论文部分内容阅读
目的:研究糖基化终产物(advanced glycationendproducts, AGEs)在年龄相关性黄斑变性发生中的作用。方法:使用lycopersiconesculentum agglutinin包被磁珠的改良方法分离牛脉络膜微血管内皮细胞(bovine choroidalendothelialcells, CEC),采用抗VIII因子免疫细胞化学法和摄取低密度脂蛋白(dil-acetylated low-density lipoprotein,dil-ac-LDL)法进行细胞鉴定。将50g/L牛血清白蛋白和150g/L葡萄糖在37℃孵育6wk 以制备AGEs,通过抗-AGEs 抗体点杂交法定性分析。用MTT 法(3,(4,5-dimethylthiazol-2-yl)-2,5- diphenyl te-trazolium bromide assay)测定CEC 增生,用Vit-rogen凝胶系统观察管腔形成情况。结果:培养细胞中90%以上呈因子阳性,并具有摄取dil-ac-LDL的能力,表明具有内皮细胞的特征。制备的AGEs 与抗-AGEs 抗体具有亲和力。浓度在62.5～500mg/L范围内的AGEs 处理CEC 3d 后,细胞增生呈剂量依赖性增加。碱性成纤维细胞生长因子(basic fibroblastgrowth factor, bFGF)可明显增强CEC 中管腔样结构的形成(P <0.05),但500mg/L和50mg/L的AGEs 对CEC 管腔形成无明显影响(P >0.05)。结论:AGEs 可以促进CEC 的增生,但对其管腔形成无影响。提示AGEs 通过增强CEC 增生,可能是渗出型AMD 发病中潜在的启动因子之一?
Objective: To investigate the role of advanced glycation end products (AGEs) in the development of age-related macular degeneration. METHODS: Bovine choroid aldothelial cells (CECs) were isolated by modified lycopersiconesculentum agglutinin-coated magnetic beads. Anti-factor VIII immunocytochemistry and dil- acetylated low-density lipoprotein (dil- ac-LDL) method for cell identification. AGEs were prepared by incubating 50 g / L bovine serum albumin and 150 g / L glucose at 37 ° C for 6 weeks, and qualitatively analyzed by dot-blot hybridization with anti-AGS antibodies. CEC hyperplasia was determined by MTT method (3, 4,5-dimethylthiazol-2-yl) -2,5-diphenyl te-trazolium bromide assay, and lumen formation was observed by Vitro-rogen gel system. RESULTS: More than 90% of the cells in the culture were positive for the factor and had the capacity to ingest dil-ac-LDL, indicating endothelial cell characterization. The prepared AGEs have affinity with anti-AGS antibodies. AGEs in the concentration range of 62.5 ~ 500mg / L CEC 3d after treatment, cell proliferation in a dose-dependent manner. Basic fibroblast growth factor (bFGF) significantly enhanced the formation of luminal structures in CECs (P <0.05), but no significant effect of 500 mg / L and 50 mg / L of AGEs on luminal CEC formation was observed P> 0.05). Conclusion: AGEs can promote the proliferation of CEC, but have no effect on its lumen formation. Tip AGEs by enhancing CEC proliferation may be one of the potential promoter in the pathogenesis of exudative AMD?