外源性硫化氢通过抑制闭合蛋白和闭锁小带蛋白1表达减轻脑缺血再灌注大鼠脑水肿和脑损伤  

Exogenous hydrogen sulfide attenuates brain edema and injury of focal cerebral ischemia- reperfusion in rats by inhibiting the expressions of occludin and zonula occluder^-I protein

作　　者：魏霞[1] 潘红[1] 姚旋[1] 张兵[1] 

机构地区：[1]哈尔滨医科大学附属第二医院麻醉科,150086

出　　处：《国际脑血管病杂志》2015年第12期893-898,共6页International Journal of Cerebrovascular Diseases

基　　金：黑龙江省教育厅科学技术研究项目（12541423）

摘　　要：目的探讨外源性硫化氢对脑缺血再灌注后脑水肿和脑损伤的影响及其机制。方法60只雄性SD大鼠随机分为假手术组、缺血再灌注组、硫化氢30ppm组（1ppm=1mg／L）和硫化氢60ppm组，每组15只。采用大脑中动脉闭塞法建立大鼠局部脑缺血2h再灌注24h模型。在脑缺血再灌注24h后观察神经功能学评分，测定脑梗死体积、脑水肿程度以及血脑屏障通透性变化。采用蛋白质印迹法检测缺血半暗带闭合蛋白和闭锁小带蛋白1（zonula occludens protein1，ZO-1）表达。结果与缺血再灌注组比较，吸入硫化氢30ppm和60ppm能以剂量依赖方式显著增高神经功能评分（P均〈0．05），缩小脑梗死体积（P均〈0．05），减轻脑水肿（P均〈0．05）。缺血再灌注组缺血侧脑组织伊文思蓝含量较假手术组显著增高[（0．74±0．14）μg／100mg对（0．19±0．06）μg／100mg；P〈0．05]，硫化氢30ppm组[（0．554-0．10）μg／100mg]和硫化氢60ppm组[（0．35±0．08）μg／100mg]脑组织伊文思蓝含量均较缺血再灌注组显著降低（P均〈0．05），其中硫化氢60ppm组显著低于硫化氢30ppm组（P〈0．05）。蛋白质印迹分析显示，硫化氢30ppm和60ppm组半暗带闭合蛋白（Q621％±0．101％对0．787％±0．087％对0．453％±0．127％；P〈0．05）和ZO-1（0．602％±0．118％对0．778％±0．805％对0．426％±0.146％；P〈0．05）的表达水平均较缺血再灌注组显著增加，其中硫化氢60ppm组闭合蛋白和ZO-1的表达水平均显著高于硫化氢30ppm组（P均〈0．05）。结论吸入外源性硫化氢能以剂量依赖方式显著降低脑缺血再灌注后脑水肿，缩小脑梗死体积并改善神经功能，其机制可能与抑制闭合蛋白和ZO-1表达、下调维持血脑屏障完整性有关。Objective To investigate the effects of exogenous hydrogen sulfide on bralu edema and injury and their mechanisms. Methods Sixty male SD rats were randomly divided into a sham operation group, an ischemia-reperfusion group, a 30 ppm hydrogen sulfide group, and a 60 ppm hydrogen sulfide group （n = 15 in each group; 1 ppm = 1 mg/L）. A model of focal cerebral ischemia for 2 h and reperfusion for 24 h was induced by middle cerebral artery occlusion. The neurological scores were observed after 24 h cerebral ischemia- reperfusion. The cerebral infarction volume, the degree of brain edema, and the changes of blood-brain barrier permeability were measured. Western blotting was used to detect the expressions of occludin and zonula occludens-1 protein （ZO-1） in ischemic penumbra. Results Compared with the ischemia-reperfusion group, the neurological function scores in the 30 ppm and 60 ppm hydrogen sulfide group significantly increased in a dose- dependent manner （all P 〈0. 05） and the neurological deficits and infarct voktrne reduced （all P 〈0. 05）, and the brain edema alleviated （all P 〈 0. 05）. The content of Evans blue in the ischemic brain tissue in the ischemia- reperfusion group increased significantly compare with the sham operation group （0. 74 ± 0. 14 μg/100 mg vs. 0. 19 ±0. 06 μg/100 mg; P 〈0. 05）. The content of Evans blue in the brain tissue in the 30 ppm hydrogen sulfide group （0. 55 ± 0. 10 μg/100 mg） and the 60 ppm hydrogen sulfide group （0. 35 ± 0. 08 μg/100 mg） decreased significantly compared with the ischemia-reperfusion group （all P 〈0. 05）, among them the 60 ppm hydrogen sulfide group was significantly lower than the 30 ppm hydrogen sulfide group （P〈0. 05）. Western blot analysis showed that expression levels of occludin in penumbra （0. 621%±0. 101% vs. O. 787%±0. 087% vs. O. 453% f 0. 127%; P〈0.05） and ZO-1 （0.602%±0. 118% vs. 0.778%±0.805% vs. 0.426% ±0. 146; P〈0.05） in the 30 ppm and 60 ppm hydrogen sulfide groups increased

关 键 词：硫化氢 脑缺血 再灌注损伤 脑水肿 血脑屏障 闭合蛋白 闭锁小带蛋白1 神经保护药 大鼠 

分 类 号：R743[医药卫生—神经病学与精神病学]