摘要
目的测定αΒ-晶状体蛋白在体外培养人视网膜色素上皮细胞(retinal pigment epithelial cell,RPE)中的表达,探讨αΒ-晶状体蛋白对体外培养人RPE细胞增殖的影响。方法传代培养人A-RPE19细胞系,免疫荧光技术检测αΒ-晶状体蛋白在RPE细胞中的表达。用不同稀释浓度(体积比分别为1/102、1/103、1/104、1/105、1/106)的αΒ-晶状体蛋白以及其抗体干预各组(对照组、蛋白组、抗体组)细胞,MTT方法检测不同时间点各组细胞光密度值,流式细胞术检测各组细胞不同时间点的周期及凋亡情况。结果αΒ-晶状体蛋白在体外培养人RPE细胞质中表达;在48 h,浓度为1/104～1/102时,3组细胞的光密度值差异有统计学意义(P<0.05);蛋白组与对照组相比,0～48 h蛋白组的G1期比例减小,S、G2期比例增加,抗体组与对照组相比,G1期比例增加,S、G2期比例减少(P<0.05,P<0.01);48 h时3组之间凋亡细胞的构成比差异有统计学意义(P<0.05)。结论αΒ-晶状体蛋白在体外培养人RPE细胞质中表达,并对体外培养人RPE细胞增殖起明显促进作用。
Objective To determine the expression of αB-crystallin in human retinal pigment epithelial cells （RPEC）, and to investigate the effect of αB-crystallin on RPEC proliferation in vitro. Methods A-RPE19 cells were subcultured and the expression of cLB-crystallin was detected using immunofluorescence technique. cLB-crystallin in different concentrations （V/V： 1/102, I/I03, 1/104, 1/105 and 1/106） and its antibody were used for treating different groups of cells, and then OD values and cell cycle together with cell apoptosis at different time points were measured by MTY assay and flow cytometry, respectively. Results c（B-crystallin was found to be expressed in the cytoplasm of human RPEC cultured in vitro. There were statistical significance （ P 〈 0.05 ） in cell OD values among the three groups at 48 h when the concentration was 1/104 - 1/102. The proportion of cells at G1 phase decreased while the proportion of those at S and G2 phases increased in αB- crystallin group as compared with the control group （0 -48 h）. The proportion of ceils at G1 phase increased while the proportion of those at S and G2 phases decreased in αB-crystallin antibody group as compared with the control group （0-48 h）. The ratios of apoptotic cells among the three groups were significantly different at 48 h （P 〈 0.05）. Conclusion Our study verified the expression of （αB-crystallin in the cytoplasm of human RPEC in vitro, and （αB-crystallin plays a distinct role in promotinz RPEC oroliferation.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第3期248-250,共3页
Journal of Third Military Medical University
基金
重庆市卫生局资助项目(2011-1-029)~~
