祛风通络方对糖尿病肾脏病模型大鼠足细胞损伤的影响

Effects of Qufeng Tongluo Recipe( 祛风通络方) on Podocyte Injury in Diabetic Nephropathy Model Rats

目的探讨祛风通络方治疗糖尿病肾脏病的可能作用机制。方法 32只GK大鼠随机分为模型组、假手术组、益气活血组、祛风通络组,每组8只。除假手术组外其余各组大鼠采用高脂高热量喂养加单肾切除术复制糖尿病肾脏病模型。益气活血组给予浓度为0. 75 g/ml益气活血颗粒灌胃,祛风通络组给予浓度0. 9 g/ml祛风通络颗粒灌胃,模型组和假手术组给予纯净水灌胃,剂量均为1 ml/100 g,共8周。观察大鼠一般情况,检测24小时尿蛋白变化以及血糖、肌酐、尿素氮、丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)水平,HE染色观察肾脏病理情况,采用免疫组化法检测肾组织中nephrin、desmin、podocin、podocalyxin等足细胞标志蛋白表达情况。结果 HE染色结果显示,模型组可见到肾小球肥大,少量肾小球萎缩,可见肾小管扩张;益气活血组和祛风通络均未见肾小球萎缩,系膜细胞增生较轻,肾小管上皮细胞变性较轻。模型组大鼠肌酐明显高于假手术组(P <0. 05);益气活血组大鼠ALT显著低于模型组以及祛风通络组(P <0. 05)。与模型组比较,益气活血组以及祛风通络组均可降低24小时尿蛋白(P <0. 01)。益气活血组以及祛风通络组大鼠肾组织nephrin表达高于模型组(P <0. 05或P <0. 01),祛风通络组大鼠肾组织podocin表达要高于模型组以及益气活血组,祛风通络组大鼠肾组织podocalyxin表达要高于模型组(P <0. 05)。结论祛风通络方可以缓解足细胞损伤,可能是其治疗糖尿病肾脏病的机制之一。

Objective To explore the possible mechanism of Qufeng Tongluo Recipe( 祛风通络方) in the treatment of diabetic nephropathy. Methods A total of 32 GK rats were randomly divided into model group,sham operation group,Yiqi Huoxue group and Qufeng Tongluo group,with 8 rats in each group. Except the sham operation group,rats were treated with high-fat,high-calorie feeding plus single nephrectomy to replicate the diabetic kidney disease model. Yiqi Huoxue group was given Yiqi Huoxue Granules at a concentration of 0. 75 g/ml by gavage,and the Qufeng Tongluo group was given a concentration of 0. 9 g/ml Qufeng Tongluo Granules by gavage. The model group and the sham operation group were given pure water by gavage. The dose of all groups was 1 ml/100 g for 8 weeks.The general condition of rats was observed,and the 24-hour urine protein changes,levels of blood glucose,creatinine,urea nitrogen,alanine aminotransferase( ALT),and aspartate aminotransferase( AST) were detected. HE staining was used to observe renal pathology. The expression of podocyte marker proteins of nephrin,desmin,podocin and podocalyxin was detected by immunity histochemistry. Results The results of HE staining showed that the model group showed glomerular hypertrophy,a small amount of glomerular atrophy,and renal tubular dilatation.There was no glomerular atrophy in the Yiqi Huoxue group and Qufeng Tongluo group,and the mesangial cells proliferated lightly. Renal tubular epithelial cells were less degenerative. The creatinine of the model group was significantly higher than that of the sham operation group( P < 0. 05). The ALT of the Yiqi Huoxue group was significantly lower than that of the model group and the Qufeng Tongluo group( P < 0. 05). Compared with the model group,Yiqi Huoxue group and Qufeng Tongluo group could reduce 24-hour urine protein( P < 0. 01). The expression of nephrin in the kidney of Yiqi Huoxue group and Qufeng Tongluo group was higher than that in the model group( P < 0. 05 or P < 0. 01). The expression of podocin in the kidney of the rats in the Qufeng Tongluo group was higher than that in the model group and Yiqi Huoxue group. The expression of podocalyxin in the kidney tissue of the rats in the Qufeng Tongluo group was higher than that in the model group( P < 0. 05). Conclusion Qufeng Tongluo Recipe can alleviate podocyte injury and that may be one of the mechanisms for its treatment of diabetic nephropathy.