益肾化湿颗粒对糖尿病肾病小鼠中Yes相关蛋白的调控作用

Regulation of Yes associated protein in diabetic nephropathy mice by Yishen Huashi Granule

目的探讨益肾化湿颗粒(Yishen Huashi Granule,YSHS)对链脲佐菌素(STZ)诱导的糖尿病肾病(DN)小鼠的保护作用及可能的相关机制。方法建立STZ诱导DN小鼠模型,随机分为模型组、YSHS组及YAP抑制剂维替泊芬组,同时设立对照组。造模成功后8周开始干预,疗程4周。分别检测各组干预前后尿蛋白及干预后血肌酐、尿素氮水平。疗程结束后处死小鼠,行光镜HE染色观察肾小球病理变化,Western blot检测YAP、p-YAP S127及CTGF的蛋白表达,RT-PCR检测YAP、CTGF及足细胞蛋白nephrin、podocalyxin、WT1的mRNA表达。结果YSHS组在生化指标上较模型组均有好转,HE染色显示肾小球病理损伤有所改善。模型组YAP、p-YAP S127及CTGF的蛋白表达增多,YAP和CTGF的mRNA表达亦增多,而3种足细胞蛋白的mRNA表达减少。经YSHS治疗后,YAP、p-YAP S127及CTGF的蛋白表达与YAP、CTGF的mRNA表达量均有所下降,而nephrin、podocalyxin、WT1的mRNA表达量升高。结论YSHS可降低DN小鼠尿蛋白、保护肾功能并减轻肾小球病理损伤,其机制可能与下调肾组织中YAP,减少CTGF表达水平并上调足细胞蛋白的mRNA表达有关。

Aim To explore the protective effect of Yishen Huashi Granule(YSHS)on streptozotocin(STZ)-induced diabetes nephropathy(DN)in mice and its possible mechanism.Methods The STZ induced DN mice model was established,which was randomly divided into model group,YSHS group and YAP inhibitor Vertepofin group,and the control group was also established.The intervention was started eight weeks after the successful modeling with the course of four weeks.Urine protein concentration before and after intervention in each group as well as serum creatinine(Scr)and blood urea nitrogen(Bun)levels after intervention were measured.After the treatment,the mice were sacrificed,and the pathological changes of glomeruli were observed by light microscope HE staining.The protein expression of YAP,p-YAP S127 and CTGF were detected by Western blot,and the mRNA expressions of YAP,CTGF and podocyte functional proteins nephrin,podophyxin and WT1 were detected by RT-PCR.Results The biochemical indexes of YSHS group were better than those of model group,and HE staining showed that the pathological injury of glomeruli was improved.In the model group the protein expression of YAP,p-YAP(S127)and CTGF as well as the mRNA expression of YAP and CTGF increased,while the mRNA expression of nephrin,podocalyxin and WT1 decreased.After YSHS treatment,the protein expression of YAP,p-YAP S127,CTGF and the mRNA expression of YAP and CTGF decreased,while the mRNA expression of nephrin,podocalyxin and WT1 increased.Conclusions YSHS can reduce urinary protein,protect renal function and alleviate glomerular pathological injury in DN mice.Its possible mechanism may be related to the down-regulation of YAP in renal tissue,the reduction of CTGF expression level and the up-regulation of podocyte protein mRNA expression.