乌司他丁对硫化氢急性中毒大鼠肺组织氧化应激损伤的干预作用及机制研究  

Effect of ulinastatin on oxidative stress in the lung tissue of acute H2 S-intoxicated rats

作　　者：葛赟[1] 余毅娟[1] 郑嘉奕[1] 刘瑶[1] 邱俏檬[1] 洪广亮[1] 胡国新[1] 李萌芳[1] 卢中秋[1] 

机构地区：[1]温州医学院附属第一医院急诊科,浙江省温州325000

出　　处：《中华急诊医学杂志》2012年第2期164-170,共7页Chinese Journal of Emergency Medicine

基　　金：基金项目：浙江省医学扶植重点建设学科计划（07-F04）;浙江省医药卫生科技计划（20088126）

摘　　要：目的 观察硫化氢（hydrogen sulfide,H2S）急性中毒大鼠肺组织血红素氧合酶-1（heme oxygenase 1,HO-1）、醌氧化还原酶-1（NAD（P）H：quinine oxidoreductase 1,NQO-1）和核转录因子红系相关因子-2（nuclear factor E2-related factor 2,Nrf2）的动态变化及乌司他丁（ulinastatin,UTI对其的影响.方法 将清洁级SD大鼠96只随机（随机数字法）分为健康对照组（NS组,n=8）,UTI对照组（UTI组,n=8）,H2S染毒模型组（H2S组,n=40,采用在染毒柜内暴露吸入体积分数200×10-6的H2S1h,构建H2S急性染毒模型）和UTI干预组（H2S＋UTI组,n=40,建立模型后立即腹腔注射UTI 10万U/kg）.后两组大鼠于染毒后2、6、12、24、48 h时点麻醉后活杀,留取肺标本.观察大鼠行为学变化,ELISA法测定肺组织中HO-1和NQO-1活力的动态变化；RT-PCR法检测肺组织HO-1、NQO-1和Nrf2 mRNA表达,Western blot法检测Nrf2蛋白表达.观察肺组织病理学改变并行肺损伤评分.结果 与NS组比较,H2S组在染毒后2、6、12h时点HO-1活力和mRNA表达均明显升高（P＜0.01）,其中2h呈高峰；与同时间点H2S组比较,H2S＋ UTI组染毒后6、12、24、48 h时点HO-1活力和mRNA表达显著升高（P＜0.01）.与 、NS组比较,H2S组在染毒后2、6、12、24 h时点NQO-1活力和mRNA表达均明显升高（P＜0.01）其中2h呈高峰；与同时间点H2S组比较,H2S＋UTI组染毒后6、12、24、48 h时点NQO-1活力和mRNA表达显著升高（P＜0.01）.与NS组比较,H2S组在染毒后2、6、12 h时点Nrf2mRNA和蛋白表达均明显升高（P＜0.01或P＜0.05）,其中2h呈高峰；与同时间点H2S组比较,H2S＋UTI组染毒后6、12、24、48 h时点Nrf2 mRNA和蛋白表达显著升高（P＜0.01）.光镜下,H2S组在染毒后24 h大鼠肺组织损害明显；H2S＋UTI组大鼠肺损伤较H2S组有所减轻.在染毒后12、24、48 h时点H2S＋UTI组的肺损伤评分明显低于H2S组（P＜0.01）.结论 HO-1、NQO-1和Nrf2参与了H2S中毒急性肺损伤的病理生Objective To observe the dynamic changes of heme oxygenase 1,NAD（P）H：quinine oxidoreductase 1 and Nuclear factor-E2-related factor 2 in the lung tissue of acute H2S-intoxicated rats and intervention effects of ulinastratin（UTI）.Methods A total of 96 SD rats of clean grade were divided randomly（random number）into four groups：normal control group（NS group,n =8）,UTI control group（UTI group,n =8）,H2S-intoxicated model group（H2S group,n =40,rats were exposed to H2S（200 × 10-6）for 1 h to establish the H2S-intoxicated model）and UTI treatment group（H2S ＋UTI group,n =40,rats were intraperitoneal injected with the dose of UTI 105 U/kg）.H2S group and H2S ＋ UTI group were sacrificed 2,6,12,24 and 48 h after modeling.The activity and mRNA expression of HO-1 and NQO-1 in the lung tissue were measured by ELISA and RT-PCR methods,and the expression of Nrf2 mRNA and protein in the lung tissue was detected by RT-PCR and Western Blot methods.Pathological changes of lung tissue were observed by lightmicroscope and the lung injury score was used to evaluate inhalation injury.Results The pulmonary HO-1 activity and mRNA expression in rats of H2S group at 2,6,12 h（P 〈 0.01）after intoxication were markedly increased than that in NS group：In comparison with H2S group,the pulmonary HO-1 activity and mRNA expression increased at 6,12,24,48 h（P 〈0.01）.The pulmonary NQO-1 activity and mRNA expression in rats of H2S group at 2,6,12,24 h（P〈 0.01）after intoxication were markedly increased than that in NS group; In comparison with H2S group,the pulmonary NQO-1 activity and mRNA expression increased at 6,12,24,48 h（P 〈 0.01）.The pulmonary Nrf2 mRNA and protein expression in rats of H2S group at 2,6,12 h（P 〈0.01 or P 〈0.05）after modeling were markedly increased than that in NS group and reached peak 2 hour after modeling; In comparison with H2S group,the pulmonary Nrf2 mRNA and protein expression increased at 6,12,24,48 h（P 〈0.01）.At 24 h after modeling,the degree of

关 键 词：硫化氢 乌司他丁 急性肺损伤 氧化应激 核转录因子红系相关因子-2 血红素氧合酶-1 醌氧化还原酶-1 大鼠 

分 类 号：R595.1[医药卫生—内科学]