钙诱导的钙释放引起一株血管平滑肌细胞株凋亡

摘要/Abstract

摘要： 目的对钙诱导的钙释放引起血管平滑肌细胞株A10 凋亡的可行性及其钙调节机制进行初步研究。方法以钾离子孵育并以多种工具药进行干预, 观测对A10 细胞细胞核形态学, DNA 琼脂糖凝胶电泳, 细胞凋亡率, 细胞线粒体膜电位的影响。结果钾离子孵育能浓度相关性地促进A10 细胞凋亡;硝本地平、ryanodine 、BAPTA AM 、环胞素对中浓度钾离子孵育所致A10 细胞凋亡有抑制作用;Zn²⁺ 、肝素对钾离子所致A10 凋亡无抑制作用。结论钾离子孵育将导致外钙经电压依赖性钙通道内流, 通过升高胞浆内钙离子浓度激活ryanodine 受体而不是IP3 受体释放内钙, 促进线粒体大量摄取而致A10细胞凋亡, 该过程可能需要内钙持续释放但与容量依赖性外钙内流无关。

关键词: 血管平滑肌细胞, 凋亡, 钙, 兰尼定, 线粒体

Abstract: AIM: To investigate the apoptosis of a vascular smooth muscle cell line A10 caused by mild K+depolarization.METHODS: Apoptosis was evaluated by nuclear staining, DNA fragmentation gel electrophoresis and propidium iodide-stained flow cytometry.Mitochondrial transmembrane potential (Δψm) was measured by flow cytometry.RESULTS: K+depolarization caused dose correlated A10 cells apoptosis; nifedipine, BAPTA AM, ryanodine inhibited the cytotoxic effect of K+completely.The combination use of nifedipine and cyclosporin A made it clear that mitochondria was involved in the apoptosis of A10 cells, and Δψm measurement further confirmed this speculation; A10 apoptosis caused by K+depolarization was not influenced by heparin or Zn²⁺, a effectivecapacitative calcium entry(CCE) blocker.CONCLUSION: Ca²⁺entry through voltage-dependent ca channels increases intracytoplasm Ca²⁺, then triggers furtherCa²⁺release from endoplasmic reticulum via ryanodine receptor, and the microdomains of elevated intracytoplasmCa²⁺are sensed by adjacent mitochondria, which ultimately lead to cell apoptosis.

Key words: vascular smooth muscle cell, apoptosis, calcium release, ryanodine, mitochondria

中图分类号:

R969

吴玉林, 马秉亮, 戴小鸣, 陈琳琳. 钙诱导的钙释放引起一株血管平滑肌细胞株凋亡[J]. 中国临床药理学与治疗学, 2006, 11(7): 789-796.

WU Yu-lin, MA Bing-liang , DAI Xiao-ming, CHEN Lin-lin. Calcium caused calcium release causes a vascular smooth muscle cell line A10 cells apoptosis[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2006, 11(7): 789-796.

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