补体C5b-9复合物对人视网膜色素上皮细胞VEGF表达的影响

Effect of Complement 5b-9 Complex on VEGF Expression in Cultured Human RPE Cells

目的:观察补体C5b-9复合物对人视网膜色素上皮(RPE)细胞血管内皮生长因子(VEGF)表达的影响。方法:采用大鼠血清接触兔红细胞提取补体C5b-9复合物,提纯、鉴定后分别作用体外培养的人RPE细胞。光学显微镜下观察不同浓度补体C5b-9复合物对细胞的作用效果选取实验剂量。半定量RT-PCR法测量RPE细胞VEGF表达的变化。结果:补体C5b-9复合物刺激浓度>20mg/L,光学显微镜示REP细胞结构破坏;该浓度≤20mg/L,光学显微镜未见REP细胞结构明显改变,选取20mg/L补体C5b-9复合物刺激RPE细胞4h,VEGFmRNA表达水平显著增高(P<0.01)。结论:补体C5b-9复合物可诱导人RPE细胞VEGF表达上调,可能是补体C5b-9复合物参与脉络膜新生血管(CNV)形成的机制之一。

Objective： To observe the effect of complement 5b-9 （C5b-9）complex on VEGF in human cultured RPE cells. Methods： C5b-9 complex was deposited on rabbit erythrocyte by incubating rat fresh sera and rabbit erythrocyte together. The extract C5b-9 complex was purified,quantified and identification. Human RPE cells were isolated and exposed in the extracted C5b-9 complex. The amount of VEGF in cuhured RPE ceils was determined by RT-PCR. Results： RPE cells were destroyed exposed in 80 mg/L and 40 mg/L C5b-9 complex. The structure of RPE cells was not be changed obviously exposed in 20 mg/L or less C5b-9 complex.The expression of VEGF mRNA in RPE cells was higher than that in control cells after exposed in 20 mg/L C5b-9 complex for 4 hours （P 〈 0.01 ）. Conclusion： The expression of VEGF in cultured RPE cells is significantly up-regulated by sublytic C5b-9 complex, and it may be pathogenesis of CNV.