中国当代儿科杂志Issue(3)：275-280,6.DOI:10.7499/j.issn.1008-8830.2015.03.016
PKCβ/P66Shc氧化应激通路在高氧诱导人肺泡上皮细胞活性氧簇产生中的作用
Roles of PKCβ/P66Shc oxidative stress signal pathway in mediating hyperoxia-induced ROS production in alveolar epithelial cells
摘要
Abstract
ObjectiveTo explore the roles of PKCβ/P66Shc oxidative stress signal pathway in mediating hyperoxia-induced reactive oxgen species (ROS) production in alveolar epithelial cells (A549) and the protective effects of PKCβ inhibitor on hyperoxia-induced injuries of alveolar epithelial cells.MethodsA549 cells were culturedin vitro and randomly divided into three groups: control, hyperoxia and PKCβ inhibitor LY333531 treatment. The hyperoxia group was exposed to a mixture of O2 (950 mL/L) and CO2 (50 mL/L) for 10 minutes and then cultured in a closed environment. The LY333531 group was treated with PKCβ inhibitor LY333531 of 10 µmol/L for 24 hours before hyperoxia induction. Cells were collected 24 hours after culture and the levels of PKCβ, Pin1, P66Shc and P66Shc-Ser36 were detected by Western blot. The intracellular translocation of P66Shc, the production of ROS and cellular mitochondria membrane potential were measured using the confocal microscopy.ResultsCompared with the control group, the levels of PKCβ, Pin1, P66Shc and P-P66Shc-Ser36 in A549 cells 24 hours after culture increased signiifcantly in the hyperoxia group. These changes in the hyperoxia group were accompanied with an increased translocation rate of P66Shc from cytoplasm into mitochondria, an increased production of mitochondrial ROS, and a reduced mitochondrial membrane potential. Compared with the hyperoxia group, the levels of Pin1, P66Shc and P66Shc-Ser36 in A549 cells,the translocation rate of P66Shc from cytoplasm into mitochondria and the production of mitochondrial ROS decreased significantly, while the mitochondrial membrane potential increased significantly in the LY333531 treatment group. However, there were signiifcant differences in the above mentioned measurements between the LY333531 treatment and control groups.ConclusionsHyperoxia can increase the expression of PKCβ in alveolar epithelial cells and production of mitochondrial ROS and decrease mitochondrial membrane potential. PKCβ inhibitor LY333531 can partially disrupt these changes and thus alleviate the hyperoxia-induced alveolar epithelial cell injury.关键词
PKCβ/P66shc/活性氧簇/氧化应激/肺泡上皮细胞Key words
PKCβ/P66Shc/Reactive oxgen species/Oxidative stress/Alveolar epithelial cell引用本文复制引用
车忠丽,董文斌,李清平,雷小平,康兰,郭琳,翟雪松,王胜会,陈枫..PKCβ/P66Shc氧化应激通路在高氧诱导人肺泡上皮细胞活性氧簇产生中的作用[J].中国当代儿科杂志,2015,(3):275-280,6.基金项目
中华儿科杂志第二届双鹤珂立苏科研基金；四川省教育厅科研基金（08ZA150）；四川省卫生厅科研基金（90191）。 （）
