αΒ-晶状体蛋白促进体外培养人RPE 细胞的增殖.docVIP

αΒ-晶状体蛋白体外培养人RPE细胞增殖 400016重庆，重庆医科大学附属第一医院眼科） 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，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RPE-19细胞免疫荧光技术检测αΒ-晶状体蛋白在RPE细胞中的表达。用不同稀释浓度(1/102、1/103、1/104、1/105、1/106)的αΒ-晶状体蛋白以及抗体相应细胞，MTT方法检测组细胞，流式细胞技术检测各组细胞的周期及凋亡情况。结果αΒ-晶状体蛋白体外培养人RPE细胞中在h，浓度为1/104～1/102时，3组细胞的光度值差异有统计学意义（＜0.05）蛋白组与对照组相比48h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期比例减少凋亡细胞的构成比差异有统计学意义（＜0.05）结论证实αΒ-晶状体蛋白在体外培养人RPE细胞表达，并对体外培养人RPE细胞增殖起明显促进作用。 关键词αΒ-晶状体蛋白；人视网膜色素上皮细胞；增殖αΒ-crystallin played a role in promoting RPE cells proliferation Tang Derong, Zhang Xuedong,Cheng Xiaoli,Shen Qiang,Li Yingyuan (Department of Ophthalmology，First Affiliated Hospital,Chongqing Medical University,Chongqing,400016,China) Abstract] Objective To determine the expression of αΒ-crystallin in human retinal pigment epithelial cells（RPE）and to investigate the effects of αΒ-crystallin on RPE proliferation cultured in vitro.Methods ARPE-19 cells were subcultured and the expression of αΒ-crystallin was detected using immunofluorescence technique. αΒ-crystallin in different concentrations (1/102,1/103,1/104,1/105,1/106:V/V) and its antibody were used for treating different groups of cells , then OD values and cell cycle together with cell apoptosis at different times were measured by MTT assay and flow cytometry, respectively. Results αΒ-crystallin were found to be expressed in the cytoplasm of human RPE cells cultured in vitro；There were statistically significance （P＜0.05）in cell OD values among the three groups at 48h when the concentration used was 1/104-1/102；The proportion of G1 phase decreased while the proportion of S,G2 phase increased in αΒ-crystallin group compared to the control group (0-48h);The proportion of G1 phase increased while the proportion of S,G2 phase decreased in αΒ-crystallin antibody group compared to the control group (0-48h)；The constituent ratios of apoptotic cells among the three groups were statistically significant at 48h（P＜0.05）.Conclusion