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首页|过氧化氢诱导的人正常和早老细胞DNA损伤和修复研究

过氧化氢诱导的人正常和早老细胞DNA损伤和修复研究

the study of hydrogen peroxide induced DNA damage and recovery in normal aging and premature aging human cells

中文摘要英文摘要

为研究人类衰老机制,采用免疫荧光技术和彗星电泳技术,分别检测3组不同群体倍增数(population doubling, PD)的人正常二倍体成纤维细胞BJ(青年组第14代,成年组第30代,衰老组第45代)和2组不同群体倍增数的人HGPS(Hutchinson-Gilford progeria syndrome)早老细胞(青年组第8代,衰老组第17代)的DNA基础损伤程度,然后研究过氧化氢导致的以上细胞组DNA损伤和去除致损伤因素正常培养后的修复水平,最后检测这些细胞组在沉默DNA损伤修复蛋白XPA(Xeroderma pigmentosum group A)表达前后的修复能力。研究显示,BJ细胞衰老组DNA损伤程度较高,与成年组相比,对DNA损伤诱导因子更加敏感。同样,成年组与青年组相比,对损伤诱导因子也更敏感。HGPS细胞青年组的DNA基础损伤程度即已达到衰老BJ细胞类似或更高水平,并且与BJ细胞具有相同的一致的DNA损伤年龄变化趋势。本实验证实XPA功能异常抑制了HGPS细胞的损伤修复,经siRNA沉默其表达后可部分恢复其修复能力,对正常衰老BJ细胞则没有影响。随年龄增长无论正常还是早老细胞,DNA损伤程度增加,修复效率降低。

In the present study, the immunofluorescent assay and Comet assay were used to estimate basal DNA damage in normal aging BJ cells and premature aging HGPS cells, which were divided into three and two distinct population doubling( PD) number groups (BJ 14, 30 , 45and HGPS 8 , 17) respectively. The DNA damage induced by hydrogen peroxide of these cell populations, as well as their repair activity, was also studied. Finally, the recovery capability before and after the XPA knocked down in these groups was measured. Our results indicate that the normal BJ cells of older PD number showed higher basal levels of DNA damage and were more sensitive to the effects of the DNA-damaging agents than the adult one, who, in turn, was more sensitive than the younger ones. The basal damage level of HGPS cells was same or higher than the older BJ cells. HGPS cells also had the same consistent damage change as BJ cells after treatment. The decline of the repair efficiency with age to the DNA damage induced by the agent was also observed.

所起凤、杜文婷、杨鸣鸣、范雪娇、刘戟

基础医学细胞生物学分子生物学

NA损伤修复彗星电泳免疫荧光过氧化氢

NA damage and repairComet assayimmunofluorescent assayhydrogen peroxide

所起凤,杜文婷,杨鸣鸣,范雪娇,刘戟.过氧化氢诱导的人正常和早老细胞DNA损伤和修复研究[EB/OL].(2010-12-22)[2025-08-24].http://www.paper.edu.cn/releasepaper/content/201012-849.点此复制

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