玻璃膜疣主要成分胆固醇对人视网膜色素上皮细胞金属硫蛋白表达的影响

Effect of cholesterol exposure on the expression of metallothionein in human retinal pigment epithelial cells

目的:研究玻璃膜疣主要成分胆固醇对人视网膜色素上皮细胞ARPE-19中金属硫蛋白表达的影响。方法:体外培养ARPE-19细胞,将细胞分为对照组和胆固醇处理组(2.5 mg/mL),取样时间为0,6,12,24,48,72 h。通过实时定量PCR检测hMT1a,hMT2a和hMT3在转录水平的表达,用蛋白质印迹法检测总金属硫蛋白的表达。结果:在转录水平上hMT1a,hMT2a和hMT3受到胆固醇影响mRNA表达上调,且hMT3上调倍数最大;总金属硫蛋白的蛋白表达随着胆固醇处理时间延长明显增多。结论:玻璃膜疣主要成分胆固醇可以上调人视网膜色素上皮细胞中金属硫蛋白的表达,提示金属硫蛋白表达可受到玻璃膜疣形成起始阶段的刺激,其检测是否能用于年龄相关性黄斑变性的早期发现及早期诊断还需深入探讨。

Objective:To study the effects of cholesterol,the main component of drusen,on the expression of metallothionein of ARPE-19 cells.Methods:The ARPE-19 cell line was cultured in vitro,and the cells were divided into a control group and a cholesterol treatment group(2.5 mg/mL).The treatment time was 0,6,12,24,48,72 hours.Realtime quantitative PCR was used to detect the expression of hMT1a,hMT2a and hMT3 at the mRNA level,and Western blot was used to detect the expression at the protein level.Results:The mRNA expression of hMT1a,hMT2a and hMT3 were up-regulated by cholesterol and the protein expression of total MTs was increased with cholesterol treatment.Conclusion:Cholesterol,the main component of drusen,can up-regulate the expression of metallothionein in human retinal pigment epithelial cells,suggesting that the expression of metallothionine can be stimulated by the initial stage of drusen formation.However,whether its detection can be used for the early detection and early diagnosis of age-related macular degeneration or not still needs to be further explored.