HIF-2α通过抑制炎症反应减轻大鼠脑出血损伤  

HIF-2α attenuates intracerebral hemorrhage injury in rats by inhibiting inflammatory response

作　　者：陈辉 甘荟 肖涵 张莉[1] 王璐[1] 蒋宁 翟瑄[1] Chen Hui;Gan Hui;Xiao Han;Zhang Li;Wang Lu;Jiang Ning;Zhai Xuan(Department of Neurosurgery,Chongq ing Key Laboratory of Pediatrics,Minis try of Education Key Laboratory of Child Development and Disorders,National Clinical Research Center for Child Health and Disorders(Chongqing),China International Science and Technology Cooperation base of Child development and Critical Disorders,Children's Hospital of Chongqing Medical University;Department of Pathology and Pathophysiology,College of Basic Medicine,Chongqing Medical University)

机构地区：[1]重庆医科大学附属儿童医院神经外科,儿科学重庆市重点实验室,儿童发育疾病研究教育部重点实验室,国家儿童健康与疾病临床医学研究中心(重庆),儿童发育重大疾病国家国际科技合作基地,重庆400014 [2]重庆医科大学基础医学院病理与病理生理学教研室,重庆400016

出　　处：《重庆医科大学学报》2020年第1期65-70,共6页Journal of Chongqing Medical University

基　　金：重庆市科委基础科学与前沿技术研究重点资助项目(编号:cstc2015jcyjBX0144);中国抗癫痫协会癫痫科研基金——UCB基金资助项目(编号:2018003)。

摘　　要：目的:探讨缺氧诱导因子2α(hypoxia-inducible factor 2α,HIF-2α)在脑出血损伤后(intracerebral hemorrhage,ICH)的作用,并明确其是否参与调控脑出血后的炎症反应。方法:健康雄性SD大鼠95只,其中35只随机分配用于以下时间点分析:Sham、12 h、24 h、48 h、3 d、5 d和7 d,每组5只;另外60只随机分为Sham组(假手术组,注射等量生理盐水)、ICH组(脑出血模型组组,胶原酶诱导的脑出血模型)、Vehicle组(空载体组组,建模前注射空载慢病毒载体)和Oe-HIF-2α组(HIF-2α过表达组,建模前注射HIF-2α过表达慢病毒载体)用于脑含水量检测、神经功能评分、免疫印记分析和免疫荧光检测。通过脑含水量和Garcia神经功能评分评估脑损伤的严重程度;免疫印迹检测HIF-2α及肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)、白细胞介素-18(interleukin 18,IL-18)和白细胞介素-1β(interleukin 1β,IL-1β)的表达水平,免疫荧光检测病灶周边髓过氧化物酶(myeloperoxidase,MPO)的表达。结果:HIF-2α表达在ICH后24 h(0.555 4±0.070 2,P=0.000)开始上调并于3 d(2.368 4±0.346 6,P=0.000)达到峰值,随后降低;与ICH组相比较,Oe-HIF-2α组脑3 d时含水量明显降低(0.793 5±0.002 5,P=0.000),神经功能明显改善(14.700 0±0.674 9,P=0.000);与ICH组相比,Oe-HIF-2α组TNF-α(1.350 4±0.191 5,P=0.000)、IL-1β(1.158 4±0.070 8,P=0.000)、IL-18(0.784 2±0.073 9,P=0.000)等炎性介质在3 d时表达明显降低;Oe-HIF-2α组(3.500 0±0.534 5)病灶周边区域MPO表达明显低于ICH组(5.125 0±0.991 0,P=0.002)。结论:脑出血后,HIF-2α表达上调并且通过抑制TNF-α、IL-1β、IL-18等炎症因子的表达,抑制炎症反应减轻大鼠脑出血损伤。Objective:To investigate the role of hypoxia-inducible factor 2α(HIF-2α)in intracerebral hemorrhage(ICH)injury,and to determine whether HIF-2α is involved in the regulation of inflammatory response after ICH. Methods:There were 95 healthy male Sprague-Dawley rats,and 35 of them were randomly assigned to seven groups for different data collection time points:Sham,12 h,24 h, 48 h,3 d,5 d,and 7 d,with 5 rats in each group. The other 60 rats were randomly divided into Sham group(sham operation group with injection of the same amount of normal saline),ICH group(ICH model group with collagenase-induced ICH),Vehicle group(empty vector group with injection of empty lentiviral vector before modeling),and Oe-HIF-2α group(HIF-2α overexpression group with injection of HIF-2α overexpression lentiviral vector before modeling)for brain water content mea-surement,neurological function scoring,Western blot analysis,and immunofluorescence detection. The severity of brain injury was assessed by brain water content and Garcia neurological deficit scores. The expression levels of HIF-2α,tumor necrosis factor α(TNF-α),interleukin 18(IL-18),and interleukin 1 beta(IL-1β)were determined by Western blot,and the expression of myeloperoxidase(MPO)was determined by immunofluorescence assay. Results:The expression of HIF-2α began to increase at 24 hours after ICH(0.555 4±0.070 2,P=0.000),peaked on day 3(2.368 4±0.346 6,P=0.000),and then decreased. Compared with the ICH group on day 3,the Oe-HIF-2α group had significantly decreased brain water content(0.793 5±0.002 5,P=0.000),significantly improved neurological function(14.700 0±0.674 9,P=0.000),and significantly reduced expression of inflammatory mediators including TNF-α(1.350 4±0.191 5,P=0.000),IL-1β(1.158 4±0.070 8,P=0.000),and IL-18(0.784 2±0.073 9,P=0.000);the expression of MPO in the surrounding area of lesion was significantly lower in the Oe-HIF-2α group than in the ICH group(3.500 0±0.534 5 vs. 5.125 0±0.991 0,P=0.002). Conclusion:After ICH,HIF-2α is upregula

关 键 词：缺氧诱导因子2α 炎症 神经保护 脑出血 

分 类 号：R741.02[医药卫生—神经病学与精神病学]