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。
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 Zn^2＋,a effective capacitative calcium entry（CCE） blocker. CONCLUSION： Ca^2＋ entry through voltage-dependent ca channels increases intracytoplasm Ca^2＋, then triggers further Ca^2＋ release from endoplasmic reticulum via ryanodine receptor, and the microdomains of elevated intracytoplasm Ca^2＋ are sensed by adjacent mitochondria, which ultimately lead to cell apoptosis.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2006年第7期789-796,共8页
Chinese Journal of Clinical Pharmacology and Therapeutics
关键词
血管平滑肌细胞
凋亡
钙
兰尼定
线粒体
vascular smooth muscle cell
apoptosis
calcium release
ryanodine
mitochondria
