Tbxa2r基因敲低对嘌呤霉素氨基核苷诱导的足细胞凋亡的影响

摘要

目的:观察足细胞血栓素A2受体(Tbxa2r)基因敲低对嘌呤霉素氨基核苷(puromycin aminonucleoside,PAN)诱导的足细胞凋亡的影响.方法:通过目标序列设计4条siRNA,评价细胞转染效率后,RT-PCR检测siRNA干扰效果,蛋白质印迹法检测Tbxa2r蛋白表达,流式细胞仪检测PAN诱导的MPC5足细胞的凋亡情况.结果:目标序列在转染24 h后几乎没有干扰效果,而在48h时第3条siRNA(Tbxa2r-mus-1677)干扰序列作用效果最好;在干扰72h后,Tbxa2r蛋白表达量明显降低.PAN刺激48h后,空白组与阴性对照组MPC5足细胞没有明显的凋亡现象,阳性对照组与Tbxa2r基因敲低PAN组细胞凋亡和坏死现象明显.结论:PAN能促进Tbxa2r基因敲低的足细胞发生凋亡和坏死.%Objective:To observe the effect of podocytes Tbxa2r gene knockdown on podocytes apoptosis induced by puromycin aminonucleoside (PAN).Methods:The interference effect was detected by RTPCR after designing the target sequence and evaluating the cell transfection efficiency.The expression of Tbxa2r protein was measured by western blotting.The apoptosis of MPC5 cells induced by PAN was tested by flow cytometry.Results:Target sequence after 24 h transfection almost had no interference effects,and after 48 h,the third siRNA (Tbxa2r-mus-1677) had the best interference effect ; after 72 h transfection,Tbxa2r protein expression was significantly reduced.PAN stimulated for 48 h,no significant MPC5 podocyte apoptosis were observed in the control group and negative control group,while there were significant apoptosis and necrosis in the positive control group and Tbxa2r gene knockdown PAN group.Conclusion:The PAN might facilitate the apoptosis and necrosis in Tbxa2r gene knockdown podocytes.