不同体积分数氧对胎鼠肺泡Ⅱ型上皮细胞体外生长状态的影响

Effects of different state of oxidative stress on the fetal rat alveolar typeⅡ epithelial cells in vitro

目的:探讨不同氧体积分数状态下,胎鼠原代肺泡Ⅱ型上皮细胞(Alveolar typeⅡepithelial cell,AECⅡ)体外生长状态的变化。方法:原代培养胎鼠AECⅡ并用电镜鉴定;建立氧体积分数0.95组(95%O2)、氧体积分数0.4组(40%O2)和空气组(21%O2)3种氧化损伤性细胞模型,各自暴露12、24 h和48 h;用倒置相差显微镜观察各组细胞的形态变化;四甲基偶氮唑盐反应比色法(MTT法)检测细胞存活率,流式细胞术检测细胞凋亡率和细胞内活性氧(Reactive oxygen species,ROS)浓度。结果:与空气组相比,氧体积分数0.4组在12、24 h存活率和凋亡率均无统计学差异,但48 h后出现统计学差异,ROS在24 h即出现统计学差异;氧体积分数0.95组每组均和空气组有统计学差异。此外,氧体积分数0.4组细胞的形态变化较氧体积分数0.95组损伤轻,但仍重于空气组。结论:体外40%O2的氧刺激在超过48 h以后,AECⅡ也会出现相应的氧化损伤。

Objective：To investigate effects of different state of oxidative stress on fetal rats alveolar type Ⅱ epithelial cells（AECⅡ）cultured in vitro.Methods：AECⅡwere primarily cultured;then randomly divided into three groups of oxidative damage with the oxygen concentration 0.95（95% O2）,0.4（40% O2）and room air（21% O2）.Each group was exposed for 12,24 h and 48 h respectively.Then cell morphological change was observed by inverted microscope;and cell survival rate was determined by MTT assay,apoptosis rate and intracellular reactiveosygenspecies（ROS）concentration were determined by flow cytometry.Results：Compared to room air group,survival rate and apoptosis rate of oxygen concentration 0.4 group were not significantly diferent after 12 h and 24 h exposure,but the differences were significant after 48 h exposure,and ROS was significantly different after 24 h exposure;there were significant differences between oxyren concentration 0.95 group and roon air group at three time points.In addition,the morphological change of cells in oxygen concentration 0.4 group was less damage than those in oxygen concentration 0.95 group,but still heavier than those in room air group.Conclusion：After stimulation by 40%O2 over 48 h in vitro,corresponding oxidative damage would also appear in AECⅡ.