乳化异氟醚后处理对大鼠心肌缺血再灌注时Nrf2-ARE信号通路的影响：离体实验  

Effect of emulsified isoflurane postconditioning on Nrf2-ARE signaling pathway during myocardial ischemia-reperfusion in rats in vitro

作　　者：杜文娟[1,2] 王海英[1] 李小娟[1,3] 陈伟[1] 徐鹏[1] 喻田[1] 

机构地区：[1]遵义医学院附属医院麻醉科、贵州省麻醉与器官保护基础研究重点实验室,563000 [2]山西长治市人民医院正骨科 [3]湘潭市中心医院麻醉科

出　　处：《中华麻醉学杂志》2015年第5期632-636,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（30960366）

摘　　要：目的：评价乳化异氟醚后处理对大鼠缺血再灌注时核转录因子 NF-E2相关因子2（ Nrf2）-抗氧化反应元件（ ARE）信号通路的影响。方法健康雄性SD大鼠，体重250～300 g，4～5月龄，采用Langendorff灌注装置建立大鼠离体心脏灌注模型。取模型制备成功的心脏32个，采用随机数字表法分为4组（ n＝8）：对照组（ C组）、缺血再灌注组（ I∕R组）、乳化异氟醚后处理组（ EIP组）和脂肪乳组（ F组）。平衡灌注20 min后，C组继续灌注100 min；I∕R组32℃下缺血40 min，恢复灌注60 min；EIP组32℃下缺血40 min，于再灌注前即刻灌注含乳化异氟醚1．68 mmol∕L的K-H液2 min，继续灌注37℃含氧的K-H液58 min；F组32℃下缺血40 min，于再灌注前即刻灌注含脂肪乳712 mg∕L 的K-H液2 min，继续灌注37℃含氧K-H液58 min。分别于平衡灌注末及再灌注末记录HR、左室发展压（LVDP）、左室舒张末压（LVEDP）和左心室压力最大上升速度（＋dp∕dtmax）。于再灌注末，取左心室心肌组织，观察心肌细胞超微结构，分别采用RT-PCR法和Western blot法检测心肌组织Nrf2、血红素加氧酶-l （HO-1）、醌氧化还原酶1（NQO1）、超氧化物歧化酶1（SOD1）及其mRNA的表达水平。结果与C组比较，再灌注末I∕R组和F组HR、＋dp∕dtmax和LVDP 降低，LVEDP 升高，EIP组LVDP降低，LVEDP升高（ P＜0．05），HR和＋dp∕dtmax差异无统计学意义（ P＞0．05）， I∕R组、EIP组和F组心肌组织Nrf2、HO-1、NQO1和SOD1及其mRNA的表达下调（ P＜0．05）；与I∕R组比较，EIP组和F组再灌注末HR、＋dp∕dtmax和LVDP 升高，LVEDP 降低，EIP 组心肌组织Nrf2、HO-1、NQO1和SOD1及其mRNA的表达上调， F组心肌组织Nrf2、HO-1、NQO1和SOD1的mRNA表达上调，Nrf2和HO-1的表达上调（ P＜0．05），NQO1和SOD1的表达差异无统计学意义（ P＞0．05）；与EIP组比较，F组再灌注末HR、＋dp∕dtmax和LVDP降Objective To evaluate the effect of emulsified isoflurane postconditioning on nuclear factor-E2 related factor 2 （ Nrf2 ）-antioxidant response element （ ARE ） signaling pathway during myocardial ischemia-reperfusion （ I∕R ） in rats in vitro. Methods Healthy male Sprague-Dawley rats, aged 4-5 months, weighing 250-300 g, were heparinized and anesthetized with intraperitoneal 1% amobarbital sodium 40 mg∕kg. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution. Thirty-two isolated rat hearts were randomly divided into 4 groups （ n=8 each ） using a random number table： control group （group C）, group I∕R, emulsified isoflurane postconditioning group （EIP group） and fat emulsion group （ group F） . After 20 min of equilibration, group C was continuously perfused with K-H solusion for 100 min. Group I∕R underwent 40 min of ischemia at 32 ℃, followed by reperfusion for 60 min. In EIP and F groups, after undergoing 40 min of global ischemia, the isolated hearts were perfused for 2 min with K-H solution containing 1.68 mmol∕L emulsified isoflurane and 712 mg∕L intralipid, respectively, starting from the onset of reperfusion, and then were continuously perfused with K-H solution containing oxygen at 37 ℃ for 58 min. Heart rate （HR）, left ventricular developed pressure （LVDP）, left ventricular end-diastolic pressure （ LVEDP ） , and positive maximal pressure of left ventricular increase （＋dp∕dtmax ） were recorded at the end of equilibration and reperfusion. At the end of reperfusion, myocardial specimens were obtained from the left ventricle for examination of the ultrastructure of myocardial cells and for determination of Nrf2, heme oxygenase-1 （ HO-1） , quinone oxidoreductase 1 （ NQO1） , and superoxide dismutase 1 （ SOD1） and mRNA expression using Western blot and real-time PCR. Results Compared with group C, HR, ＋dp∕dtmax and LVDP were significantly decreased, and LVEDP was increased at the end of reperfusion in I

关 键 词：异氟醚 脂肪乳剂 静脉注射用 缺血后处理 心肌再灌注损伤 NF-E2相关因子2 反应元件 

分 类 号：R614[医药卫生—麻醉学]