造影剂诱导肾小球内皮细胞凋亡的caspase-3机制

The role of caspase-3 in the apoptosis of glomerular endothelial cells induced by contrast media

目的研究半胱氨酸蛋白水解酶caspase-3在非离子型低渗造影剂诱导肾小球内皮细胞凋亡中的作用,探讨非离子型低渗造影剂诱导肾小球内皮细胞凋亡的可能机制。方法DNA凝胶电泳分析不同浓度的非离子型低渗造影剂碘佛醇(ioversol)诱导肾小球内皮细胞凋亡情况,Westernblot分析caspase-3活化片段蛋白的表达变化,AnnexinV荧光染色和流式细胞检测caspase-3特异性阻断剂对细胞凋亡的抑制作用。结果高浓度的碘佛醇作用24h后,DNA凝胶电泳呈现典型的凋亡梯状条带,肾小球内皮细胞caspase-3活化片段蛋白表达明显升高,其升高幅度与碘佛醇浓度密切相关;AnnexinV荧光标记染色发现,caspase-3特异性阻断剂可以明显减轻碘佛醇诱导的肾小球内皮细胞凋亡。结论非离子型低渗造影剂诱导肾小球内皮细胞凋亡与caspase-3的活化有关,肾小球内皮细胞凋亡可能参与了造影剂肾病的发生。

Aim To investigate the role of caspase-3 in glomerular endothelial cells apoptosis induced by nonionic low osmolar contrast media （LOCM）. Methods After being treated with ioversol at different dosages, the apoptosis of glomerular endothelial cells was analyzed by DNA electrophoresis, and the protein level of caspase-3 cleaved fragments was analyzed by Western blotting. The annexin V stain and flow cytometry assay were applied to explore the effect of caspase-3 inhibitor on the amelioration of glomerular endothelial cells apoptosis. Results Following treatment with high concentration of ioversol for 24 h, a typical DNA ladder determined by agarose gel electrophoresis was observed. The protein level of caspase-3 cleaved fragments was increased obviously and related with the concentration of ioversol. The apoptotic cells positively stained with annexin V induced by ioversol was markedly reduced by caspase-3 inhibitor. Conclusion The activation of caspase-3 may be one of the main causations for the apoptosis of glomerular endothelial cells induced by non-ionic LOCM, which is contributed to the contrast media-induced nephropathy.