蟾蜍灵对醛固酮诱导的大鼠系膜细胞自噬的影响

Effect of bufalin on autophagy of rat mesangial cells induced by aldosterone

目的探讨蟾蜍灵对体外醛固酮诱导的大鼠系膜细胞自噬的影响。方法体外培养大鼠系膜细胞,分为醛固酮组(A组)、蟾蜍灵组(B组)、醛固酮+蟾蜍灵组(C组)和正常对照组(D组)。倒置显微镜下观察系膜细胞的形态变化,透射电子显微镜下观察系膜细胞的自噬水平,采用Western blot法检测自噬标志蛋白微管相关蛋白轻链3(LC3)和p62的蛋白表达。结果 A、B、C组系膜细胞形态与D组相比均无明显变化。与D组相比,A组细胞自噬体数量增加,LC3和p62蛋白表达上调(P<0.05);而经蟾蜍灵干预后,C组能明显逆转A组上述各指标的变化过程(P<0.05)。结论蟾蜍灵可部分缓解醛固酮诱导的系膜细胞自噬增加,这可能是其发挥肾脏保护作用的机制之一。

Objective To investigate the effect of bufalin on the autophagy of rat mesangial cells induced by aldosterone in vitro.Methods Rat mesangial cells were cultured in vitro and divided into 4groups of A（stimulated with aldosterone 10-6 mol/L）,B（stimulated with bufalin 10-8 mol/L）,C（stimulated with aldosterone 10-6 mol/L and bufalin 10-8 mol/L）and D（normal control）.The morphology and autophagy of mesangial cells were observed under inverted microscope and transmission electron microscopy,respectively.The protein expressions of light chain 3（LC3）and p62 were detected by Western blot.Results There were no significant differences in morphology of mesangial cells among four groups.Compared with group D,the number of autophagosome was increased,and the protein expressions of LC3 and p62were up-regulated in group A（P〈0.05）,which were obviously reversed by bufalin in group C（P〈0.05）.Conclusion Bufalin can partially inhibit aldosterone-induced autophagy of mesangial cells,which may be one of the underlying mechanisms for kidney protection by bufalin.