碱性成纤维细胞生长因子在糖基化终产物作用下视网膜Müller细胞中的表达及对血管内皮生长因子影响

目的 观察糖基化终产物(AGEs)作用下兔视网膜Müller细胞中碱性成纤维细胞生长因子(bFGF)的表达,以及bFGF对该细胞表达血管内皮生长因子(VEGF)的影响,探讨AGEs对于糖尿病视网膜病变(DR)发生发展的可能作用机制.方法 制备牛血清白蛋白AGEs(AGEs-BSA)及其对照物并将其作用于体外培养的兔视网膜Müller细胞,采用免疫细胞化学(ICC)方法 半定量检测不同时间点(1d、3d、6d、9d)Müller细胞bFGF表达变化.利用外源性bFGF(0.1、1.0、10.0、50.0、100.0)ng/mL干预Müller 细胞,采用ICC方法 半定量检测不同时间点(1d、3d、6d、9d)M üller细胞VEGF的表达变化.结果 AGEs作用下兔视网膜M üller细胞bFGF的表达增高(P<0.05或P<0.01)且具有一定的时间和浓度依赖性.外源性bFGF可上调视网膜M üller细胞VEGF的表达,且具有一定的浓度依赖性及时限性.结论 AGEs可以上调视网膜Mü ller细胞表达bFGF,而bFGF可以上调Müller细胞表达VEGF,推测AGEs可能通过增加bFGF的表达,以及通过分泌的bFGF间接促进VEGF的表达,从而在DR形成过程中起重要作用.

Abstract：

Objective To investigate the effect of advanced glycosylation end products (AGEs) on expression of basic fibroblast growth factor (bFGF) in rabbit retinal Müller cells, also the effect ofbFGF on expression of vascular endothelial growth factor (VEGF) in rabbit retinal Müller cells in vitro. Methods Rabbit retinal Müller cells were cultured first, then AGEs-BSA and its control were prepared. Müller cells were treated with 5 different concentration series of AGEs-BSA and AGEs-BSA control for 1, 3, 6 and 9 days, while blank control group was incubated without any intervention. Then bFGF expression in Müller cells was half-quantitatively identified by immunocytochemistry (ICC). Cultured rabbit Müller cells were also treated with bFGF of 5 concentrations (0.1, 1.0, 10.0, 50.0, 100.0) ng/mL for 1, 3, 6, 9 days, while blank control group was incubated without any intervention. Then VEGF expression in Müller cells was half-quantitatively identified by ICC. Results Comparing with control group, AGEs-BSA evoked a time and concentration-dependent increase ofbFGF expression on cultured retinal Müller cells (P ＜0. 05 or P ＜0. 01). And comparing with blank control group, bFGF also evoked a time and concentration-dependent increase of VEGF expression on retinal Mi ller cells in a certain range. Conclusions AGEs up-regulates the expression ofbFGF, which can up-regulate the expression of VEGF in Müller cells. These results indicate that AGEs might promote the progress of DR.