PUMA-α蛋白在高糖诱导的人腹膜间皮细胞凋亡及纤维化中的作用

摘要

Objective To explore the effects of p53 up-regulated modulator of apoptosis (PUMA)-α protein on the apoptosis and fibrosis of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods HPMCs were induced by 50mmol/L D type glucose or mannitol for 72 hours respectively,flow cytometry was employed to detect the rate of apoptotic cells,and theexpression levels of apoptosis-and fibrosis-related proteins were detected by Western blotting.The untreated HPMCs were transfected with Lenti-PUMA-α,and the treated cells were transfected with shRNA-PUMA-α,the number of apoptotic cells and the expression levels of apoptosis-and fibrosis-related proteins were detected with the methods mentioned above.Results Flow cytometry showed that the rate of apoptotic HPMCs increased after being induced by high glucose for 72 hours,and Western blotting revealed that the expression levels of pro-apoptotic and pro-fibrotic related proteins increased,but the arrestins of apoptosis and fibrosis-related proteins decreased.Up-regulation of PUMA-α promoted apoptosis and fibrosis,while down-regulation of PUMA-α alleviated apoptosis and fibrosis of HPMCs.Conclusion High glucose may accelerate apoptosis and fibrosis of HPMCs by up-regulating the expression of PUMA-α.%目的 探讨pS3上调凋亡调节因子(PUMA)-α蛋白在高糖刺激诱导的人腹膜间皮细胞(HPMC)凋亡及纤维化中的作用.方法 以50mmol/L D型葡萄糖和甘露醇分别刺激HPMC 72h后,采用流式细胞仪检测细胞凋亡率,Western blotting检测凋亡及纤维化相关蛋白的表达.分别以Lenti-PUMA-α和shRNA-PUMA-α转染正常的HPMC和高糖刺激72h后的HPMC,采用流式细胞仪检测凋亡细胞数目,Western blotting检测凋亡及纤维化相关蛋白的表达.结果 高糖刺激HPMC 72h后,流式细胞仪检测结果显示细胞凋亡率增加;Western blotting检测结果显示,促进凋亡及纤维化的蛋白表达增加,而抑制凋亡及纤维化的蛋白表达减少.上调PUMA-α的表达可促进HPMC的凋亡及纤维化;下调PUMA-α的表达可抑制HPMC的凋亡及纤维化(P＜0.05).结论 高糖刺激通过上调PUMA-α的表达,促进了HPMC的凋亡及纤维化.