补体C3a促进人视网膜色素上皮细胞分泌血管内皮生长因子的研究  

C3a Stimulates the Secretion of VEGF in Human Retinal Pigment Epithelial Cells

作　　者：阳雪 曹晓光[2] 龙琴 

机构地区：[1]中国医学科学院北京协和医院眼科,北京100073 [2]北京大学人民医院眼科,北京100044

出　　处：《中国医科大学学报》2015年第11期966-969,974,共5页Journal of China Medical University

基　　金：国家自然科学基金(81070755)

摘　　要：目的观察补体活化产物C3a对体外培养的人视网膜色素上皮细胞(RPE)分泌血管内皮生长因子(VEGF)的影响。方法采用酶联免疫吸附测定法(ELISA)和半定量逆转录聚合酶链式反应(RT-PCR)的方法,测定浓度0.1μmol/L和0.3μmol/L的重组人C3a干预RPE细胞24、48、72 h后VEGF蛋白和mRNA表达水平。结果 0.1μmol/L的C3a干预RPE细胞72 h后,VEGF蛋白表达高于干预48 h和24 h的水平,差异均有统计学意义(P<0.05);0.3μmol/L的C3a作用RPE细胞24、48、72 h后,VEGF蛋白表达组间差异均有统计学意义(P<0.01),具有正性时间—效应关系;0.1μmol/L的C3a作用RPE细胞72 h后,VEGF mRNA表达高于48 h和24 h,差异有统计学意义(P<0.05);0.3μmol/L的C3a作用RPE细胞72 h后,VEGF mRNA表达高于48 h和24 h,差异有统计学意义(P<0.01);0.3μmol/L的C3a作用72 h时VEGF的蛋白表达量高于0.1μmol/L的C3a,差异有统计学意义(P<0.01);0.3μmol/L的C3a作用24 h时VEGF mRNA表达量高于0.1μmol/L的C3a,差异有统计学意义(P<0.01)。结论 C3a可促进人RPE细胞VEGF的分泌,高浓度C3a对VEGF分泌的促进作用早于低浓度C3a,提示补体活化程度越强,对新生血管的促进作用则越迅速。Objective To investigate the effect of C3a on vascular endothelial growth factor （VEGF） secretion in cultured human retinal pigment epithelial cells （RPECs）. Methods RPECs were cultured in the presence of C3a at different concentrations （0.1 μmol/L and 0.3 μmol/L） for 24 h, 48 h and 72 h. Enzyme linked immunosorbent assay （ELISA） and semiquantitative reverse transcription -polymerase chain reaction （RT-PCR） were used to detect the protein and mRNA levels of VEGF. Results The protein expression of VEGF influenced by C3a at the concentration of 0.1 μmol/L at the time of 72 h was higher than that of 48 h and 24 h （P 〈 0.05 ）. A positive correlation was indicated between the protein expression of VEGF influenced by C3a at the concentration of 0.3 μmol/L and the duration of this influence （P 〈 0.01 ）. The mRNA expression of VEGF influ- enced by C3a at the concentration ofO.1 μmol/L and 0.3 M, mol/L at the time of 72 h was higher than that of 48 h and 24 h （P 〈 0.05）, respectively. The protein expression of VEGF influenced by C3a at the concentration of 0.3 μmol/L was higher than that of 0.1 μmol/L at the time of 72 h （P 〈 0.01 ）. The mRNA expression of VEGF influenced by C3a at the concentration of 0.3 μmol/L was higher than that of 0.1 μmol/L at the time of 24 h （P 〈 0.01 ）. Conclusion C3a promotes the secretion of VEGF in RPECs in a concentration dependent manner, which indicates that the stronger the activation of complement is, the faster the promotion of neovascularization.

关 键 词：人视网膜色素上皮细胞 补体C3a 血管内皮生长因子 

分 类 号：R774.1[医药卫生—眼科]