| Objective: In this experiment,We first established diabetes mellitus (DM) animal model, Then development for diabetic nephropathy (DN), and carried on the intervention treatment using the Yiqi Yangying Huoxue compand Chinese medicine to the model.Through observed experimental animal's fasting blood glucose, 24 hour urinary protein quantitative,serum fructosamine, blood lipids, renal function, pathomorphology chang of kidney, renal cortex in advanced glycation end products (AGEs) and its receptor mRNA (RAGE-mRNA) expression.We researched Yiqi Yangyin Huoxue compound Chinese medicine's therapeutic action to DN and the possible mechanism,and provided the experiment foundation and the theory basis for its clinical application.Methods: Clean healthy male SD rats 40 and weighing 200g±20g, a week normal diet feeding adaptation, metabolic cage urine detection check urine sugar and urine protein were negative, 10 were randomly drawn for the normal control group (normal group , Normal), the remaining 30 by 60mg/kg intraperitoneal injection of streptozotocin chain in urine (STZ), 72h after the blood afterpeak measured blood sugar, urine sugar test from urine. Blood glucose≥16.7mmol/L, positive urine sugar, urine protein-positive Normal urine is greater than 150 percent as a group into a mold. Mode will become the rats were randomly divided into: the model group of diabetic nephropathy (DN-Modol group, Model), the treatment group of compound Chinese medicine (Chinese herbal compound group, ZY), benazepril treatment group (Benazepril group, Benazepril), n = 10. Each group did not give any Western medicine hypoglycemic. Treatment of each group into a module in one week after the start by 20 times the adult dose administration, the normal group and model group, the corresponding amount of distilled water given orally (1ml/100mg), in each group were administered for 8 weeks. In the first eight weeks using metabolic cages to collect 24-hour urine, detection of a 24-hour urinary protein (urinary protein, Upro) quantitative. After administration, the last time, eight hours of fasting, abdominal aortic blood, serum, detection of fasting blood glucose (fasting blood glucose, FBG), blood urea nitrogen (blood urea nitrogen, BUN), creatinine (serum creatinine, Cr), blood total cholesterol (total cholesterol, TC), triglyceride (triglyceride, TG), serum fructosamine (FMN). Aseptic check the kidneys, remove the envelope calculation of removal of kidney hypertrophy index weighing, taking a small piece of kidney tissue fixed, formalin-fixed for observation of renal pathological changes and immunohistochemical detection of the expression of AGEs in renal cortex; liquid nitrogen fixed by in the RT-PCR detection ofthe expression of RAGE-mRNA. Blood and urine biochemical parameters are used fully automatic biochemical analyzer determination. (1ml/100mg), in each group were administered for 8 weeks. In the first eight weeks using metabolic cages to collect 24 hour urine, detection of a 24 hour urinary protein (urinary protein, Upro) quantitative.Results:1 The general experimental ratsNormal diet and normal rats, the eyes of God, have reacted swiftly and wear tight hair, muscle fullness, stool into granular wheat, normal urine. The remaining 3 groups after STZ injection, both obvious and more food and more drink polyuria symptoms appear gradually makethemapathetic , unresponsive,little moves hugs the group. In The last week of the experiment, compared with the normal model rats significantly small size, prominent bones, less muscle, the symptoms in order to model the most significant.2 Body weight of rats in each group, right kidney weight, hypertrophy index (kidney weight / body weight) in comparisonModel group, Benazepril group, Chinese group than the normal weight group was significantly lighter side (P <0.01), and model the three body weight between the groups no significant difference (P> 0.05); normal model group and the right kidney weight of rats in each group no significant difference (P> 0.05); the model group, Benazepril group, Chinese medicine index of kidney hypertrophy in rats and significantly higher compared to normal group (P <0.01) , and the model group and the two treatment groups no significant difference (P> 0.05).3 The rats fasting blood glucose (FPG) ComparisonModel group, FPG was significantly higher than the normal group, the difference was significant (P<0.01), after intervention by the administration, the two treatment group compared with the model group had no significant difference (P>0.05). Benazepril group compared with the traditional Chinese medicine was no significant difference (P> 0.05).4 The group of rats serum albumin, 24 hour urine protein quantitative and urinary beta 2 microglobulin comparisonModel groups and traditional Chinese medicine treatment in rat plasma albumin significantly lower than normal groups, Model groups and traditional Chinese medicine treatment in rat plasma albumin value than the model of two treatment groups group, P<0.05, no significant differences between the two groups of serum albumin significantly lower than normal group. Model of rat urine protein quantitative than normal groups, After two drugs in the treatment of urine protein quantitative dropped significantly, Two group model rat urine protein quantitative significantly lower than the group, P<0.01, but Chinese traditional medicine treatment groups should be higher than that of benazepril group; Urinary beta 2 microglobulin and the results of urine protein quantitative Basic Agreement.5 The serum fructosamine (FMN) ComparisonModel group, Benazepril Group, the Chinese Medicines Board of the serum fructosamine significantly higher than the normal group (P<0.01), and the model group, Benazepril group between Chinese no significant difference (P>0.05).6 Lipids of rats comparedModel of each group serum total cholesterol (TC) was significantly higher than normal group (P<0.01), traditional Chinese medicine group was significantly lower than the model (P<0.01), while Benazepril between group and model group, no significant differences (P>0.05).For triglycerides (TG), the two treatment groups were lower than model group (P<0.01), while the two treatment group compared with the normal group no significant difference (P>0.05).7 Each group comparison of renal function in ratsCompared with normal group, model group, creatinine (Scr) significantly increased, the difference was significant (p<0.05). The treatment group compared with the model group decreased significantly, there are significant differences (p<0.05). Benazepril,Chinese medicine group and no significant difference between groups (p>0.05).For blood urea nitrogen (BUN), two-treatment group and normal group were also significantly lower than model group (p <0.01), compared with the normal group, and the Chinese Medicines Board Benazepril have increased (p<0.01 ), the increase between the level of no significant difference (p>0.05).8 Immunohistochemical observationsDiffuse distribution of AGEs in the renal tubule, the glomerular expression of basic no. Semi-quantitative analysis revealed that compared with the normal group, model group to significantly enhance the expression (P<0.01), while the expression of the two treatment groups was significantly stronger than the normal group (P<0.01), and Benazepril Chinese medicine between the treatment group no significant difference in comparison (P>0.05).9 PT-PCR analysisThe results showed: normal group, the basic non-RAGE-mRNA expression, high expression of the model group band, and compared with model group, Chinese group and the Benazepril group kidney expression of RAGE-mRNA decrease.Conclusion:1 The Yiqi Yangyin Huoxue traditional Chinese medicine can reduce urinary protein in diabetic rats, reduced serum TC, TG, improve lipid metabolism; lower serum fructosamine; lower BUN and Scr, the protection of renal function.2 The Yiqi Yangyin Huoxue traditional Chinese medicine can effectively DN inhibition of rat glomerular capillary basement membrane thickening, mesangial matrix proliferation, as well as the integration of cells and slow down the progress of renal pathology.3 The Yiqi Yangyin Huoxue traditional Chinese medicine can inhibit the renal expression of tissue AGEs.4 The Yiqi Yangyin Huoxue traditional Chinese medicine can inhibit the rat renal tissue expression of RAGE-mRNA, suggesting that traditional Chinese medicine of Yiqi Yangyin Huoxue may inhibit the emergence of AGEs, and AGEs with RAGE-mRNA between the oxidative stress response thereby inhibiting the progress of DN. |
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