野生型p53诱导细胞凋亡在海水淹溺型大鼠肺损伤中的作用  

Effect of apoptosis induced by wild - type p53 on the acute lung injury of rats challenged with seawater drowning

作　　者：刘庆晴 罗颖[2] 李聪聪[1] 薄丽艳[1] 刘伟[1] 徐敦全[2] 陈向军[1] 李燕燕[1] 金发光[1] 

机构地区：[1]第四军医大学唐都医院呼吸与危重症医学科,陕西西安710038 [2]第四军医大学基础部病理与病理生理学教研室,陕西西安710032

出　　处：《中国急救医学》2014年第1期48-52,I0003,共6页Chinese Journal of Critical Care Medicine

基　　金：军队“十一五”攻关基金资助项目（08G-102）;陕西省攻关基金资助项目（2008K14-08）

摘　　要：目的观察野生型p53（wtp53）基因在海水淹溺型肺损伤大鼠肺组织中的表达，探讨细胞凋亡参与肺损伤的发病机制。方法32只SD大鼠随机分为正常对照组（NG）及海水2、4、8h组，每组8只。三个海水处理组大鼠气管内注入4mL／kg海水，分别作用2h、4h和8h；对照组除不吸入海水外，其余处理同海水淹溺组。于各时间点放血处死大鼠，测肺湿／干质量比（W／D），HE染色观察肺组织病理形态改变，TUNEL法检测肺组织细胞凋亡，RT—PCR检测叭p53及细胞色素C（cytochrome—C，Cyt—C）mRNA的表达，Westernblotting分析wtp53和Cyt—C及活化caspase-3的蛋白表达。结果与正常对照组比较，各海水组W／D比值升高（P〈0．01），肺组织细胞凋亡率明显增加（P〈0．05，P〈0．01），肺组织wtp53及Cyt—CmRNA表达均明显升高（P〈0．01），wtp53和Cyt—C及活化caspase-3蛋白表达随时间增加明显上调（P〈0．01，P〈0．05）。结论海水诱发大鼠淹溺型肺损伤和肺组织细胞凋亡，野生型p53、Cyt—C和活化的caspase-3的表达增加，野生型p53在海水淹溺肺损伤细胞凋亡发生发展中起重要作用。Objective To observe the expression of wild - type p53 （ wtp53 ） gene in lung tissues of the acute lung injury model induced by seawater drowning （ SWD -ALI） in rats and to explore the mechanism of apoptosis in the pathogenesis of lung injury. Methods 32 male Sprague - Dawley rats were randomly divided into 4 groups ： normal control group （ NG, n = 8 ）, 2 h, 4 h and 8 h seawater group （n = 8）. Seawater （4 mL/kg） was instilled into both lungs of the rats and then the rats were killed at 2 h, 4 h and 8 h in seawater groups, respectively. Lung wet/dry weight （W/D） was calculated. Pulmonary histological changes were investigated by hE. The cell apoptosis rate in rat lung tissues was detected by TUNEL. The expression of wtp53 and Cyt - c mRNA were performed by RT - PCR. The protein expression of wtpS, Cyt - c and active caspase - 3 was analyzed by Western blotting. Results Compared with NG group, acute lung injury was induced significantly after seawater injection. The W/D ratio of seawater group was elevated （P 〈 0.01 ）. The cell apoptosis rate was prominently increased in seawater groups （2 h, P 〈0.05; 4 h and 8 h, P 〈0.01）. The expression of wtp53 and Cyt - c mRNA was remarkably higher in seawater groups than that in NG group （ P 〈 0.01 ）, and the protein expression of wtp53, Cyt - c and active caspase - 3 was obviously up - regulated in seawater groups （P 〈 0.01, P 〈 0.05）. Conclusion Seawater could induce the rat lung injury and lung tissue cell apoptosis and enhance the expression of wtp53, Cyt -c and active caspase -3, and wtp53 plays a key role in the apoptosis of SWD - ALL

关 键 词：海水淹溺 急性肺损伤 凋亡 野生型r63(wtp53) 细胞色素c(Cyt—c) 

分 类 号：R730.5[医药卫生—肿瘤]