中文摘要：

目的：研究钙通道拮抗剂-维拉帕米（verapamil,Ver）诱导视网膜色素上皮（retinal pigment epithelium, RPE）细胞凋亡过程中钙离子及凋亡基因caspase-3变化。方法：应用80mg/L的Ver分别作用健康人眼RPE细胞12，24及48h诱导凋亡，设立对照组。逆转录聚合酶链反应（RT-PCR）检测凋亡基因caspase-3的表达，采用Fluo-3/AM负载技术，MetaFluo4.5/coolsnapfx/IX70细胞内钙离子荧光成像系统测定每组20个RPE细胞钙荧光值，并计算RPE细胞内钙浓度（［Ca2＋］i）。结果：对照组RPE细胞Ca2＋荧光分布胞核最强，胞质次之。Ver作用12，24及48h后，细胞内［Ca2＋］i明显降低（P＜0.01）。对照组RPE细胞可见caspase-3的mRNA有少量的表达。Ver作用12h后，可见caspase-3的mRNA有较高的表达，与对照组比较，具有显著性差异（P＜0.01）。随着Ver作用时间的延长，caspase-3的mRNA表达逐渐增强，在48h时有所下降。结论：Caspase-3基因表达上调及RPE细胞内钙离子稳态失调可能在Ver诱导RPE细胞凋亡中起关键作用。

英文摘要：

AIM： To study caspase-3 gene expression and [Ca2＋]i homeostasis in verapamil （Ver）-induced human retinal pigment epithelium （RPE） cells apoptosis.METHODS： Ver 80mg/L was applied in cultured human RPE cells for 12, 24 and 48 hours to induce RPE cells apoptosis. The expression of apoptotic effector gene caspase-3 was assessed by reverse transcription polymerase chain reaction （RT-PCR）. Single cell was measured using fluorescence indicator Fura-3/AM with MetaFluo4.5/coolsnapfx/IX70 intracellular Ca2＋ fluorescence imaging system.RESULTS： High levels of expression of caspase-3 mRNA were observed in normal RPE cells and it significantly increased after co-cultured with Ver. The fluorescence in resting RPE cells was strong and distributed throughout the cells. The nucleus appeared more fluorescent than the cytoplasm. Calcium fluorescence of RPE cells attenuated after co-cultured with Ver. CONCLUSION： Up-regulation of caspase-3 gene expression and disturbance of [Ca2＋]i homeostasis might play pivotal roles in Ver-induced RPE cells apoptosis.