摘要
目的探讨高氧对胎鼠肺泡Ⅱ型上皮细胞（type11alveolarepithelialcells，AECⅡ）的活力、生长情况、增殖和凋亡的影响。方法取孕19d的胎鼠肺组织，利用差速贴壁法进行AECⅡ的原代培养及纯化，将AECⅡ随机分为高氧组及空气组。空气组细胞置于5％CO2孵育箱中培养，高氧组细胞置于5％CO2＋95％O2的混合气体孵育箱中培养。在培养的2、4、6、8d分别观察细胞的生长情况，检测细胞活力、细胞周期、细胞凋亡情况。时间点与培养组间的交互作用采用析因设计的方差分析。2组资料比较采用独立样本t检验，多组资料比较采用单因素方差分析，组间两两比较采用Bonferroni方法。结果（1）细胞生长状况：高氧组培养2、4、6和8d时细胞数随培养时间延长逐渐减少，分别为（7．29±0．43）×10^5/ml、（2．68±0．37）×10^5／ml、（0．23±0．10）×10^5／ml和（0．00±0．00）×10^5／ml，低于相应时间点空气组[分别为（10．41±0．24）×10^5／ml、（27．90±1．91）×10^5／ml、（27．12±0．85）×10^5／ml和（26．29±1．59）×10^5／ml]，差异均有统计学意义（t分别为10．992、38．912、94．166和49．696，P均=0．000）。（2）细胞活力：高氧组在2、4、6d时细胞活力逐渐下降，活细胞率分别为（79．00±0．71）％、（52．80±1．14）％和（31．60±1．52）％，均低于相应时间点空气组，分别为（97．00±0．71）％、（97．20±0．84）％和（95．00±0．71）％，差异均有统计学意义（t分别为31．213、70．519和8乱722，P均-0．000）。（3）细胞周期变化：高氧组G1期和s期细胞百分率在4和6d时与2d时比较均有升高，且高于空气组，差异均有统计学意义（P〈0．05）。（4）细胞凋亡变化：高氧组Annexin—V＋／PI亚群在4d时所占比例最高，为（23．89±0．52）％，高于2d和6d时[（21．32±0．43）％和（1．47±0．61）％]；Annexin—V＋／PI＋亚群细胞所占比例6d时最高，为（53．92±1．64）％，其次是4d时[（45．03±1．01）％]，最低是2d时[（12．17±0．60）％]；高氧组各时间点AnnexinV＋／PI和AnnexinV。／PI＋亚群所占比例与对照组比较，差异均有统计学意义（P〈0．05）。结论高氧对AECⅡ的生长、活力、增殖能力、细胞周期具有明显的抑制作用，对细胞凋亡有促进作用。
Objective To investigate the effect of hyperoxia on growth of type Ⅱ alveolar epithelial cells （AECⅡ）. Methods Lungs of fetal rats at 19 days of pregnancy were collected, and AEC Ⅱ was isolated and cultured by differential adherence method. Cells were randomly divided into air group and hyperoxia group. In air group, cells were cultured in 5% CO2 incubator. And cells in hyperoxia group were cultured in 5% CO2＋95%O2 incubator. The growth, activity, cell cycle, cell apoptosis of AEC Ⅱ were observed at 2, 4, 6 and 8 days of culture. The interaction between different time and groups were analyzed by ANOVA of factorial design. Comparison of means was done by two-sample independent t test and one-way analysis of variance. Bonferroni correction was used during the comparisons. Results （1） Cell growth situation： in hyperoxia group, cell number was decreased from2 hto 8 h [（7.29±0.43） X 10^5/ml, （2.68±0.37））〈 105/ml, （0.23±0. 10） × 10^5/ml and （0. 00±4-0. 00） ×10^5/ml], and lower than those in air group [（10.41±0.24））〈105/ml, （27.904- 1.91） X lOS/ml, （27.12±0.85））〈 105/ml and （26.29-1.59）× 10^5/ml]（t= 10. 992, 38. 912, 94. 166 and 49. 696,P=0. 000 respectively）o （2） Cell activity： the living cells ratio in hyperoxia group at 2 d （79.00±0.71） %], 4 d [（52.80±1.14）%/00] and 6 d [（31. 60± 1.52）%] was lower than those E（97.00±0.71）%, （97.20±0.84）% and （95.00±0.71）%] in air group （t=31.213, 70.519 and 84. 722, P=0. 000 respectively）. （3） Cell cycle., the cell ratios of G1 phase and S phase in hyperoxia group at day 4 [（66.82±1.20） % and （27.31±1.16）%] and day 6 [（70. 22±1.27） %and （30. 31± 1.40） %] were significantly higher than that at day 2 and that in air group （P〈0.05 respectively）. （4） Cell apoptosis： in hyperoxia group, the cell ratio of Annexin-V＋/PI subgroup at 4 h was the highest [（23.89±0.52）%],followed by those at day 2 and 6 [（21.32±0.43）% and （1.47± 0.61）]. While the cell ratio of Annexin-V＋/PI＋ was the highest at 6 h [（53.92±1.64）], followed by those at 4 h and 2 h [（45. 03±1.01）] and （12.17±0.60） , which were all different with those in air group（P〈0.05 respectively）. Conclusions Hyperoxia might inhibit cell activity and cell cycle of AEC Ⅱ and promote apoptosis.
出处
《中华围产医学杂志》
CAS
北大核心
2012年第1期30-36,共7页
Chinese Journal of Perinatal Medicine
基金
国家自然科学基金项目（30772036）
山东省科技攻关项目（2009GG20002055）
山东省优秀中青年科学家科研奖励基金（BS2010YY003）
关键词
高氧症
肺泡
上皮细胞
细胞生长过程
细胞凋亡
Hyperoxia
Pulmonary alveoli
Epithelial cells
Cell growth processes
Apoptosis
