HO-1在血红素结合蛋白减轻大鼠脑缺血再灌注损伤中的作用  

Role of heme oxygenase-1 in hemopexin-induced reduction of cerebral ischemia-reperfusion injury in rats

作　　者：董贝贝[1] 张智申 杨永妍 谢克亮[1] 于泳浩[1] Dong Beibei;Zhang Zhishen;Yang Yongyan;Xie Keliang;Yu Yonghao(Department of Anesthesiology,Tianjin Medical University General Hospital,Tianjin 300052,China;Department of Anesthesiology,Xiamen Medical University Zhongshan Hospital,Xiamen 516211,China)

机构地区：[1]天津医科大学总医院麻醉科,300052 [2]厦门医科大学附属中山医院手术麻醉科,516211

出　　处：《中华麻醉学杂志》2020年第3期342-346,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金青年科学基金(81501013)。

摘　　要：目的评价血红素氧合酶-1(HO-1)在血红素结合蛋白减轻大鼠脑缺血再灌注损伤中的作用。方法雄性SD大鼠180只,7~8周龄,体重250~280 g,采用随机数字表法分为5组(n=36):假手术组(SH组)、缺血再灌注组(I/R组)、溶剂组(V组)、血红素结合蛋白组(HPX组)和血红素结合蛋白+HO-1特异性抑制剂ZnPPIX组(HZ组)。采用线栓法阻塞大鼠大脑中动脉制备局灶性脑缺血再灌注损伤模型,缺血120 min后恢复灌注。再灌注即刻I/R组、V组、HPX组和HZ组侧脑室分别注射0.9%生理盐水10μl、0.1%NaN3溶液10μl、血红素结合蛋白1.86 g/L(用0.1%NaN3溶液稀释至10μl)、血红素结合蛋白1.86 g/L+ZnPPIX 20μmol/L(用0.1%NaN3溶液稀释至10μl)。分别于缺血前1 d和再灌注2 d时采用Morris水迷宫实验测试大鼠认知功能。再灌注7 d时,处死大鼠,取脑组织,检测伊文思蓝(EB)渗透率和含水量;采用Western blot法检测缺血半暗带区脑组织血管内皮钙粘蛋白(VE-cadherin)表达水平;采用RT-PCR检测血管生成素1(Ang1)和Ang2的mRNA表达水平,计算Ang1 mRNA/Ang2 mRNA比值;采用CD31/vWF免疫荧光双标染色法测定缺血半暗带区海马组织新生血管密度。结果与SH组比较,I/R组、V组、HPX组和HZ组再灌注2~7 d时逃避潜伏期延长,目标象限停留时间百分比降低,穿越原平台次数减少,再灌注7 d时脑组织EB渗透率升高,含水量、Ang1 mRNA/Ang2 mRNA比值、VE-cadherin表达水平和新生血管密度降低(P<0.05);与I/R组比较,HPX组再灌注2~7 d时逃避潜伏期缩短,目标象限停留时间百分比升高,穿越原平台次数增加,再灌注7 d时脑组织EB渗透率降低,含水量、Ang1 mRNA/Ang2 mRNA比值、VE-cadherin表达水平和新生血管密度升高(P<0.05),V组和HZ组上述各指标差异无统计学意义(P>0.05)。与HPX组比较,HZ组再灌注2~7 d时逃避潜伏期延长,目标象限停留时间百分比降低,穿越原平台次数减少,再灌注7 d时脑组织EB渗透率升高,含水量、AngObjective To evaluate the role of heme oxygenase-1(HO-1)in hemopexin(HPX)-induced reduction of cerebral ischemia-reperfusion(I/R)injury in rats.Methods One hundred and eighty male Sprague-Dawley rats,aged 7-8 weeks,weighing 250-280 g,were divided into 5 groups(n=36 each)using a random number table method:sham operation group(group SH),I/R group,vehicle group(group V),HPX group,and HPX plus HO-1 specific inhibitor ZnPPIX(HZ group).Local cerebral I/R was produced by middle cerebral artery occlusion for 120 min followed by reperfusion in anesthetized rats.In I/R,V,HPX and HZ groups,0.9%normal saline 10μl,0.1%NaN310μl,1.86 g/L HPX(diluted to 10μl in 0.1%NaN3 solution),and 1.86 g/L+ZnPPIX 20μmol/L(diluted to 10μl in 0.1%NaN3 solution)were injected through the lateral ventricle,respectively,immediately after onset of reperfusion.Morris water maze test was used to detect the cognitive function on day 1 before ischemia and day 2 of reperfusion.Rats were sacrificed,brains were removed and brain tissues were obtained for determination of the permeability ratio of Evans blue(EB),brain water content,expression of VE-cadherin in ischemic penumbra(by Western blot),and expression of angiopoietin-1(Ang1)mRNA and Ang2 mRNA(by real-time polymerase chain reaction).Ang1 mRNA/Ang2 mRNA ratio was calculated.CD31/vWF double labeling immunofluorescence was used to detect the density of neovascularization in hippocampal tissues in the ischemic penumbra.Results Compared with SH group,the escape latency was significantly prolonged at 2-7 days of reperfusion,the percentage of time of staying at the target quadrant was decreased,the frequency of crossing the platform was decreased,the permeability ratio of EB in brain tissues was increased at day 7 of reperfusion,and the brain water content,Ang1 mRNA/Ang2 mRNA ratio,expression of VE-cadherin and density of neovascularization were decreased in I/R,V,HPX and HZ groups(P<0.05).Compared with I/R group,the escape latency was significantly shortened at 2-7 days of reperfusion,the percentage o

关 键 词：血红素加氧酶-1 血液结合素 再灌注损伤 脑 

分 类 号：R743[医药卫生—神经病学与精神病学]