蟾蜍灵对脂多糖诱导的大鼠肾小球系膜细胞凋亡的影响

Effect of bufalin on apoptosis of rat glomerular mesangial cells induced by lipopolysaccharide

目的:探讨蟾蜍灵(bufalin)对脂多糖(lipopolysaccharides,LPS)诱导的大鼠肾小球系膜细胞(glomerular mesangial cell,GMC)凋亡的影响。方法:体外培养大鼠肾小球系膜细胞,分为对照组、LPS刺激组和蟾蜍灵干预组,应用台盼蓝拒染法检测蟾蜍灵对GMC的细胞毒性作用,应用透射电镜观察系膜细胞超微结构的变化,采用逆转录PCR检测Bax和Bcl-2基因mRNA的表达,Western blot法检测Bax和Bcl-2的蛋白表达。结果:透射电镜观察蟾蜍灵干预组可见凋亡早期形态学改变。逆转录PCR和Western blot结果显示蟾蜍灵组较脂多糖组Bax表达增多,Bcl-2表达减少,差异均具有统计学意义(P<0.01),蟾蜍灵干预后,Bax较Bcl-2表达过量。结论:蟾蜍灵可能通过调节Bax和Bcl-2的相对表达量而诱导脂多糖作用后肾小球系膜细胞发生凋亡。

Objective：To investigate the effects of bufalin on glomerular mesangial cells （GMC）apoptosis induced by lipopolysaccharide （LPS）. Methods： Rat GMC lines were cultivated in vitro and divided into 3 groups：control group, LPS group （LPS 5mg/L）, bufalin group（LPS 5 mg/L and bufalin 1×10^-8 mol/L）.Cell structure was observed by electron microscrop.The transcriptional level of Bax and Bcl-2 gene were detected by RT-PCR. The protein expression level of Bax and Bcl-2 were detected by Western blot method. Results： The morphology of GMC presented changes of apoptosis with electron microscopg. The mRNA and protein expression of Bax in bufalin group were more significant than those in LPS group （P 〈 0.01）, but the mRNA and protein expression of Bcl-2 were less than those in LPS group（P 〈 0.01）. Conclusion：Bufalin may induce the cell apoptosis of lipopolysaccharide-induced GMC by modulating Bax and Bcl-2.