活血降糖胶囊对糖尿病肾病大鼠足细胞相关分子蛋白表达的影响

Influence of Huoxue Jiangtang Capsules on expression of podocyte associated proteins in diabetic nephropathy rats

目的:探讨活血降糖胶囊对糖尿病肾病(DN)大鼠足细胞相关分子蛋白Nephrin、Podocin、CD2AP表达的影响。方法:应用单侧肾脏切除结合小剂量STZ单次腹腔注射方法建造DN大鼠模型,随机分为模型组(M组)、假手术正常对照组(N组)、活血降糖胶囊干预治疗组(H组),每组6例。H组给予活血降糖胶囊的剂量折合生药2.5 g·kg^(-1)·d^(-1)灌胃,M组和N组给予等量纯化水灌胃。8周后检测各组大鼠的24 h尿蛋白定量、血生化等指标,应用免疫组化技术检测肾组织Nephrin、Podocin、CD2AP蛋白表达的情况。结果:8周后,与N组比较,M组出现24 h尿蛋白定量增多,内生肌酐清除率(Ccr)下降(P<0.05),肾组织Nephrin、Podocin、CD2AP蛋白表达水平均下调(P<0.05);与M组比较,H组24 h尿蛋白定量降低(P<0.05),Ccr升高(P<0.05),肾组织Nephrin、Podocin、CD2AP蛋白表达水平均上调(P<0.05)。采用直线相关分析后发现,24h尿蛋白定量与肾组织Nephrin、Podocin、CD2AP蛋白表达水平呈负相关;Ccr与肾组织Nephrin、Podocin、CD2AP蛋白表达水平呈正相关。结论:DN时存在足细胞相关分子Nephrin、Podocin、CD2AP蛋白表达异常,活血降糖胶囊对DN大鼠肾脏的保护作用机制可能是通过增加足细胞相关分子nephrin、podocin、CD2AP蛋白表达,减少蛋白尿而实现的。

OBJECTIVE To investigate influence of Huoxue Jiangtang Capsules on expression of podocyte associated proteins（Nephrin,Podocin,CD2AP）in diabetic nephropathy（DN）rats.METHODS Rats were removed one kidney and injected intraperitoneally with low dose streptozocin（STZ）to establish DN models,then divided randomly into model group（group M）,sham-operation normal control group（group N）and Huoxue Jiangtang capsule group（group H）.Every group had 6 rats.Rats received Huoxue Jiangtang capsules intraperitoneally at 2.5 g·kg^（-1）·d^（-1） in group H,and the same dose of distilled water in group M and N.The 24 hour urinary protein and blood biochemical markers were measured,expression levels of Nephrin,Podocin and CD2AP were detected by immunohistochemistry technique in every group at 8th week.RESULTS Compared with group N,24 hour urinary protein count was increased and creatine clearance ratio（Ccr）decreased（P〈0.05）,the expression of Nephrin,Podocin and CD2AP was reduced in group M（P〈0.05）.As compared with group M,the 24 hours urinary protein count was decreased（P〈0.05）and Ccr increased（P〈0.05）,expression levels of Nephrin,Podocin and CD2AP were increased in group H（P〈0.05）.The 24 hours urinary protein count was negatively correlated with expression of Nephrin,Podocin and CD2AP,while Ccr was positively correlated with expression of Nephrin,Podocin and CD2AP.CONCLUSIONExpression levels of Nephrin,Podocin and CD2AP are abnormal in DN.Protective effects of Huoxue Jiangtang Capsules on kidney in DN rats are achieved by upregulating expression levels of podocyte-associated proteins（Nephrin,Podocin,CD2AP）and reducing 24 hour urinary protein count.