Objective:To assess the influnce of triptolide on the apoptosis of podocytes in adriamycin nephropathy (ADR) mice and to explore potential mechanisms.Methods:Methods Wistar clear mice were randomly divided into three groups:normal control group(N group),adriamycin-induced nephropathy group(ADR group) and triptolide-treated ADRN group (T group).The ADR group and T group were injected with adriamycin(6.5 mg/kg) via tail-vein for one time.Twentity four-hour urinary protein was checked at the end of 2,4,6,8weeks respectively.Then we measured the biochemical parameters,observed the pathological changes of the kidney,counted the apoptosis of pcodcytes of mice using TUNEL method.The number of podocyte were determined by WT-1 immunohistochemistry,the mRNA levels of Synaptopodin,p53,Notch 1 were determined by RT-PCR.The expression of Notch 1,HES 1 were determined by Western blotting.Results:(1).Compared with the N group,the level of 24h urinary protein,BUN,SCr,CysC of ADR mice were significantly increased (P ＜ 0.05).The number of podocytes were significantly reduced (P ＜ 0.05) with wider,mesangial region and mesangial hyperplasia.In contrast 24h urinary protein,SCr,CysC were decreased in T group and podocyte number were increased(P ＜0.05),and the pathological changes of the kidney were improved.(2).Synaptopodin mRNA and protein expression were markedly decreased,but p53 mRNA expression was increased in ADR group (P ＜ 0.05),with increased volume of podocytes and podocytes than N group (P ＜ 0.05).However,synaptopodin mRNA and protein expression were increased,and p53 mRNA expression was decreased with less apoptotic podocytes in T group that ADR group (P ＜ 0.05).(3).Compared with N group,the mRNA and protein expression of Notch 1 and HES 1 were higher in ADR group(P ＜ 0.05).The expression of Notch 1 and HES 1 were decreased in T group (P ＜ 0.05).Conclusion:Triptolide may ameliorate the ADR-induced podocyte apotosis through inhibiting Notch signaling pathway.%目的:观察雷公藤甲素对阿霉素肾病(ADRN)大鼠足细胞凋亡的影响并初步其可能机制.方法:将45只Wistar大鼠随机分为对照组(N)、模型组(ADR)及治疗组(T).其中ADR组和T组给予一次性尾静脉注射阿霉素6.5 mg/kg构建ADRN模型大鼠,T组给予雷公藤甲素(200 μg·kg-1·d-1)干预8周.于2、4、6、8周分别检测24 h尿蛋白定量,观察各组大鼠的生化指标及肾组织学改变,TUNEL法测足细胞凋亡,WT-1免疫组织化学染色检测肾小球足细胞数,RT-PCR检测Synaptopodin、p53、Notch1 mRNA表达变化,western印迹测定Synaptopodin、Notch1、Hes1的蛋白表达.结果:(1)ADR组各时间点24 h尿蛋白定量、尿素氮(BUN)、肌酐(Scr)胱抑素C(CysC)均高于N组,而足细胞计数显著降低,肾小球系膜区增宽,且系膜细胞增生,T组Scr、CysC低于ADR组(P＜0.05),足细胞数明显增加(P＜0.05),同时病理改变较其减轻.(2)8周末时ADR组Synaptopodin mRNA和蛋白表达降低,p53 mRNA表达升高,足细胞体积肿胀增大,凋亡足细胞数目比N组明显增多(P＜0.05);与ADR组相比,T组Synaptopodin mRNA和蛋白表达增加,p53 mRNA表达下降,凋亡足细胞数目较ADR组明显减少(P＜0.05).(3)ADR组Notch1 mRNA和蛋白的表达以及Hes1蛋白的表达均高于N组(P＜0.05),T组Notch1mRNA和蛋白的表达及Hes 1蛋白的表达低于ADR组(P＜0.05).结论:雷公藤甲素可抑制ADRN的足细胞凋亡,其保护足细胞的作用可能与降低Notch信号通路的表达有关.
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