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目的:研究Pten缺失后小鼠胚胎成纤维细胞抗氧化能力、细胞内活性氧(reactive oxygen species,ROS)水平、脂质以及DNA氧化损伤水平的变化。方法:采用超氧化物歧化酶(superoxide dismutase,SOD)活力检测、ROS荧光发光分析、丙二醛(malondialdehyde,MDA)含量检测和.γ-H2AX免疫荧光染色技术,分别比较Pten~(+/+)MEFs与Pten~(-/-)MEFs细胞SOD活力、ROS、MDA和γ-H2AX水平的差异。结果:Pten~(-/-)MEFs细胞中SOD活力明显减弱,ROS、MDA和γ-H2AX水平均显著增高。结论:Pten可能通过调控细胞抗氧化能力影响ROS水平,从而拮抗脂质和DNA氧化损伤以及由此产生的染色体不稳定性。
OBJECTIVE: To study the anti-oxidative ability, reactive oxygen species (ROS) levels, lipid and DNA oxidative damage of mouse embryonic fibroblasts after Pten deletion. Methods: The activity of superoxide dismutase (SOD), the fluorescence of ROS, the content of malondialdehyde (MDA) and the activity of γ-H2AX immunofluorescence staining were compared. The levels of Pten ~ (+ / + ) MEFs and Pten ~ (- / -) MEFs cells, SOD, MDA and γ-H2AX levels. Results: The activity of SOD, the content of ROS, MDA and γ-H2AX in Pten ~ (- / -) MEFs cells were significantly decreased. CONCLUSION: Pten may affect ROS levels by regulating cellular antioxidant capacity, thereby antagonizing lipid and DNA oxidative damage and resulting chromosomal instability.