd, 8 weeks; DM group and NC group: 13 rats of each group were injected with the same dosage of distilled water, qd,8 weeks. All rats were allowed free access to food and water with 12 hours illumination. 5. Tested item and method: 8 weeks later, the following studies were performed in each group, such as body weight, kidney weight, the amount of 24 hours urine, fasting blood glucose(FBG), trilycyceride(TG) , cholestrol(CHO) , serum creatinine(Scr), urine creatinine, and urine albumin excretory rate(UAER). The index about kidney hypertrophy (ratio of kidney weight to body weight) and creatinine clearance rate corrected by body weight (Ccr) were calculated. The renal tissue of diabetic rats was observed with light microscope after periodic acid-Schiff (PAS) and HE staining, and the results were analyzed semi- quantitatively with the pathological image analysis system. Immunohistochemical technique was used to estimate the expression of intercellular adhesion molecule-1(ICAM-1) in The renal tissue of diabetic rats. The experimental data was demonstrated in means ± standard deviation. The analysis of variance were used to the significancetest by SPSS 11.5, a statistic software.Result: Results:1.All the improvement of FBG> FIM% mAlb> the morphology of kidney and the interecellular adhesion molecule-1 (ICAM-1) in the group of LDT > MDT ^ HDT were of statistical significance, but still not the normal level. 2. The improvement of mAlb^ the morphology of kidney and the interecellular adhesion molecule-1 (ICAM-1) in HDT group was most significant. Between LDT and MDT group, there wasn' t significant difference.3. Cr^ Ccr in HDT group were of significant difference compared with the DM group, but no difference compared with the NC group. MDT and HDT group were of statistical difference compared with the NC group,but no difference compared with the DM group.Conclusion:1.All the three groups of TANGSHEN capsule have protection effect on the renal damage of diabetic rats, while the HDT group is the best. 2. All the three groups of TANGSHEN capsule can improve the glycometabolism of diabetic rats, lower the blood glucose and serum fructose, but can' t control the blood glucose effectively. 3. All the three groups of TANGSHEN capsule can improve the fat metabolism effectively, while the HDT group is the best. 4. All the three groups of TANGSHEN capsule can improve the mAlb, while the HDT group is the best. 5. The HDT group can improve the creatinine and creatinine clearance rate in diabetic rats evidencely , while the LDT and MDT group can not. 6. ICAM-1 has increased positive rate in the kidney of diabetic rats, may be concerned with the course of renal damage of diabetic rats. 7. All the three groups of TANGSHEN capsule can lower the expression of ICAM-1 in the kidney of diabetic rats, especially the HDT group, while the LDT and MDT group have worse effect.
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