高氧对人肺泡上皮细胞凋亡的影响及其作用机制  

Effects of hyperoxia on apoptosis of human alveolar epithelial cells and the mechanism

作　　者：王霞 董文斌 李清平 康兰 雷小平 翟雪松 赵帅 

机构地区：[1]四川医科大学附属第一医院,四川泸州646000

出　　处：《山东医药》2016年第3期14-16,共3页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81571480);中华儿科杂志第二届双鹤珂立苏科研基金资助项目(cjp2011-009);四川省教育厅科研基金资助项目(08ZA150);四川省卫生厅科研基金资助项目(90191)

摘　　要：目的观察高氧对人肺泡上皮细胞(HPAEC)凋亡的影响,并探讨其机制。方法取对数生长期HPAEC细胞,随机分为高氧组和对照组。对照组细胞不作处理,放置于50 m L/L CO2培养箱中培养;高氧组细胞换液1次后,以3 L/min速度通入含900 m L/L O2和50 m L/L CO2高纯混合气,通入时间10 min;两组细胞均培养24 h。采用流式细胞仪检测细胞凋亡率,倒置相差显微镜下观察细胞形态学变化,免疫共聚焦检测细胞组蛋白去乙酰化酶(SIRT1)转位情况,Western blotting法检测细胞SENP1蛋白、乙酰化p53蛋白。结果高氧组和对照组细胞凋亡率分别为24.77%±2.17%、5.33%±2.60%,两组比较,P<0.05。高氧组细胞从正常的长梭形、多角形变成圆形、椭圆形,细胞间的间隙增宽,悬浮的细胞较多;对照组细胞大多为长梭形、多角形,贴壁较好,细胞间的间隙较小,悬浮的细胞较少。高氧组和对照组细胞SIRT1蛋白转位率分别为88.89%、16.23%,两组比较,P<0.05。高氧组和对照组细胞SENP1蛋白相对表达量分别为0.76±0.12、0.67±0.02,乙酰化p53蛋白相对表达量分别为0.81±0.07、0.52±0.03,两组比较,P均<0.05。结论高氧可促进HPAEC细胞凋亡,其机制可能是高氧诱导HPAEC细胞SENP1蛋白表达增加,进而SIRT1蛋白从胞核转位到胞质,使乙酰化p53蛋白表达增加。Objective To observe the influence of hyperoxia on the apoptosis of human alveolar epithelial cells （HPAEC）, and to explore its mechanism. Methods The HPAEC cells in the logarithmic phase were randomly divided into the hyperoxia group and control group. The cells of the control group were not processed and were placed in 50 mL/L CO2 incubator. After one time of liquid change, the cells of the hyperoxia group were exposed to a mixture of 02 （ 900 mL/L） and CO2（50mL/L） for 10 minutes with speed of 3 L/rain. After 24 hours, the apoptosis rates of the two groups were detected by flow cytometry, the morphological changes were observed by the inverted phase contrast microscope, the transposition of histone deaceylase （ SIRT1 ） was detected by immunofluorescence and the expression of SENP1 and acety- lated p53 proteins were measured by Western blotting. Results The apoptosis rates of the hyperoxia group and control group were 24.77% ±2.17% and 5.33% ±2.60% , respectively （P 〈0.05）. In the hyperoxia group, the shapes of cells changed from normal long spindle, polygon into the circle or ellipse, the distance between cells was enlarged, and the sus- pension cells were more. In the control group, HPAEC cells were in good condition, closely to each other and the suspen- sion cells were less. The transposition rates of SIRT1 protein in the hyperoxia group and control group were 88.89% and 16.23 %, respectively （ P 〈 0. 05 ）. The expression levels of SENP1 protein in the two groups were separately 0. 76 ± 0. 12 and 0. 67 ±0.02 （P 〈0.05）. The acetylated p53 protein expression of the two groups was 0. 81 ±0.07 and 0. 52±0.03, respectively （P 〈 0.05）. Conclusion Hyperoxia can promote HPAEC cell apoptosis, and its mechanism may be that the expression of SENP1 protein is increased, which promotes the transposition of SIRT1 protein from the nucleus to the cyto-plasm, then the expression of acetylated p53 protein is increased.

关 键 词：高浓度氧 人肺泡上皮细胞 组蛋白去乙酰化酶 SENP1蛋白 乙酰化p53蛋白 

分 类 号：Q255[生物学—细胞生物学]