HIF-1α/BNIP3信号通路在IL-4减轻小鼠脑缺血再灌注损伤中的作用及其与自噬的关系  

Role of HIF-1α/BNIP 3 signaling pathway in IL-4 attenuating cerebral ischemia-reperfusion injury in mice and its relationship with autophagy

作　　者：雷爱弟 张健莉 周衡 LEI Aidi;ZHANG Jianli;ZHOU Heng(Department of Neurology,Xiamen Fifth Hospital,Xiamen 361101,China)

机构地区：[1]厦门市第五医院神经内科,厦门361101 [2]北京天坛医院神经病学中心神经感染与免疫科,北京100071

出　　处：《中国免疫学杂志》2023年第10期2149-2153,2159,共6页Chinese Journal of Immunology

摘　　要：目的:探究低氧诱导因子-1α/Bcl-2/腺病毒E1B-19k Da相互作用蛋白3(HIF-1α/BNIP3)在IL-4减轻小鼠脑缺血再灌注损伤(CIRI)中的作用及其与自噬的关系。方法:将SPF级BALB/c小鼠采用随机数字表法分为假手术组、缺血再灌注组(CIRI组)、IL-4组、HIF-1α抑制剂2ME2组(2ME2组)和IL-4+HIF-1α抑制剂2ME2组(IL-4+2ME2组)。采用线栓法构建CIRI组、IL-4组、2ME2组和IL-4+2ME2组大脑中动脉栓塞(MACO)模型,闭塞缺血60 min后再灌注24 h,假手术组除不阻断小鼠大脑中动脉外,其余操作同上。IL-4组、2ME2组、IL-4+2ME2组建模前30 min分别腹腔注射对应的IL-4C复合体溶液及尾静脉注射2ME2溶液。再灌注24 h后进行神经行为学评分,然后处死小鼠取脑组织。采用TTC染色检测脑梗死体积;干湿重法测定脑组织含水量;TUNEL染色检测细胞凋亡;透射电镜计数自噬小体;比色法测定SOD、MDA和ROS水平;Western blot检测HIF-1α、BNIP3、LC3-Ⅱ和Beclin-1表达。结果:与假手术组相比,CIRI组小鼠神经行为学评分、脑梗死体积、脑含水量、细胞凋亡率、自噬小体数量、MDA、ROS、HIF-1α、BNIP3、LC3-Ⅱ和Beclin-1水平升高,SOD水平降低(P<0.05);与CIRI组相比,IL-4组神经行为学评分、脑梗死体积、脑含水量、细胞凋亡率、MDA、ROS水平降低,自噬小体数量、SOD、HIF-1α、BNIP3、LC3-Ⅱ和Beclin-1水平升高(P<0.05);与IL-4组相比,2ME2组、IL-4+2ME2组神经行为学评分、脑梗死体积、脑含水量、细胞凋亡率、自噬小体数量、MDA、ROS、HIF-1α、BNIP3、LC3-Ⅱ和Beclin-1水平降低,SOD水平升高(P<0.05)。结论:IL-4可通过激活HIF-1α/BNIP3信号通路,激活细胞自噬,减少细胞凋亡和氧化应激,从而减轻小鼠CIRI。Objective:To explore the role of hypoxia-inducible factor-1α/Bcl-2/adenovirus E1B-19kDa interacting protein 3(HIF-1α/BNIP3)in IL-4-induced attenuation of cerebral ischemia-reperfusion injury(CIRI)in mice and its relationship with autophagy.Methods:SPF BALB/c mice were divided into sham group,cerebral ischemia reperfusion injury group(CIRI group),IL-4 group,HIF-1αinhibitor 2ME2 group(2ME2 group)and IL-4+HIF-1αinhibitor 2ME2 group(IL-4+2ME2 group)by random number table method.Middle cerebral artery embolism(MACO)models of CIRI group,IL-4 group,2ME2 group and IL-4+2ME2 group were constructed by thread embolization method.Occlusion of ischemia was performed for 60 minutes and then reperfusion for 24 hours.In sham-operated group,except that the middle cerebral artery of mice was not blocked,the other operations were the same as above.IL-4 group,2ME2 group and IL-4+2ME2 group were intraperitoneally injected with the corresponding IL-4C complex solution and the tail vein injection with 2ME2 solution 30 minutes before modeling.After 24 hours of reperfusion,neurobehavioral scores were performed,then the mice was sacrificed and the brain tissues were taken.TTC staining was used to detect cerebral infarction volume;dry and wet weight method was used to determine brain tissue water content;TUNEL staining was used to detect cell apoptosis;transmission electron microscopy was used to count autophagosomes;colorimetric method was used to detect SOD,MDA and ROS levels;Western blot was used to detect expressions of HIF-1α,BNIP3,LC3-Ⅱand Beclin-1.Results:Compared with sham group,neurobehavioral score,cerebral infarction volume,brain water content,apoptosis rate,number of autophagosomes,and levels of MDA,ROS,HIF-1α,BNIP3,LC3-Ⅱ,Beclin-1 in CIRI group were increased,while level of SOD was decreased(P<0.05);compared with CIRI group,neurobehavioral score,cerebral infarction volume,brain water content,cell apoptosis rate,MDA and ROS levels in IL-4 group were decreased,the number of autophagosomes,SOD,HIF-1α,BNIP3,LC3-Ⅱand Be

关 键 词：低氧诱导因子-1α/Bcl-2/腺病毒E1B-19kDa相互作用蛋白3 白介素-4 脑缺血再灌注损伤 自噬 

分 类 号：R392.2[医药卫生—免疫学]