异氟烷预处理或后处理对大鼠局灶性脑缺血/再灌注损伤的影响  

The effects of preconditioning and postconditioning with isoflurane on focal cerebral ischemi/reperfusion injury in rats

作　　者：李国福[1] 贾佳[1] 符加红[1] 汪海源[1] 吉凯强[1] 臧彬[1] 

机构地区：[1]中国医科大学附属盛京医院重症医学科,辽宁沈阳110004

出　　处：《中华危重病急救医学》2014年第6期431-435,共5页Chinese Critical Care Medicine

基　　金：辽宁省科技厅科学技术计划项目（2010225034）

摘　　要：目的 探讨异氟烷预处理或后处理对大鼠局灶性脑缺血/再灌注（I/R）时炎症因子、脂质过氧化等相关指标的影响.方法 将32只SD大鼠按随机数字表法分为对照组、模型组、异氟烷预处理组及异氟烷后处理组4组,每组8只.对照组不给予异氟烷与脑缺血处理;模型组大鼠接受90 min右侧大脑中动脉闭塞（MCAO）;异氟烷预处理组大鼠在MCAO前24 h给予2％异氟烷30 min;异氟烷后处理组大鼠在MCAO后,于再灌注开始时给予2％异氟烷60 min.操作结束24 h后取心脏血,检测血清炎症因子白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）含量及脂质过氧化相关指标丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性.采用逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹试验（Westen Blot）检测右侧脑组织基质金属蛋白酶（MMP-2、MMP-9）及紧密连接蛋白闭合蛋白5（Claudin-5）、咬合蛋白（Occludin）的mRNA和蛋白表达.结果 与对照组相比,模型组大鼠血清IL-1β（ng/L）、TNF-α（ng/L）及MDA（μmol/L）含量均显著增加,SOD（U/L）活性则显著下降（IL-1β：76.81±11.14比52.43±8.86,TNF-α：64.93±10.81比33.64±7.94,MDA：8.63±1.42比4.14±0.98,SOD：0.95±0.21比2.36±0.80,均P＜0.05）;与模型组比较,经异氟烷预处理或后处理后,大鼠血清IL-1β、TNF-α及MDA含量显著下降,SOD活性则显著回升（IL-1β：54.37±9.06、56.82±8.67比76.81±11.14,TNF-α：43.72±6.16、39.49±9.34比64.93±10.81,MDA：5.65 ±0.83、5.82±0.78比8.63±1.42,SOD：1.64±0.47、1.71±0.52比0.95±0.21,均P＜0.05）.同时,I/R可导致受损脑组织MMP表达升高,紧密连接蛋白表达降低;与模型组比较,经过异氟烷预处理或后处理后,大鼠受损脑组织中MMP-2与MMP-9的mRNA及蛋白表达均明显下降（MMP-2 mRNA：1.25±0.08、1.32±0.12比2.48±0.26,MMP-2蛋白：1.56±0.09、1.50±0.08比2.12±0.11; MMP-9 mRNA：1.26 ±0.13、1.20±0.12比2.74±0.28,MMP-9蛋白�Objective To investigate the effects of preconditioning and postconditioning with isoflurane on pro-inflammatory cytokines and lipid peroxidation in focal cerebral ischemic/reperfusion （I/R） injury in rats.Methods Thirty-two Sprague-Dawley （SD） rats were randomly divided into four groups：control group,model group,isoflurane preconditioning group and isoflurane postconditioning group,with 8 rats in each group.Rats in control group did not receive any challenge.In rats of model group right middle cerebral artery occlusion （MCAO） was conducted for 90 minutes.Rats in isoflurane preconditioning group received 2％ isoflurane exposure for 30 minutes 24 hours before MCAO for 90 minutes.Rats in isoflurane postconditioning group were given 60-minute 2％ isoflurane exposure after reperfusion of right MCAO.Twenty-four hours after the procedure,all rats were anesthetized with isoflurane,and blood sample taken from the heart was centrifuged,and the pro-inflammatory cytokines,including interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α）,and lipid peroxidation products such as malonaldehyde （MDA） and superoxide dismutase （SOD） were determined.The mRNA and protein expression levels of matrix metalloproteinase （MMP-2,MMP-9）,tight junction protein Calaudin-5 and Occludin were determined by reverse transcription-polymerase chain reaction （RT-PCR） and Western Blot.Results Compared with control group,serum levels of IL-1 β （ng/L）,TNF-α （ng/L） and MDA （μmol/L） were elevated and activity of SOD （U/L） decreased in rats of model group （IL-1β：76.81 ± 11.14 vs.52.43 ± 8.86,TNF-α：64.93 ± 10.81 vs.33.64 ± 7.94,MDA：8.63 ± 1.42 vs.4.14 ± 0.98,SOD：0.95 ± 0.21 vs.2.36 ± 0.80,all P＜0.05）.After isoflurane preconditioning and postconditioning,compared with model group,the levels of IL-1 β,TNF-α and MDA were lowered,while activity of SOD was increased （IL-1 β：54.37 ± 9.06,56.82 ± 8.67 vs.76.81 ± 1 1.14,TNF-α：43.72 ± 6.16,39.49 ± 9.34 vs.64.93

关 键 词：异氟烷 预处理 后处理 缺血/再灌注损伤 脑 

分 类 号：R965[医药卫生—药理学]