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?
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±