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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。
AIM: To investigate the induction of vascular endothelial growth factor (VEGF) expression by cobalt chloride on cultured rabbit retinal pigment epithelium (RPE) cells and the effect of Genistein on its expression. Methods: Immunofluorescence histochemistry and confocal laser scanning microscope were used to express the expression of VEGF by fluorescence ratio. Semi-quantitative analysis of the induction of VEGF protein expression in RPE cells incubated with cobalt chloride at different times and the effect of gen on hypoxia Or cobalt chloride-induced VEGF expression. Results: The control group had a low level of VEGF protein expression. With the increase of hypoxia or cobalt chloride treatment time, the fluorescence ratio changed in a time-dependent manner, reaching the peak at 6 h. Compared with control group, Gen (50, 100, 200μmol / L) and Genistein (DMSO) groups both inhibited the expression of VEGF induced by cobalt chloride, and the inhibitory effect was dose-dependent. CONCLUSION: Gen inhibits VEGF expression induced by cobalt chloride in RPE cells, suggesting that Gen may have potential preventive and therapeutic value in the formation of retinal neovascularization.