| Diabetic nephropathy(diabetic nephropathy, DN) is one of diabetic microvascular complications, and is the leading cause of end stage renal disease. Active prevention and treatment of early diabetic nephropathy is of great significance to delay the progression.Yi Qi Yang Yin Huo Xue’s prescription summarized by mentor professor Zhang Gengliang according to many years of clinical experience,can play a role in protection of renal and progression in prevention and treatment of diabetic nephropathy.Objective:In this study streptozotocin was administered intraperitoneally for induction of diabetic rat model.Afterwards,the medical intervention was used on diabetic rats, to observe the effect of Yi Qi Yang Yin Huo Xue’s prescription on the blood glucose levels, lipid profiles, renal function, urinary albumin excretion rate of 24 hours, kidney hypertrophy index, and the expression of matrix metalloproteinase-9 on tenal tissue in diabetic rats,and to investigave the mechanism of Yi Qi Yang Yin Huo Xue’s prescription in protection of renal.Method:Sixty healthy male Wistar rats of clean grade,weiting about 230-250 g, were selected and fed with normal diet to adapt for one week. Twelve rats were randomly selected as normal control group(N), the other forty-eight rats were induced diabete through single intraperitoneal injection of 1%streptozotocin citrate buffer(buffer concentration of 0.1mmol/L,p H 4.5),according to 55mg/kg body weight. The normal control group was given the same volume of 1% citric acid buffer by single intraperitoneal injection. After 72 hours, the tail venous blood of rats was collected to test blood glucose,the blood glucose levels were more than 16.7mmol/L, and the water intake and urine output increased obviously, then diabetic rat of models were induced successfully.Forty-eight diabetic rats were randomly divided into there groups:model group(M),Yi Qi Yang Yin Huo Xue’s group(tradition Chinese medicine group,H),medicine irbesartan group(western medicine group,W), Yi Qi Yang Yin Huo Xue unite irbesartan group(combination group,H+W).There were 12 rats in each group.After one week of induction of diabetic rat model,the medicine was given to the rats according to 10 times dose of adult.The Chinese medicine group were given the party 10ml/kg(equivalent to herbs 32.5/kg) by intragastric administration.The western medicine group were given irbesartan tablets suspension 10ml/kg(equivalent to original drug 2.5mg/ml) by intragastric administration daily.Combined group every morning were given equal dose of troditional Chinese medicine, every afternoon given the same dose of irbesartan by intragastric administration. Normal control group and model group were given an equal volume of saline daily. The intervention of medicine was given by intragastric administration for 8 weeks.At the end of the eighth week, 24-hour urine was collected by using metabolic cages and the volum of urine was noted,to test 24-hour urinary albumin excretion rate.After the final intragastric administration,all rats were fasted for 12 hours, without the limit of drinking.Then the rats were anesthetized by being injected intraperitonealy with 2% pentobarbital sodium, acrodding to 20mg/kg body weight.The blood was taken from femoral artery to test blood glucose, renal function, blood lipids.Renal production of specimens were taken for light microscopy morphological changes of renal pathology and immunohistochemical determination of the expression of matrix metalloproteinase-9.Results:1 General situationsRats of normal control group had good mental state.Their eating, drinking and urine were regular and weight grew significantiy, Their fur was of luster and activities were free. Rats in model group were in poor mental state,at the same time,they had more food, drink and polyuria, The weight loss odviousiy and fur became yellow,while activity decreased and reactions were slow. Some of the rats appeared a series of diabetes complications,for example cataract, infection and limb edema. The general situations of treatment groups were more than that of model group.There was one death in each group of irbesartan group and model group because of biting with each other. There was also one death in each group of Yi Qi Yang Yin Huo Xue’s group and model group because of faild gavage.2 The changes of blood glucoseCompared with the normal group,the blood gloucose levels of model group, Yi Qi Yang Yin Huo Xue’s group, irbesartan group,and combined group were significantly higher, and there were significant differences(P﹤0.05); Yi Qi Yang Yin Huo Xue’s group and combined group had certain hypoglycemic effect, blood glucose decreased obviously compared with model group, there were significant differences(P﹤0.05); irbesartan group had no significant hypoglycemic effect compared with model group, there was no significant difference(P﹥0.05).3 The changes of blood lipidsCompared with normal control group,model group, blood lipids of Yi Qi Yang Yin Huo Xue’s group, irbesartan group and combined group increased significantly; Yi Qi Yang Yin Huo Xue’s group and combined group have lipid-lowering effect, compared with model group, there are significant differences(P﹤0.05); but irbesartan group compared with model group there was no significant difference(P﹥0.05).4 The changes of kidney hypertrophy indexThe kidney hypertrophy indexes of Model group,Yi Qi Yang Yin Huo Xue’s group,irbesartan group and combined group were higher than that of normal group, there are significant differences(P﹤0.05);Compared with model group, the kidney hypertrophy indexes of three treatment groupdecreased, with significant difference(P ﹤ 0.05); Compared with Chinese medicine group and irbesartan group, kidney hypertrophy index in combi-ned group decreased significantly(P﹤0.05).5 The changes of renal function and 24 hours urinary albumin excretion rateCombined with normal group, the urea nitrogen and creatinine of model group, Yi Qi Yang Yin Huo Xue’s group, irbesartan group and combined group were significantly higher,there are significant differences(P ﹤ 0.05);three treatment group compared with model group,urea nitrogen and creatinine are lower, there are significant differences(P ﹤ 0.05);Compared with Chinese medicine group and irbesartan group,urea nitrogen and creatinine of combined group decreased significantly(P﹤0.05).24 hours urinary albumin excretion rate of model group increased significantly compared with normal control group, there are significant differences(P﹤0.05); Three treatment group of 24 hours urinary albumin excretion rate of rats compared with model group, accidentally significant difference(P﹤0.05);Compared with Chinese medicine group and irbesartan group, 24 hours urinary albumin excretion rate of combined group decreased significantly(P﹤0.05).6 The changes of renal pathological morphologyNormal control group glomerular structure are completed without volume increasing.Basement membrane and mesangial matrix of glomerular capillary were normal; the volume and mesangial matrixthe of glomerulus in model group are increased.Three treatment group compared with model group, have different degrees of ease, and combined group is most obvious.7 The influence of renal matrix metalloproteinase-9 expressionMMP-9 chemical staining showed immune histochemical method, in normal group, renal glomerulus and renal tubular epithelial cells have very strong expression. Model group glomerulus and tubular of renal showed weak expression of MMP-9, significantly weaker than the normal group. Three treatment group compared with model group, the expression of MMP-9 increased(P <0.05), especially the combined group decreased more obvious.Conclusion:1 Yi Qi Yang Yin Huo Xue’s prescription can reduce the blood glucose,blood lipids,serum creatinine and blood urea nitrogen of diabetic rats, at the same time reduce urine protein,to protect the renal.2 Yi Qi Yang Yin Huo Xue’s prescription can protect the structure and function of the renal by increasing expression levels of MMP-9 in renal ofdiabetic rats,that provides new way for traditional Chinesemedicine treatment of diabetic nephropathy. |
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