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目的探讨维拉帕米(verapamil,Ver)对体外培养的人视网膜色素上皮(retinal pigment epithelium,RPE)细胞的凋亡作用,观察凋亡过程中原癌基因蛋白质(Bcl2)和碱性成纤维细胞生长因子(baic fibroblast growth factor,bFGF)的表达变化。方法应用80mg·L-1Ver作用于体外培养的人RPE细胞12h、24h、48h,吖啶橙(acridineorange,AO)荧光染色及透射电镜观察RPE细胞凋亡;逆转录聚合酶链反应(RTPCR)检测Bcl2mRNA和bFGFmRNA表达变化。结果经Ver作用的体外培养的人RPE细胞具有典型的凋亡形态学特征:核染色质浓染、边集;细胞体积缩小。同时RPE细胞Bcl2mRNA表达下降;而bFGFmRNA表达随着作用时间延长,呈增高趋势。结论Ver可诱导体外培养人RPE细胞凋亡,凋亡过程中Bcl2表达逐渐下降,而bFGF表达增强。
Objective To investigate the apoptosis of human retinal pigment epithelium (RPE) cells cultured in vitro by verapamil (Ver) and to observe the growth of proto-oncogene protein (Bcl2) and basic fibroblast (BFGF) expression changes. Methods Human RPE cells cultured in vitro were treated with 80 mg · L -1 Ve for 12 h, then stained with acridine orange (AO) for 24 h, 48 h, respectively. The apoptosis of RPE cells was observed by transmission electron microscopy. The expression of RTPCR Bcl2 mRNA and bFGF mRNA expression changes. Results The cultured human RPE cells treated with Ver had typical morphological features of apoptosis: nuclear chromatin staining, edge set, and cell volume reduction. At the same time, the expression of Bcl2 mRNA in RPE cells decreased, while the expression of bFGF mRNA increased with the extension of time. Conclusion Ver can induce the apoptosis of human RPE cells in vitro. The expression of Bcl-2 and Bcl-2 in apoptosis decreased gradually, while the expression of bFGF increased.