摘要目的 观察光损伤后人视网膜色素上皮(RPE)细胞血管内皮生长因子A(VEGF-A)及其受体可溶性fm样酪氨酸激酶受体(sFlt-1)和包含激酶植入区域受体(KDR)的表达.方法 体外培养人RPE细胞,取第8～12代生长良好的传代细胞用于实验.将同代细胞随机分为对照组和光损伤组.以18 W冷白色荧光灯作为光源,自制光照器.采用双层高压消毒锡纸包裹对照组细胞,与光损伤组细胞一同置于自制光照器下,控制光照强度在(2200±300) Lux,持续光照12 h建立光损伤模型.于光损伤后0、6、12、24 h终止培养,采用逆转录聚合酶链反应(RT-PCR)和蛋白免疫印迹法(Western blot)检测VEGF-A、sFlt-1及KDR的mRNA及蛋白表达.结果 光损伤组VEGF-A mRNA及蛋白表达在光损伤后6h时明显增加,12h时达到高峰,明显高于对照组(t=2.74,2.93;P＜0.05),随后降低.sFlt-1的mRNA及蛋白表达在光损伤后12h达到高峰,明显高于对照组(t=4.32,P＜0.01)；24 h时明显低于对照组(t=2.41,P＜0.05).KDR的mRNA及蛋白表达在光损伤后6、12 h时较对照组无明显变化(t=1.84,P＞0.05),24 h时高于对照组(t=2.89,P＜0.05).结论 光损伤后6、12h时,RPE细胞VEGF-A及sFlt-1的表达明显增高,KDR的表达相对稳定.光损伤后24 h时,RPE细胞VEGF-A及sFlt-1的表达降低,KDR的表达增高.

abstractsObjective To observe the expression of vascular endothelial growth factor A (VEGF-A) and its receptors sFlt-1,kinase insert domain receptor (KDR) in light-injured human retinal pigment epithelial (RPE) cells.Methods Cultured human RPE cells (8th - 12th generations) were divided into normal control group and light damage group.The cells of two groups were exposed to the 18 W cold white light (2200±300) Lux for 12 hours to induce light damage responses,but the cells of normal control group were packed by tinfoil with double-deck high pressure disinfection.The VEGF-A,sFlt-1 and KDR mRNA and protein expressions were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot at 0,6,12,24 hours after light damage. Results The VEGF-A mRNA and protein expressions in light damage group were significantly increased at 6 hours,and reached its peak at 12 hours after light damage which obviously higher than that in normal group (t=2.74,2.93; P＜0.05),and then went down gradually.The sFlt-1 mRNA and protein expressions in light damage group reached its peak at 12 hours after light damage which obviously higher than that in normal group (t=4.32,P＜0.01),but obviously lower than that in normal group at 24 hours after light damage (t=2.41,P＜0.05).The KDR mRNA and protein expressions in light damage group were obviously higher than that in normal group at 24hours after light damage (t=2.89,P＜0.05),but there was no changes at 6,12 hours after light damage (t=l.84,P＞0.05).Conclusions At 6,12 hours after light damage,the expressions of VEGF-A and sFlt-1 increases significantly and KDR expression is stable in light-injured RPE cells.At 24 hours after light damage,the expression of VEGF-A and sFlt-1 decreases,but KDR expression increases in light-injured RPE cells.