Li Longjie, Jiang Liping, Geng Chengyan, Cao Jun, Zhong Laifu
Department of Toxicology, Dalian Medical University, No. 9, West Segment of South lushun Road, Lushunkou District, Dalian 116044, Liaoning, PR China.
Free Radic Res. 2008 Apr;42(4):354-61. doi: 10.1080/10715760802008114.
This study evaluated the role of oxidative stress in acrolein-induced DNA damage, using HepG2 cells. Using the standard single cell gel electrophoresis (SCGE) assay, a significant dose-dependent increment in DNA migration was detected at lower concentrations of acrolein; but at the higher tested concentrations, a reduction in the migration was observed. Post-incubation with proteinase K significantly increased DNA migration in cells exposed to higher concentrations of acrolein. These results indicated that acrolein caused DNA strand breaks and DNA-protein crosslinks (DPC). To elucidate the oxidatively generated DNA damage mechanism, the 2,7-dichlorofluorescein diacetate (DCFH-DA) and o-phthalaldehyde (OPT) were used to monitor the levels of reactive oxygen species (ROS) and glutathione (GSH), respectively. The present study showed that acrolein induced the increased levels of ROS and depletion of GSH in HepG2 cells. Moreover, acrolein significantly caused 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) formation in HepG2 cells. These results demonstrate that the DNA damage induced by acrolein in HepG2 cells is related to the oxidative stress.
本研究使用HepG2细胞评估了氧化应激在丙烯醛诱导的DNA损伤中的作用。采用标准单细胞凝胶电泳（SCGE）试验，在较低浓度的丙烯醛下检测到DNA迁移有显著的剂量依赖性增加；但在较高测试浓度下，观察到迁移减少。用蛋白酶K孵育后，暴露于较高浓度丙烯醛的细胞中的DNA迁移显著增加。这些结果表明，丙烯醛导致DNA链断裂和DNA-蛋白质交联（DPC）。为了阐明氧化产生的DNA损伤机制，分别使用2,7-二氯荧光素二乙酸酯（DCFH-DA）和邻苯二甲醛（OPT）监测活性氧（ROS）和谷胱甘肽（GSH）的水平。本研究表明，丙烯醛诱导HepG2细胞中ROS水平升高和GSH消耗。此外，丙烯醛显著导致HepG2细胞中8-氧代-7,8-二氢-2'-脱氧鸟苷（8-oxodGuo）形成。这些结果表明，丙烯醛在HepG2细胞中诱导的DNA损伤与氧化应激有关。
