七氟烷后处理对大鼠脑缺血再灌注损伤保护作用的研究  

Protective effect of sevoflurane postconditioning against cerebral ischemia/reperfusion injury in rats

作　　者：毛艺锟 王士雷[1] 吴秀云[1] 赵芹[1] 李瑜[1] MAO Yikun;WANG Shilei;WU Xiuyun;ZHAO Qin;LI Yu(Department of Anesthesiology,The Affiliated Hospital of Qingdao University,Qingdao 266555,China)

机构地区：[1]青岛大学附属医院麻醉科,山东青岛266555

出　　处：《精准医学杂志》2024年第2期125-129,133,共6页Journal of Precision Medicine

基　　金：国家自然科学基金面上项目(81771415)。

摘　　要：目的探讨七氟烷后处理对大鼠脑缺血再灌注(I/R)损伤的保护作用及其机制。方法选择SPF级成年健康雄性SD大鼠80只,随机分为假手术组(S组)、脑缺血再灌注组(I/R组)、脑I/R+七氟烷后处理组(ISP组)、脑I/R+七氟烷后处理+核因子E2相关因子2(Nrf2)抑制剂组(ISPB组),每组20只。除S组外,其余组大鼠均用线栓法闭塞大脑中动脉2 h并再灌注24 h的方法制备脑I/R损伤大鼠模型(S组大鼠只在大脑中动脉下穿线不结扎)。ISP组大鼠于再灌注即刻吸入3%七氟烷30 min,ISPB组在缺血前30 min腹腔注射Nrf2抑制剂鸦胆子苦醇(2 mg/kg),其余处理同ISP组。建模成功后通过神经功能评分评估各组大鼠神经功能损害程度。随后获取大鼠左心室血及脑组织病理切片,以2,3,5-氯化三苯基四氮唑(TTC)染色测定各组大鼠脑梗死体积百分比,采用酶联免疫吸附试验检测大鼠血清中炎症因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和氧化应激相关因子丙二醛(MDA)及超氧化物歧化酶(SOD)水平,采用免疫印迹实验检测大鼠脑组织中凋亡相关蛋白B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关联x(Bax)、半胱胺酸天冬氨酸蛋白酶-3(Caspase-3)及Nrf2信号通路相关蛋白Nrf2和血红素加氧酶1(HO-1)表达,采用免疫荧光实验检测大鼠脑组织细胞核内外Nrf2表达。结果与I/R组相比,ISP组大鼠神经功能缺损评分、脑梗死体积百分比,以及血清IL-1β、TNF-α、MDA水平和脑组织总蛋白Bax、Caspase-3水平均下降(t=5.76~18.39,P<0.05);血清中SOD水平及脑组织总蛋白Nrf2、HO-1、Bcl-2水平均升高(t=5.73~14.08,P<0.05),Nrf2免疫荧光强度增强。与ISP组相比,ISPB组神经功能缺损评分、脑梗死体积百分比,以及血清中IL-1β、TNF-α、MDA水平和脑组织总蛋白Bax、Caspase-3水平均升高(t=3.06~8.19,P<0.05);血清中SOD水平和脑组织总蛋白Nrf2、HO-1、Bcl-2水平均下降(t=2.67~9.01,P<0.05),Nrf2免疫荧光强度减弱。结论�Objective To investigate the protective effect of sevoflurane postconditioning against cerebral ischemia/reperfusion(I/R)injury in rats and its mechanism of action.Methods A total of 80 specific pathogen-free healthy adult male Sprague-Dawley rats were selected and randomly divided into sham-operation group(S group),cerebral I/R group(I/R group),cerebral I/R+sevoflurane postconditioning group(ISP group),and cerebral I/R+sevoflurane postconditioning+nuclear factor ery-throid 2-related factor 2(Nrf2)inhibitor group(ISPB group),with 20 rats in each group.All rats except those in the S group were used to establish a rat model of cerebral I/R injury using the suture method for occlusion of the middle cerebral artery for 2 h,followed by reperfusion for 24 h,and for the rats in the S group,threading was performed below the middle cerebral artery without ligation.The rats in the ISP group were given inhalation of 3%sevoflurane immediately after reperfusion for 30 min,and those in the ISPB group were given intraperitoneal injection of the Nrf2 inhibitor brusatol(2 mg/kg)at 30 min before ischemia in addition to the treatment in the ISP group.After successful modeling,neurological deficit score was used to eval-uate the degree of neurological impairment.Left ventricular blood samples and pathological sections of brain tissue were obtained,and 2,3,5-triphenyltetrazolium chloride staining was used to determine the percentage of cerebral infarct volume;enzyme-linked immunosorbent assay was used to measure the serum levels of inflammatory factors(interleukin-1βand tumor necrosis factor-α)and oxidative stress-related factors(malondialdehyde and superoxide dismutase);Western blotting was used to mea-sure the expression of apoptosis-related proteins(B-cell lymphoma-2,Bcl-2 related x,and Caspase-3)and Nrf2 signaling pathway-related proteins(Nrf2 and heme oxygenase-1)in brain tissue;immunofluorescence assay was used to measure the expression of Nrf2 inside and outside the nucleus of brain tissue cells.Results Compared with the I/R

关 键 词：七氟烷 缺血后处理 脑缺血 再灌注损伤 NF-E2相关因子2 信号传导 大鼠 Sprague-Dawley 

分 类 号：R619.9[医药卫生—外科学] R743.31[医药卫生—临床医学]