摘要目的 探讨高级糖基化终末产物(AGEs)对人视网膜色素上皮(RPE)细胞的作用机制.方法 原代培养人RPE细胞,在传代细胞生长状态良好的情况下,无血清的Dulbecco改良Eagle培养基同步化24 h,进行分组:(1)C组:BSA浓度为0.1 g/L,各作用24、48 h,分别为C1、C2组；(2)NC组:葡萄糖浓度为5.6 mmol/L,各作用24、48 h,分别为NC1、NC2组；(3)A组:AGEs浓度分别为0.1、0.2、0.4 g/L,各作用24、48 h,作用24 h依次为A1组、A2组、A3组,作用48 h依次为A4组、A5组、A6组.免疫组织化学检测AGEs受体(RAGE)、过氧化物酶体增生物激活受体γ辅助激活因子-1α(PGC-1α)和血管内皮生长因子(VEGF)蛋白的表达；激光共聚焦显微镜检测核因子-κB (NF-κB)的活化.通过图像分析软件IPP6.0和统计软件SPSS 17.0进行定量分析.结果 A组RPE细胞较C组和NC组RAGE、PGC-1α和VEGF蛋白的表达明显增加,且随着AGEs浓度的升高和刺激时间的延长而增加(F=294.5、228.3、241.5,P＜0.05)；随着AGEs浓度的升高和刺激时间的延长,NF-κB的活化增加.结论 AGEs的堆积可导致其受体RAGE在RPE细胞表达增加,并促进核转录共同激活因子PGC-1α蛋白的表达和NF-cB的活化,促进RPE细胞VEGF的分泌.

abstractsObjective To study the effect of advanced glycosylation end products (AGEs) on human retinal pigment epithelium (RPE) cells.Methods Human primary RPE cells were cultured in basal and different concentrations of AGEs with different times.The cells were divided into several groups as follows:group C (control):bovine serum albumin 0.1 g/L,24 hours (C1) and 48 hours (C2); group NC (normal control):basal culture medium with 5.6 mmol/L of glucose,24 hours (NC1) and 48 hours (NC2) ; group A (AGEs):0.1 g/L,24 hours and 48 hours,A1 and A4 ; 0.2 g/L,24 hours and 48 hours,A2 and A5 ; 0.4 g/L,24 hours and 48 hours,A3 and A6.Immunohistochemistry analysis was used to study the protein expression of receptor for AGEs (RAGE),peroxisome proliferative-activated receptor-gamma coactivator-1 alpha (PCG -1α) and vascular endothelial growth factor (VEGF) protein.The activation of nuclear factor-kappa B (NF-κB) was detected by confocal microscope.Software IPP6.0 and SPSS 17.0 were used to analyze the quantitation data.Results Immunohistochemistry analysis showed that RAGE protein,PGC-lα protein and VEGF protein were basally secreted in RPE cells,but AGEs can obviously increases the expression level of these proteins (F =294.5,228.3,241.5; P ＜ 0.05).Confocal microscope demonstrated that AGEs increased the activation of NF-κB significantly.Conclusion Accumulation of AGEs can stimulate the expression of RAGE protein,PGC-1α protein and VEGF protein,activation of NF-κB and induce apoptosis of RPE cells.