CAJ | 학술논문

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（WesternBlot）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。
목적：탐토강백서보대고당배경하인시망막미혈관내피세포병장공능급VEGF표체적영향。방법장인시망막혈관내피세포분위정상조（5.5 mol/L포도당）、고당조（25 mol/L포도당）、고당+불동농도（25μg/L、2.5μg/L、0.25μg/L）적강백서보조。응용매련면역흡부측정（ ELISA）검측각조세포상청액중혈관내피생장인자（ VEGF）적농도；과막전조（ TER）급랄근과양화물매（ HRP）통투성적검측，이평개세포병장적은정성；단백질인적법（WesternBlot）검측긴밀련접상관단백occludin적표체。결과 ELISA결과시：고당조분비적VEGF비정상조고，증가차이구유통계학의의（P﹤0.05），각강백서보조능억제고당배경하인시망막미혈관내피세포VEGF적분비；Western blot시：25μg/L、2.5μg/L강백서보조occludin단백표체량교고당조고（ P﹤0.05），단0.25μg/L강백서보여고당조무명현차이（ P﹥0.05）；TER치、HRP통투성검측시：강백서보능억제고당조TER치적강저，병억제HRP통투성적증가。결론강백서보대고당배경하혈시망막내병장구유일정적보호작용，기궤제가능여VEGF유관。
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.