细胞色素P450表氧化酶2J2基因过表达通过抗炎减轻大鼠肺缺血再灌注损伤  

Cytochrome P450 epoxygenase 2J2 overexpression protects against lung ischemia-reperfusion injury in rats via anti-inflammation

作　　者：陈文树 郑冠英[2] 李武锦 平伟 付向宁[3] 王家宁 Chen Wenshu;Zheng Guanying;Li Wujin;Ping Wei;Fu Xiangning;Wang Jianing(Department of Thoracic Surgery,Fujian Provincial Hospital,Provincial Clinic College of Fujian Medical University,Fuzhou 350001,China;Department of Respiratory Medicine,Fujian Provincial Hospital,Provincial Clinic College of Fujian Medical University,Fuzhou 350001,China;Department of Thoracic Surgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]福建医科大学省立临床医学院、福建省立医院胸外科,福州350001 [2]福建医科大学省立临床医学院、福建省立医院呼吸内科,福州350001 [3]华中科技大学同济医学院附属同济医院胸外科,武汉430030

出　　处：《中华实验外科杂志》2019年第5期880-883,共4页Chinese Journal of Experimental Surgery

基　　金：福建省自然科学基金(2015J05142);福建省卫生厅资助项目(2014-2-2).

摘　　要：目的探讨细胞色素P450表氧化酶2J2(CYP2J2)基因过表达对大鼠肺缺血再灌注损伤的作用及其机制。方法通过pcDNA3.1质粒将CYP2J2基因转染至SD大鼠,增加大鼠肺组织中CYP2J2蛋白及循环中相应代谢产物的表达。实验动物随机分为5组:空白对照组(Blank)、单纯开胸组(Sham)、肺缺血再灌注组(IR)、肺缺血再灌注+pcDNA3.1-GFP基因转染组(IR+GFP)、肺缺血再灌注+pcDNA3.1-CYP2J2基因转染组(IR+CYP2J2)。并通过阻断左侧肺门1 h,然后恢复通气和血流灌注2 h来构建大鼠急性肺缺血再灌注损伤模型。实验结束后,处死大鼠,评估肺损伤严重程度,并探讨机制。结果与对照组相比,缺血再灌注处理可导致大鼠急性肺损伤,包括增加肺重/体重的比值(LW/BW)、肺湿重/干重比值(W/D)、支气管肺泡灌洗液蛋白浓度(BALF)、以及大鼠血浆IL-1β、IL-8、TNF-α、sP-selectin、sE-selectin等炎症介质水平,降低抗炎细胞因子IL-10水平,而CYP2J2基因转染可减轻大鼠肺缺血再灌注损伤[LW/BW:(4.720±0.756)×10^-3比(4.761±0.905)×10-3比(7.331±0.484)×10^-3比(7.434±0.702)×10^-3比(6.088±0.405)×10^-3,P<0.01;W/D:3.762±0.537比4.084±0.517比7.726±0.895比7.941±1.015比5.633±0.543,P<0.01;BALF:(664.827±219.547)mg/L比(637.381±83.860)mg/L比(2 138.503±81.156)mg/L比(2 130.880±473.470)mg/L比(1 208.030±256.255)mg/L,P<0.01;IL-1β:(28.100±2.930)ng/L比(30.076±5.550)ng/L比(68.532±5.587)ng/L比(66.935±6.469)ng/L比(51.450±6.300)ng/L,P<0.01;IL-8:(38.949±6.441)ng/L比(45.212±5.814)ng/L比(64.661±11.187)ng/L比(64.136±8.634)ng/L比(52.702±6.392)ng/L,P<0.01;TNF-α:(45.359±6.531)ng/L比(98.797±6.610)ng/L比(188.368±16.373)ng/L比(189.677±19.264)ng/L比(121.861±10.994)ng/L,P<0.01;sP-selectin:(4.855±3.032)μg/L比(16.670±8.177)μg/L比(59.428±11.792)μg/L比(53.845±6.925)μg/L比(40.017±8.545)μg/L,P<0.01;sE-selectin:(499.907±151.751)ng/L比(985.640±144.445)ng/L比(1 826.863±370.931)ng/L比(1 684.189±330.473)ng/L比(1 313.309�Objective To discuss the effect and mechanism of cytochrome P450 epoxygenase 2J2(CYP2J2)overexpression on lung ischemia-reperfusion injury in rats.Methods CYP2J2 gene was transfected by pcDNA3.1 plasmid into Sprague Dawley(SD)rats,which increased the expression of CYP2J2 protein in lung tissue and the circulating level of corresponding metabolites.Then,a rat model of acute lung ischemia-reperfusion injury was structured by clamping the left lung hilar for 1 hour,followed by reperfusion for 2 hours.Animals were randomly divided into 5 groups:Blank(blank control group),Sham(sham group),IR(lung ischemia/reperfusion group),IR+GFP(lung ischemia/reperfusion and pcDNA3.1-GFP gene transfection group),and IR+CYP2J2(lung ischemia/reperfusion and pcDNA3.1-CYP2J2 gene transfection group).Animals were sacrificed at the end of the experiment to evaluate lung injury and explore the mechanism.Results Lung ischemia-reperfusion led to acute lung injury,including increased lung weight/body weight(LW/BW)and lung wet/dry ratios(W/D),increased protein concentration in bronchoalveolar lavage fluid(BALF),increased level of inflammatory mediators in serum(including IL-1β,IL-8,TNF-α,sP-selectin,sE-selectin),and decreased level of IL-10(a anti-inflammatory mediator).However,these effects were significantly relieved by CYP2J2 gene delivery[LW/BW:(4.720±0.756)×10^-3 vs.(4.761±0.905)×10^-3 vs.(7.331±0.484)×10^-3 vs.(7.434±0.702)×10^-3 vs.(6.088±0.405)×10^-3,P<0.01;W/D:3.762±0.537 vs.4.084±0.517 vs.7.726±0.895 vs.7.941±1.015 vs.5.633±0.543,P<0.01;BALF:(664.827±219.547)mg/L vs.(637.381±83.860)mg/L vs.(2 138.503±81.156)mg/L vs.(2 130.880±473.470)mg/L vs.(1 208.030±256.255)mg/L,P<0.01;IL-1β:(28.100±2.930)ng/L vs.(30.076±5.550)ng/L vs.(68.532±5.587)ng/L vs.(66.935±6.469)ng/L vs.(51.450±6.300)ng/L,P<0.01;IL-8:(38.949±6.441)ng/L vs.(45.212±5.814)ng/L vs.(64.661±11.187)ng/L vs.(64.136±8.634)ng/L vs.(52.702±6.392)ng/L,P<0.01;TNF-α:(45.359±6.531)ng/L vs.(98.797±6.610)ng/L vs.(188.368±16.373)ng/L vs.(189.677±19.264)

关 键 词：细胞色素P450表氧化酶2J2基因过表达 肺缺血再灌注损伤 急性肺损伤 

分 类 号：R563[医药卫生—呼吸系统]