体外培养的人皮肤成纤维细胞老化过程中DNA氧化损伤、8-氧-2′-脱氧鸟苷的积累及修复系统的变化

Takao Kaneko , Shoichi Tahara , Takahiko Taguchi , Hiroshi Kondo
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引用次数: 54

摘要

在体外培养的人皮肤成纤维细胞中,研究了细胞老化对DNA碱基氧化产物8-氧-2′-脱氧鸟苷含量的影响。从胎儿供体皮肤组织培养的TIG-3S细胞DNA中8-氧-2′-脱氧鸟苷含量在增殖停止前立即增加。TIG-114和TIG-104细胞分别来自成人和老年供体的皮肤组织,在体外细胞衰老过程中,8-氧-2′-脱氧鸟苷含量发生了相似的变化。8-oxo-2′-脱氧鸟苷在传代后期细胞中的积累与细胞分裂次数有关,而与培养时间无关。在8-氧-2′-脱氧鸟苷含量增加之前,超氧化物歧化酶和谷胱甘肽过氧化物酶活性增加,而过氧化氢酶活性在传代后期的体外细胞衰老过程中逐渐下降。8-氧-2′-脱氧鸟苷内切酶和DNA聚合酶的活性随着细胞增殖的进展而降低。这些结果表明,在细胞分裂停止前,晚期传代细胞中抗氧化应激的防御系统仍保持足够的活性,但在晚期传代时,抗氧化损伤的修复系统则衰退。超过抗氧化能力和/或修复活性的氧化应激似乎导致8-oxo-2 ' -脱氧鸟苷在传代后期细胞中积累。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Accumulation of oxidative DNA damage, 8-oxo-2′-deoxyguanosine, and change of repair systems during in vitro cellular aging of cultured human skin fibroblasts

Effects of in vitro cellular aging on the content of 8-oxo-2′-deoxyguanosine, a typical oxidation product of DNA bases, were examined in cultured human skin fibroblasts. The 8-oxo-2′-deoxyguanosine content in the DNA of TIG-3S cells established from skin tissues of a fetal donor increased immediately before the cessation of proliferation. TIG-114 and TIG-104 cells established from skin tissues of adult and aged donors, respectively, showed similar changes in 8-oxo-2′-deoxyguanosine content during in vitro cellular aging. The accumulation of 8-oxo-2′-deoxyguanosine in late-passage cells was dependent on the number of cell divisions, and not on the cultivation time. Increases in the activities of superoxide dismutase and glutathione peroxidase were observed prior to the increase in 8-oxo-2′-deoxyguanosine content, while the catalase activity decreased gradually during in vitro cellular aging at late-passage. Furthermore, the activities of 8-oxo-2′-deoxyguanosine endonuclease and DNA polymerases decreased with the progression of proliferation. These results indicate that defense systems against oxidative stress in late-passage cells remain sufficiently active before the cessation of cell division, but that repair systems against oxidative damage decay at late-passage. Oxidative stress beyond the antioxidant capacity and/or repair activity seems to result in an accumulation of 8-oxo-2′-deoxyguanosine in late-passage cells.

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