【摘要】 目的　探讨在紫外线诱导的人晶状体上皮细胞DNA损伤修复过程中端粒酶活性和氧化损伤蛋白的变化和作用。方法　采用照射剂量为0 0(对照组)及0 .5、1. 5、2 .5、3 .5、5 .0、7 .5、10 0mJ/cm2 (实验1~7组)的紫外线照射人晶状体上皮细胞,采用端粒酶重复序列扩增酶联免疫吸附测定法检测端粒酶活性,采用免疫印迹法检测p53、DNA损伤诱导生长停顿基因(GADD45)、增殖细胞核抗原(PCNA)及细胞周期依赖激酶抑制剂p16的蛋白表达水平。结果　对照组和实验1~7组的端粒酶活性呈上调趋势, 8组比较差异有统计学意义(F=45. 65,P<0 .01 );对照组和实验1 ~7组的p53、GADD45、PCNA、p16的蛋白表达水平均呈上升趋势, 8组比较差异均有统计学意义(F=13 .015 52, 41 241 .40, 25. 084 19, 44 331 .72;P<0. 01)。结论　在紫外线诱导的人晶状体上皮细胞DNA损伤修复过程中,端粒酶活性上调,氧化损伤蛋白的表达水平上升。端粒酶活性的上调既加强了保护细胞的作用,又增强了增殖作用;氧化损伤蛋白表达水平的提高在紫外线诱导的人晶状体上皮细胞DNA损伤修复过程中发挥关键作用;端粒酶活性上调与氧化损伤蛋白表达水平提高有关。更多还原

【Abstract】 Objective To investigate the changes and the role of telomerase activity and other stress-related proteins in the process of UV-induced DNA damage and repair in human lens epithelial cells. Method Human lens epithelial cells were irradiated at UV-doses 0.0(control group) and 0.5, 1.5, 2.5, 3.5, 5.0, 7.5, 10.0 mJ/cm 2(treated 1-7group). Telomerase activity was determined by Telomerase Repeat Amplification Protocol-Enzyme Linked Immunosorbent Assay(TRAP-ELISA),p53,growth arrest and DNA damage inducible(GADD45),proliferating cell nuclear antigen(PCNA) and p16 protein levels were analyzed by Western blotting. Result Telomerase activity in control group and treated 1-7group showed increased tendency, the differences of telomerase activity in 8 groups were significantly ( P <0.01).The expression of p53,GADD45,PCNA,p16 proteins showed increased tendency in experimental group, comparing with the control group, there were significant difference ( P <0.01). Conclusion During UV-induced DNA damage and repair in human lens epithelial cells, telomerase activity was upregulated and the expression of stress-related proteins levels was increased. Upregulated telomerase activity may play both a protective and a proliferative role in human lens epithelial cells. Increased stress-related proteins level is critic in UV-induced DNA damage and repair in human lens epithelial. Increased telomerase activity is associated with increased levels of the stress-related proteins.更多还原