己酮可可碱干预氧化应激 线粒体机制在氯气致大鼠急性肺损伤中的作用  

Protective effects of pentoxifylline against chlorine-induced acute lung injury by intervening oxidative stress-mitochondria mechanism in rats

作　　者：刘萌萌 赵晨茜 刘姗姗 刘江正 孔德钦 刘瑞 郭鹏[1] 海春旭 张晓迪 LIU Mengmeng;ZHAO Chenqian;LIU Shanshan;LIU Jiangzheng;KONG Deqin;LIU Rui;GUO Peng;HAI Chunxu;ZHANG Xiaodi(Department of Health Service,Logistics University of People s Armed Police Force,Tianjin 300309,China;Shaanxi Provincial Key Laboratory of Free Radical Biology and Medicine,Ministry-of-Education Key Laboratory of Hazard Assessment and Control in Special Operational Environment,Department of Military Toxicology and Chemical Preventive Medicine,School of Military Preventive Medicine,Air Force Medical University,Xi an 710032,China)

机构地区：[1]武警后勤学院卫生勤务系,天津300309 [2]空军军医大学军事预防医学系军事毒理学与防化医学教研室,陕西省自由基生物学与医学重点实验室,特殊作业环境危害评估与防治教育部重点实验室,陕西西安710032

出　　处：《空军军医大学学报》2022年第6期690-695,共6页Journal of Air Force Medical University

基　　金：国家自然科学基金(32100996);军事医学创新工程专项(16CXZ021);武警后勤学院基础研究项目(WHJ202015)。

摘　　要：目的观察氯气致大鼠急性肺损伤(ALI)过程中己酮可可碱(PTX)的干预作用,探讨其可能的保护机制。方法将24只雄性SD大鼠随机分为对照组、氯气暴露组、PTX干预组(氯气暴露)、单纯PTX组(空气暴露),每组6只。PTX干预组和单纯PTX组大鼠在氯气暴露前30 min和染毒后15 min分两次分别给予100 mg/kg PTX灌胃。染毒剂量为1200 mg/m^(3),时长5 min,对照组给予同等条件下的空气暴露,氯气暴露组给予等量生理盐水灌胃。HE染色后观察肺损伤程度;透射电镜下观察肺组织线粒体超微结构;二辛可宁酸蛋白定量法测定血浆和支气管肺泡灌注液(BALF)中蛋白含量并计算肺通透指数;采用Mito SOX探针检测肺组织活性氧(ROS)的含量;试剂盒检测BALF中丙二醛(MDA)、氧化型谷胱甘肽(GSSG)和还原型谷胱甘肽(GSH)的含量以及超氧化物歧化酶(SOD)的活力;定磷法检测肺组织Na^(+),K^(+)-ATPase和Ca^(2+),Mg^(2+)-ATPase的活力;Western blotting法检测肺组织中过氧化氢酶(CAT)和线粒体外膜转位酶20(TOM 20)蛋白表达水平。结果与对照组相比,氯气暴露后3 h,肺泡结构异常,线粒体肿胀明显,肺组织ROS水平升高;BALF中MDA水平、GSSG水平、SOD活力、TOM 20蛋白表达水平、Na^(+),K^(+)-ATPase和Ca^(2+),Mg^(2+)-ATPase活力升高,GSH含量和GSH/GSSG比值、CAT蛋白表达水平下降,且差异有统计学意义(P<0.05);与氯气暴露组相比,PTX干预后,可下调MDA水平、GSSG水平、TOM 20蛋白表达水平、Na^(+),K^(+)-ATPase和Ca^(2+),Mg^(2+)-ATPase活力(P<0.05),上调GSH含量、GSH/GSSG比值和CAT蛋白表达水平(P<0.05)。结论PTX可能通过影响线粒体数量和功能,减轻肺内氧化损伤,在氯气暴露引起的ALI的过程中起保护作用。Objective To investigate the protective effect of pentoxifylline(PTX)on chlorine-induced acute lung injury(ALI)and explore its potential mechanism.Methods Twenty-four healthy male Sprague-Dawley(SD)rats were randomly divided into 4 groups:control group,chlorine group,chlorine+PTX group and PTX group,with 6 rats in each group.Chlorine+PTX and PTX groups were administrated with PTX(100 mg/kg)at 30 min before chlorine exposure and 15 min after chlorine exposure(1200 mg/m^(3)×5 min).Chlorine and control groups were given air exposure or intragastric administration of normal saline under the same conditions.HE staining was used to observe the degree of lung injury.Transmission electron microscope was used to observe ultrastructural changes of mitochondria in lung tissues.The protein content in plasma and bronchoalveolar lavage fluid(BALF)was determined by bicinchoninic acid and lung permeability index was calculated.The level of pulmonary reactive oxygen species(ROS)was evaluated by Mito SOX probe.The levels of malondialdehyde(MDA),oxidized glutathione(GSSG),L-glutathione(GSH)and the activity of superoxide dismutase(SOD)in BALF were detected by the kit.The activities of Na^(+),K^(+)-ATPase and Ca 2+,Mg 2+-ATPase in lung tissues were detected by phosphate method.Western blotting was performed for the expressions of catalase(CAT)and translocase of the outer mitochondrial membrane 20(TOM 20)in lung tissues.Results Compared with the control group,3 h after chlorine exposure,alveolar structure was abnormal,mitochondrial swelling was obvious,and ROS level in lung tissues increased.The levels of MDA,GSSG,and TOM 20 protein expression and activities of SOD,Na^(+),K^(+)-ATPase and Ca 2+,Mg 2+-ATPase in BALF increased,while GSH level,GSH/GSSG ratio and CAT protein expression level decreased,with statistical significance(P<0.05).Compared with chlorine group,PTX intervention could down-regulate levels of MDA,GSSG,TOM 20 protein expression and activities of Na^(+),K^(+)-ATPase and Ca 2+,Mg 2+-ATPase(P<0.05),and up-regulate GSH le

关 键 词：氯气 己酮可可碱 急性肺损伤 线粒体 氧化损伤 

分 类 号：R965[医药卫生—药理学]