康柏西普对高糖环境下人视网膜微血管内皮细胞屏障功能及VEGF表达的影响

Effects of conbercept on human retinal microvascular endothelial cells barrier function and expression of VEGF in high glucose environment

目的探讨康柏西普对高糖环境下人视网膜微血管内皮细胞屏障功能及VEGF表达的影响。方法将人视网膜血管内皮细胞分为正常组(5.5 mol/L葡萄糖)、高糖组(25 mol/L葡萄糖)、高糖+不同浓度(25μg/L、2.5μg/L、0.25μg/L)的康柏西普组。应用酶联免疫吸附测定(ELISA)检测各组细胞上清液中血管内皮生长因子(VEGF)的浓度;跨膜电阻(TER)及辣根过氧化物酶(HRP)通透性的检测,以评价细胞屏障的稳定性;蛋白质印迹法(Western Blot)检测紧密连接相关蛋白occludin的表达。结果 ELISA结果示:高糖组分泌的VEGF比正常组高,增加差异具有统计学意义(P<0.05),各康柏西普组能抑制高糖环境下人视网膜微血管内皮细胞VEGF的分泌;Western blot示:25μg/L、2.5μg/L康柏西普组occludin蛋白表达量较高糖组高(P<0.05),但0.25μg/L康柏西普与高糖组无明显差异(P>0.05);TER值、HRP通透性检测示:康柏西普能抑制高糖组TER值的降低,并抑制HRP通透性的增加。结论康柏西普对高糖环境下血视网膜内屏障具有一定的保护作用,其机制可能与VEGF有关。

Objective：To investigate the effects of conbercept on human retinal microvascular endothelial cells （ hRMECs）barrier function and expression of VEGF in high glucose environment. Methods：The hRMECs were divided into 5 groups：control group（ Group N）,high glucose group（ Group H）,and treatment groups（ high glucose with different concentration of conbercept）. The examination of vascular endothelial growth factor（ VEGF）was done by enzyme linked immunosorbent assay（ ELISA）. The protein level of occludin in each group was detected by western blot. Transendothelial electrical resistance（ TER）and the permeability of horseradish peroxidase（ HRP）were detected to evaluate the stability of barrier function of hRMEC. Results：The level of VEGF in group H is significantly different from that in group N（ P﹤0. 05）. However,this effect was significantly inhibited by conbercept treatment（P﹤0. 05）. The concentration of 25μg/L, 2. 5μg/L conbercept increased protein level of occludin compared with group H,which was not found in the concentration of 0. 25μg/L conbercept. Conbercept suppressed not only the reducation of TER value but also the increase in permeability of HRP. Conclusion：Conbercept can protect the barrier function of hRMECs against high glucose,whose mechanism might be associated with VEGF.