金雀异黄素对氯化钴诱导的兔视网膜色素上皮细胞血管内皮生长因子表达的影响

Effects of Genistein on Cobalt Chloride-induced Vascular Endothelial Growth Factor Expression in Retinal Pigment Epithelium Cells

目的:研究氯化钴对体外培养的兔视网膜色素上皮(RPE)细胞血管内皮生长因子(VEGF)表达的诱导作用,以及金雀异黄素(genistein,Gen)对其表达的影响。方法:采用免疫荧光组织化学技术和激光共聚焦显微镜,以荧光比值来表示VEGF的表达,半定量分析氯化钴处理不同时间对体外培养的RPE细胞VEGF蛋白表达的诱导作用,以及Gen对低氧或氯化钴诱导的VEGF表达的影响。结果:对照组有低水平VEGF蛋白的表达,随着低氧或氯化钴处理时间的延长,荧光比值的变化表现为时间依赖性,在6 h点达到高峰。Gen(50、100、200μmol/L组)与二甲基亚砜(DMSO)对照组相比,均可明显抑制氯化钴诱导的VEGF的表达,且此种抑制作用呈浓度依赖性。结论:Gen可抑制氯化钴诱导的RPE细胞VEGF表达,提示Gen对视网膜新生血管的形成具有潜在的预防及治疗价值。

Objective: To investigate the upregulation of vascular endothelial growth factor (VEGF) expression by cobalt chloride (CoCl2)-simulated hypoxia on retinal pigment epithelium cells, and the effects of genistein on cobalt chloride-induced VEGF expression. Methods: RPE cells were treated with CoCl2(125 μmol/L) for different hours or pretreated with genistein. Then using im-munofluorescence and a confocal laser scanning microscope coupled to a computer, we measured fluorescence intensity ratio which correlates with the expression of VEGF on the treated cells. Results: The expression of VEGF could be upregulated by cobalt chloride-simulated hypoxia on RPE cells. The fluorescence intensity ratio increased in a time-dependent manner, peaked at 6h. Cobalt chloride-induced VEGF expression could be concentration-dependently inhibited by genistein. The inhibitory effects of genistein(50, 100,200 μmol/L group) were significant compared to dimethyl sulfoxide (DMSO; negative control group). Conclusion: Genistein could inhibite cobalt chloride-induced VEGF expression on RPE cells. It suggested that genistein might play a role in the prevention and treatment of retinal neovascularization.