锌离子对脂多糖作用下大鼠腹膜间皮细胞凋亡的影响及机制  

The effect of zinc ion on LPS-induced cell apoptosis in rat peritoneal mesothelial cells and its mechanism

作　　者：张秀丽[1,2] 鲁淑敏 赵越[2] 马健飞[3] 李德天[4] 李红娟[5] 

机构地区：[1]本溪市中心医院肾内科 [2]中国医科大学细胞生物国家重点实验室 [3]中国医科大学第一附属医院肾内科 [4]中国医科大学第二附属医院肾内科 [5]中国人民解放军95935部队

出　　处：《中国血液净化》2014年第4期313-316,共4页Chinese Journal of Blood Purification

摘　　要：目的研究锌离子对脂多糖(LPS)诱导的大鼠腹膜间皮细胞(RPMC)凋亡的影响及机制的探索,从而为锌离子在腹膜透析中的应用提供实验基础。方法胰蛋白酶消化法培养原代大鼠腹膜间皮细胞、传代、经鉴定后分组:①对照组;②LPS组;③ZnSO4组;④ZnSO4/LPS组。应用AnnexinV-FITC凋亡检测试剂盒及流式细胞仪检测RPMC凋亡率,并采用Westen Blot方法检测细胞外信号调节激酶(PERK),BAX,凋亡诱导因子(AIF),天冬氨酸特异性半胧氨酸蛋白酶3(caspase3),天冬氨酸特异性半胧氨酸蛋白酶9(caspase9)蛋白表达。应用流式细胞术,采用活性氧自由基(ROS)检测试剂盒检测各组别的ROS蛋白表达情况。结果与对照组相比,LPS组RPMC凋亡细胞数量和相关凋亡蛋白(BAX,AIF,caspase3,caspase9)表达升高(P<0.01)。与LPS组相比,ZnSO4作用组RPMC凋亡细胞数量和相关凋亡蛋白表达明显降低(P<0.01)。LPS组ROS显著高于对照组而ZnSO4作用组ROS显著低于LPS组(P<0.01,P<0.01)。与LPS组相比,ZnSO4作用组P-ERK的表达明显升高(P<0.01)。结论 ZnSO4可能可以逆转LPS所致的RPMC凋亡,对腹膜透析相关腹膜炎过程中所导致的RPMC损伤具有保护作用,其作用机制可能是通过ERK通路而发挥作用。Objective To explore whether Zn＋2 can be used in peritoneal dialysis, we investigated the effect of Zn＋ 2 on lipopolysaccharide （LPS）-induced cell apoptosis in rat peritoneal mesothelial cells （RPMCs） and its underlying mechanism. Methods RPMCs were isolated by enzyme disaggregation, cul- tured, and then identified by phase contrast microscopy, transmission electron microscopy, and immunocyto- chemistry methods. RPMCs were incubated with 100 μg/ml LPS for 24 hours, or stimulated by 100 μg/ml LPS after incubation with 50 μM ZnSO4 for 24 hours. RPMCs incubated in regular medium were used as the controls. The expressions ofp-ERK, BAX, AIF, caspase 3, and caspase 9 were assayed by western blot. Reac- tive oxygen species was measured by a reactive oxygen species assay kit. Results ZnSO4 significantly inhib- ited the LPS-induced RPMC apoptosis by attenuating ROS production, inhibiting LPS-induced BAX, AIF, and the activation of caspase 3 and caspase 9. Further studies revealed that ZnSO4 treatment facilitated cell sur- vival through the activation of ERK signaling pathway. Conclusion These results indicate that Zn＋2 inhib- its LPS-induced apoptosis in RPMCs by attenuating ROS production and inhibiting BAX, AIF, and the activa- tion of caspase 3 and caspase 9. The ERK signaling pathway may take part in these changes.

关 键 词：脂多糖 ZnSO4 凋亡 大鼠腹膜间皮细胞 腹膜透析 

分 类 号：R459.5[医药卫生—治疗学]