中华围产医学杂志
中華圍產醫學雜誌
중화위산의학잡지
CHINESE JOURNAL OF PERINATAL MEDICINE
2012年
1期
30-36
,共7页
刘秀香%张海鸿%王克煊%田春梅%封志纯
劉秀香%張海鴻%王剋煊%田春梅%封誌純
류수향%장해홍%왕극훤%전춘매%봉지순
高氧症%肺泡%上皮细胞%细胞生长过程%细胞凋亡
高氧癥%肺泡%上皮細胞%細胞生長過程%細胞凋亡
고양증%폐포%상피세포%세포생장과정%세포조망
Hyperoxia%Pulmonary alveoli%Epithelial cells%Cell growth processes%Apoptosis
目的 探讨高氧对胎鼠肺泡Ⅱ型上皮细胞(typeⅡalveolar epithelial cells,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)×105/ml、(2.68±0.37)×105/ml、(0.23±0.10)×105/ml和(0.00±0.00)×105/ml,低于相应时间点空气组[分别为(10.41±0.24)×l05/ml、(27.90±1.91)×105/ml、(27.12±0.85)×105/ml和(26.29±1.59)×105/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84.722,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Annexin-V+/PI-和Annexin-V+/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.
目的 探討高氧對胎鼠肺泡Ⅱ型上皮細胞(typeⅡalveolar epithelial cells,AECⅡ)的活力、生長情況、增殖和凋亡的影響. 方法 取孕19d的胎鼠肺組織,利用差速貼壁法進行AECⅡ的原代培養及純化,將AECⅡ隨機分為高氧組及空氣組.空氣組細胞置于5％CO2孵育箱中培養,高氧組細胞置于5％CO2 ±95％O2的混閤氣體孵育箱中培養.在培養的2、4、6、8d分彆觀察細胞的生長情況,檢測細胞活力、細胞週期、細胞凋亡情況.時間點與培養組間的交互作用採用析因設計的方差分析.2組資料比較採用獨立樣本t檢驗,多組資料比較採用單因素方差分析,組間兩兩比較採用Bonferroni方法. 結果 (1)細胞生長狀況:高氧組培養2、4、6和8d時細胞數隨培養時間延長逐漸減少,分彆為(7.29±0.43)×105/ml、(2.68±0.37)×105/ml、(0.23±0.10)×105/ml和(0.00±0.00)×105/ml,低于相應時間點空氣組[分彆為(10.41±0.24)×l05/ml、(27.90±1.91)×105/ml、(27.12±0.85)×105/ml和(26.29±1.59)×105/ml],差異均有統計學意義(t分彆為10.992、38.912、94.166和49.696,P均=0.000).(2)細胞活力:高氧組在2、4、6d時細胞活力逐漸下降,活細胞率分彆為(79.00±0.71)％、(52.80±1.14)％和(31.60±1.52)％,均低于相應時間點空氣組,分彆為(97.00±0.71)％、(97.20±0.84)％和(95.00±0.71)％,差異均有統計學意義(t分彆為31.213、70.519和84.722,P均=0.000).(3)細胞週期變化:高氧組G1期和S期細胞百分率在4和6d時與2d時比較均有升高,且高于空氣組,差異均有統計學意義(P＜0.05).(4)細胞凋亡變化:高氧組Annexin-V+/PI亞群在4d時所佔比例最高,為(23.89±0.52)％,高于2d和6d時[(21.32±0.43)％和(1.47±0.61)％]；Annexin-V+/PI+亞群細胞所佔比例6d時最高,為(53.92±1.64)％,其次是4d時[(45.03±1.01)％],最低是2d時[(12.17±0.60)％]；高氧組各時間點Annexin-V+/PI-和Annexin-V+/PI+亞群所佔比例與對照組比較,差異均有統計學意義(P＜0.05). 結論 高氧對AECⅡ的生長、活力、增殖能力、細胞週期具有明顯的抑製作用,對細胞凋亡有促進作用.
목적 탐토고양대태서폐포Ⅱ형상피세포(typeⅡalveolar epithelial cells,AECⅡ)적활력、생장정황、증식화조망적영향. 방법 취잉19d적태서폐조직,이용차속첩벽법진행AECⅡ적원대배양급순화,장AECⅡ수궤분위고양조급공기조.공기조세포치우5％CO2부육상중배양,고양조세포치우5％CO2 ±95％O2적혼합기체부육상중배양.재배양적2、4、6、8d분별관찰세포적생장정황,검측세포활력、세포주기、세포조망정황.시간점여배양조간적교호작용채용석인설계적방차분석.2조자료비교채용독립양본t검험,다조자료비교채용단인소방차분석,조간량량비교채용Bonferroni방법. 결과 (1)세포생장상황:고양조배양2、4、6화8d시세포수수배양시간연장축점감소,분별위(7.29±0.43)×105/ml、(2.68±0.37)×105/ml、(0.23±0.10)×105/ml화(0.00±0.00)×105/ml,저우상응시간점공기조[분별위(10.41±0.24)×l05/ml、(27.90±1.91)×105/ml、(27.12±0.85)×105/ml화(26.29±1.59)×105/ml],차이균유통계학의의(t분별위10.992、38.912、94.166화49.696,P균=0.000).(2)세포활력:고양조재2、4、6d시세포활력축점하강,활세포솔분별위(79.00±0.71)％、(52.80±1.14)％화(31.60±1.52)％,균저우상응시간점공기조,분별위(97.00±0.71)％、(97.20±0.84)％화(95.00±0.71)％,차이균유통계학의의(t분별위31.213、70.519화84.722,P균=0.000).(3)세포주기변화:고양조G1기화S기세포백분솔재4화6d시여2d시비교균유승고,차고우공기조,차이균유통계학의의(P＜0.05).(4)세포조망변화:고양조Annexin-V+/PI아군재4d시소점비례최고,위(23.89±0.52)％,고우2d화6d시[(21.32±0.43)％화(1.47±0.61)％]；Annexin-V+/PI+아군세포소점비례6d시최고,위(53.92±1.64)％,기차시4d시[(45.03±1.01)％],최저시2d시[(12.17±0.60)％]；고양조각시간점Annexin-V+/PI-화Annexin-V+/PI+아군소점비례여대조조비교,차이균유통계학의의(P＜0.05). 결론 고양대AECⅡ적생장、활력、증식능력、세포주기구유명현적억제작용,대세포조망유촉진작용.
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)×105/ml,(2.68±0.37)×105/ml,(0.23±0.10)×105/ml and (0.00±0.00) × 105/ml],and lower than those in air group [(10.41 ± 0.24) × 105/ml,(27.90±1.91) × 105/ml,(27.12±0.85) ×105/ml and (26.29±1.59) × 105/ml](t=10.992,38.912,94.166and 49.696,P=0.000 respectively). (2) Cell activity:the living cells ratio in hyperoxia group at 2 d[(79.00±0.71) ％],4 d [(52.80±1.14)％] and 6 d [(31.60±1.52)％] was lower than those [(97.00±0.71)％,(97.20±0.84)％ and (95.00±0.71)％] ir 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.
