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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 aldosterone-induced autophagy in rat mesangial cells. Methods Rat mesangial cells were cultured and divided into aldosterone group (A group), bufalin group (B group), aldosterone + bufalin group (C group) and normal control group (D group). The morphological changes of mesangial cells were observed under inverted microscope. The autophagy of mesangial cells was observed under transmission electron microscope. Western blot was used to detect the protein expression of autophagy marker protein microtubule-associated protein 3 (LC3) and p62. Results The morphology of mesangial cells in groups A, B and C had no significant changes compared with those in group D. Compared with group D, the numbers of autophagosomes in group A increased and the expressions of LC3 and p62 were up-regulated (P <0.05). However, group C could reverse the change of these indexes in group A (P < 0.05). Conclusions Bufalin may partially relieve aldosterone-induced increase of autophagy in mesangial cells, which may be one of the mechanisms of its protective effect on kidney.