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本研究旨在探讨线粒体DNA氧化损伤热点的形成是否与该点DNA修复速率慢有关 .BRL 3A细胞暴露于 5 0mmol·L- 1过氧化氢 (H2 O2 ) 30min后 ,分别于 0 ,1,4及 2 4h收获细胞 ,应用连接介导聚合酶链反应分析线粒体DNA氧化损伤热点的修复百分率并与线粒体DNA氧化损伤的总修复百分率比较 ,通过狭缝印迹法 ,比较了各时间点细胞内p5 3/线粒体DNA比率来验证其修复的可靠性 .结果表明 :在 4及2 4h ,热点的修复速率分别为 5 .2 %和 4 2 .1% .而线粒体DNA的总修复百分率分别为 76 .9%和 84 .1% ,后者与前者相比 ,其修复速率快 1～ 14倍左右 .应用狭缝印迹发现各时间点细胞内p5 3/线粒体DNA比率无明显变化 .因此 ,线粒体DNA氧化损伤热点的形成与其修复速率较慢有关 .狭缝印迹结果表明所观测的线粒体DNA的修复是可信的
The purpose of this study was to investigate whether the formation of mitochondrial DNA oxidative damage hot spots is related to the slow DNA repair rate at this point.BRL 3A cells exposed to 50 mmol·L -1 H2O2 for 30 min, And the cells were harvested at 24 hours. The percentage of repair of mitochondrial DNA oxidative damage by ligation-mediated polymerase chain reaction (PCR) was compared with that of mitochondrial DNA oxidative damage. By slit-blotting, the intracellular p5 3 / Mitochondrial DNA ratio to verify the reliability of its repair.The results showed that the repair rates of hot spots were 5.2% and 42.1% at 4 and 24 h respectively, while the total repair rates of mitochondrial DNA were 76.9 % And 84.1%, respectively, the latter being about 1 to 14 times faster than the former, and no significant change in the ratio of p5 3 / mitochondrial DNA was observed at each time point using slit blotting. Therefore, mitochondrial DNA oxidative damage The formation of hot spots is associated with a slower rate of repair.The slit blotting results indicate that the observed mitochondrial DNA repair is credible