肾淋巴循环障碍促使单肾切除大鼠肾细胞凋亡

Renal cell apoptosis induced by renal lymph circulation disorder in uninephrectomized rats

目的探讨肾淋巴循环障碍对大鼠肾脏细胞凋亡的作用,并初步探讨该作用与Bax、Bcl-2、Caspase3表达变化的关系。方法雄性Wistar大鼠48只,随机分为模型组和假手术对照组各24只。各组大鼠分别于术后第1、7、14及28天各处死6只,留取肾组织标本提取组织蛋白、mRNA和制作石蜡切片。运用Real-time PCR、Western Blot和免疫组化检测检测Bax、Bcl-2、Caspase3在肾脏组织的表达,用肾脏细胞原位凋亡(TUNEL)的方法检测肾脏凋亡细胞。结果细胞凋亡主要发生在扩张或萎缩的肾小管;模型组的凋亡指数在第14天达到高峰(P<0.01)。模型组的Bax和caspase-3蛋白和基因表达明显增强,主要位于小管间质,尤其在远端小管。相反,Bcl-2的表达明显减弱。Bax/Bcl-2比值明显增高。结论在肾淋巴循环障碍诱导的肾脏功能和结构改变的过程中,细胞凋亡显著增加,其中Bax/Bcl-2途径发挥了积极作用。

【Objective】To investigate the influence of disorder of renallymph circulation on renal cell apoptosis and the expression of Bax/Bcl-2 and Caspase3 on rat kidney. 【Methods】Forty-eight male Wister rats were randomly divided into two groups, twenty-four rats in each group. Rats in each group were sacrifice at 7, l4, 28, 56 d after operation. Nephridial tissue were obtained for extracting protein, mRNA and paraffin section. Real-time PCR, immunostaining and western blot techniques were applied to detect the expression of Bax/Bcl-2, Caspase3 in renal tissue. Apoptotic nuclei were labeled using the terminal deoxynucleotidyl transferase dUTP nick-end-label（TUNEL） technique. 【Results】TUNEL exam showed that many positive staining cells in model group rats. Cell apoptosis mainly occurred in dilate and emarcid renal tubule. Apoptotic index elevated from day 1 in model group, peaked at day 14. Expressions of Bax and caspase-3 proteins and（or） mRNA were significantly increased in model group rats, mainly in renal tubulointerstitium. Oppositely, Bcl-2 was weakly. Increasing Bax to bcl-2 ratio were also detected in model group rats.【Conclusions】Renal lymph disorder enhanced renal cells apoptosis. Bax/Bcl-2 pathway mediates apoptosis in this disease.