Nrf2/HO-1通路在电针减轻肢体缺血再灌注诱发兔肺损伤中的作用  

Effect of Nrf2/HO-1 Pathway on Electroacupuncture-induced Reduction of Lung Injury Induced by Limb Ischemia-reperfusion in Rabbits

作　　者：王德强 余剑波[2] 阚永星 宫丽荣[2] WANG De-qiang;YU Jian-bo;KAN Yong-xing(Department of Anesthesiology,Dagang Hospital of Tianjin Binhai New Area,Tianjin (300270),China)

机构地区：[1]天津市滨海新区大港医院麻醉科,天津300270 [2]天津医科大学南开临床学院天津市南开医院麻醉科,天津300100

出　　处：《中国中西医结合外科杂志》2019年第3期271-275,共5页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：天津市滨海新区卫生计生委科技联合攻关项目(2015BWKL003)

摘　　要：目的:观察核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)信号通路在电针减轻肢体缺血再灌注诱发兔肺损伤中的作用。方法:40只健康雄性新西兰大白兔,采用随机数字表法分为假手术组、模型组、电针组和全反式维甲酸组(n=10)。采用夹闭股动脉3h、再灌注4h的方法制备兔肢体缺血再灌注模型;电针组、全反式维甲酸组于模型制备前1~4d(30min/次,1次/d)及模型制备过程中电针足三里穴和肺俞穴,采用疏密波2/15Hz,强度以兔出现轻微肌颤为宜;全反式维甲酸组于模型制备前30min腹腔注射Nrf2抑制剂全反式维甲酸7mg/kg。再灌注4h时采集颈动脉血样,随后处死兔取肺组织,观察病理学结果并行肺损伤评分,计算肺湿/干重(W/D)比值;测定肺组织MDA含量及SOD活性,采用Western blotting法检测肺组织Nrf2总蛋白、Nrf2核蛋白及HO-l蛋白表达水平。结果:模型组、电针组和全反式维甲酸组再灌注4h时肺损伤评分分别为5.43±0.82、4.03±0.56、6.21±0.75,均明显高于假手术组(0.92±0.21,P<0.05);肺组织W/D比值分别为6.42±1.00、4.45±0.68、7.08±1.12,均明显高于假手术组(2.04±0.29,P<0.05);MDA含量分别为(4.82±0.51)nmol/mg、(3.56±0.45)nmol/mg、(5.18±0.47)nmol/mg,均明显高于假手术组(2.10±0.23)nmol/mg,P<0.05;SOD活性分别为(50.6±6.3)U/mg、(63.1±7.7)U/mg、(47.2±5.6)U/mg,均明显低于假手术组(72.6±6.3)U/mg,P<0.05;Nrf2总蛋白、Nrf2核蛋白及HO-l蛋白表达水平均明显高于假手术组(P<0.05)。与模型组比较,电针组再灌注4h时肺损伤评分、肺组织W/D比值、MDA含量均明显降低(P<0.05),SOD活性、Nrf2总蛋白、核蛋白及HO-l蛋白表达水平均明显升高(P<0.05)。与电针组比较,全反式维甲酸组再灌注4h时肺损伤评分、肺组织W/D比值、MDA含量均明显升高(P<0.05),SOD活性、Nrf2总蛋白、核蛋白及HO-l蛋白表达水平均明显降低(P<0.05)。结论:Nrf2/HO-1信号通路激活参与了电针减轻肢体缺血�Objective To observe the effect of nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway on electroacupuncture-induced reduction of lung injury induced by limb ischemiareperfusion in rabbits.Methods Forty New Zealand white rabbits were randomly divided into 4 groups(n=10),such as Sham operation group,limb ischemia-reperfusion group,electrical acupoint stimulation group and all-trans retinoic acid group.Limb ischemia-reperfusion was induced by clamping bilateral femoral arteries for 3 h followed by 4 h reperfusion.Bilateral electroacupuncture stimulation of Zusanli and Feishu acupoints was performed with disperse-dense wave 2/15 Hz for 30 min once daily for 4 d before establishment of the model and during the course of establishment of the model in electrical acupoint stimulation group and all-trans retinoic acid group.At 30 min before establishment of the model,Nrf2 inhibitor all-trans retinoic acid(7 mg/kg)was injected intraperitoneally in all-trans retinoic acid group.The rabbits were sacrificed at 4 h of reperfusion and the lungs were removed for microscopic examination and determination of lung injury scores,wet to dry weight ratio(W/D),malondialdehyde(MDA)contents,superoxide dismutase(SOD)activities,and expression of Nrf2 total protein,nucleoprotein and HO-1 protein by Western blotting.Results The lung W/D ratio after 4 h reperfusion in limb ischemia-reperfusion group,electrical acupoint stimulation group and all-trans retinoic acid group were 6.42±1.00,4.45±0.68 and 7.08±1.12,respectively.The lung injury scores at 4 h in above three groups were 5.43±0.82,4.03±0.56 and 6.21±0.75,respectively.Both lung W/D ratio and lung injury scores in above three groups were significantly higher than those of Sham operation group.MDA contents[(4.82±0.51)nmol/mg,(3.56±0.45)nmol/mg and(5.18±0.47)nmol/mg]in limb ischemia-reperfusion group,electrical acupoint stimulation group and all-trans retinoic acid group were significantly increased compared with Sham operation group[(2.10±0.23)nmol/m

关 键 词：NF-E2相关因子2 血红素氧合酶-1 电刺激疗法 缺血再灌注 肺损伤 

分 类 号：Q95-33[生物学—动物学]