自噬在膜性肾病补体攻膜复合物所致足细胞损伤中的作用

Effect of autophagy on podocyte injury induced by complement membrane attack complex in membranous nephropathy

目的探讨自噬在补体攻膜复合物C5b-9所致足细胞损伤中的作用。方法利用酵母多糖活化正常人血清制作攻膜复合物C5b-9;分别用含C5b-9的酵母多糖活化血清(ZAS)、雷帕霉素、氯喹处理MCP-5足细胞,透射电子显微镜观察自噬泡形成,蛋白印迹、免疫荧光检测LC3、F-actin、podocin、nephrin表达。结果在ZAS刺激下,足细胞内LC3Ⅱ表达明显增加,自噬泡生成增多,骨架结构严重破坏。氯喹处理足细胞加重骨架结构破坏、下调F-actin、podocin和nephrin表达,而雷帕霉素减轻上述损伤(P<0.01或0.05)。结论自噬可保护足细胞抵抗亚溶解型C5b-9所致损伤。

Objective To explore the role of autophagy in podocyte injury induced by complement membrane attack complex C5b-9.Methods C5b-9 was prepared by coculture of normal human serum and yeast polysaccharide.MPC-5 podocytes were treated with zymosan activation serum(ZAS)containing C5b-9,rapamycin or chloroquine.The autophagosomes were observed by transmission electron microscopy,while LC3,F-actin,podocin and nephrin expression was detected by Western blot and immunofluorescence.Results ZAS treatment upregulated LC3 II expression and autophagosome formation,and destroyed skeletal structure in podocytes.Co-treatment with chloroquine aggravated the cytoskeleton damage and downregulated expression of F-actin,podocin and nephrin,while rapamycin alleviated ZAS-induced injuries in podocytes(P<0.01 or 0.05).Conclusion Autophagy can protect podocytes from sublytic C5b-9 induced injury.