中国血液净化

中国血液净化 ›› 2014, Vol. 13 ›› Issue (04): 313-316.doi: 10.3969/j.issn.1671-4091.2014.04.006

• 基础研究 • 上一篇    下一篇

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

张秀丽1.2,鲁淑敏1赵越2,马健飞3,李德天4,李红娟5   

  1. 117000 本溪，1本溪市中心医院肾内科
    110001 沈阳，2中国医科大学细胞生物国家重点实验室
    110001 沈阳，3中国医科大学第一附属医院肾内科
    110001 沈阳，4中国医科大学第二附属医院肾内科
    110001 沈阳，5中国人民解放军95935 部队
  • 收稿日期:2013-09-13 修回日期:2013-12-15 出版日期:2014-04-12 发布日期:2014-04-12
  • 通讯作者: 张秀丽 zhangxiuli54321@sina.com E-mail:jfeima@sohu.com

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

  • Received:2013-09-13 Revised:2013-12-15 Online:2014-04-12 Published:2014-04-12

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摘要： [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； (2) LPS组： 100μM LPS作用24h； (3) ZnSO4组：50μM ZnSO4作用24h； (4) ZnSO4/LPS组：50μM ZnSO4作用24h再加100μM LPS作用24h。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P-ERK ， BAX， 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。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。LPS组ROS显著高于对照组而ZnSO4作用组ROS显著低于LPS组。与LPS组相比，ZnSO4作用组P-ERK的表达明显升高。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。

关键词: 脂多糖, ZnSO4, 凋亡, 大鼠腹膜间皮细胞, 腹膜透析

Abstract: 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, cultured, and then identified by phase contrast microscopy, transmission electron microscopy, and immunocytochemistry 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 of p-ERK, BAX, AIF, caspase 3, and caspase 9 were assayed by western blot. Reactive oxygen species was measured by a reactive oxygen species assay kit. Results ZnSO4 significantly inhibited 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 survival through the activation of ERK signaling pathway. Conclusion These results indicate that Zn+2 inhibits LPS-induced apoptosis in RPMCs by attenuating ROS production and inhibiting BAX, AIF, and the activation of caspase 3 and caspase 9. The ERK signaling pathway may take part in these changes.

Key words: lipopolysaccharide, ZnSO4, apoptosis, rat peritoneal mesothelial cells, peritoneal dialysis