腺苷预处理对脑缺血再灌注大鼠小胶质细胞极化及神经损伤的影响  

Effect of adenosine pretreatment on microglia polarization and nerve injury in rats with cerebral ischemia reperfusion

作　　者：费增焱 栗延伟[2] 谭军[2] 薛倩倩 FEI Zengyan;LI Yanwei;TAN Jun;XUE Qianqian(The Third Clinical College of Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Department of Neurology,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang 453003,Henan Province,China)

机构地区：[1]新乡医学院第三临床学院,河南新乡453003 [2]新乡医学院第三附属医院神经内科,河南新乡453003

出　　处：《新乡医学院学报》2024年第6期501-507,共7页Journal of Xinxiang Medical University

基　　金：河南省医学科技攻关计划联合共建项目(编号:LHGJ20200533)。

摘　　要：目的探讨大鼠脑缺血再灌注(IR)损伤后小胶质细胞表型的变化以及腺苷对脑IR损伤大鼠的神经损伤的作用。方法将36只健康雄性Sprague Dawley大鼠按随机数字表法分为假手术(Sham)组、IR组和腺苷预处理(AP)组,每组12只。AP组大鼠造模前每天腹腔注射2 mL腺苷注射液,连续3 d;Sham组及IR组大鼠每天腹腔注射2 mL生理盐水,连续3 d。IR组和AP组大鼠采用线栓法构建大脑中动脉栓塞模型;Sham组大鼠仅分离颈动脉,不结扎血管。造模2 h后,采用5分制神经行为学评分对各组大鼠进行神经行为学评估。恢复大脑中动脉血流灌注24 h后处死各组大鼠,取出脑组织,采用苏木精-伊红(HE)染色观察缺血半暗带区域脑组织形态学改变,免疫荧光染色法检测M1型小胶质细胞标志物和M2型小胶质细胞标志物的共表达情况,采用实时荧光定量聚合酶链式反应法(qRT-PCR)检测2组大鼠脑组织中M1型小胶质细胞释放的促炎因子诱导型一氧化氮合酶(iNOS)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β和M2型小胶质细胞释放的抗炎因子IL-4、IL-10、转化生长因子-β(TGF-β)的相对表达量。结果IR组及AP组大鼠神经行为学评分显著高于Sham组,IR组大鼠神经行为学评分显著高于AP组(P<0.05)。HE染色结果显示,Sham组大鼠脑组织中细胞结构完整,细胞核清晰可见,细胞排列紧密,无间质水肿;IR组大鼠脑组织中细胞结构明显破坏,细胞排列不整齐,间质疏松,胞体内出现空泡;AP组大鼠脑组织中细胞结构较IR组完整,正常细胞数量较多,排列较整齐,细胞核较完整。免疫荧光染色结果显示,IR组、AP组大鼠脑缺血半暗带区域M1型、M2型小胶质细胞数量显著高于Sham组,IR组大鼠M1型小胶质细胞数量显著高于AP组,M2型小胶质细胞数显著低于AP组(P<0.05)。qRT-PCR结果显示,IR组、AP组大鼠脑组织中促炎细胞因子TNF-α、IL-1β、iNOS和抗炎细胞因子IL-4、IL-10、TGF-β�Objective To investigate the changes in microglia phenotype after cerebral ischemia reperfusion(IR)injury and the effects of adenosine on nerve injury of cerebral IR injured rats.Methods Thirty-six healthy male Sprague Dawley rats were randomly divided into the sham operation(Sham)group,IR group,and adenosine pretreatment(AP)group,with 12 rats in each group.Before modeling,rats in the AP group were intraperitoneally injected with 2 mL of adenosine injection daily for 3 consecutive days,and rats in the Sham group and IR group were intraperitoneally injected with 2 mL of normal saline daily for 3 consecutive days.The middle cerebral artery occlusion models of rats in the IR group and AP group were constructed by using the suture-occluded method,and only the carotid artery of rats was isolated in the Sham group without ligation of blood vessels.At 2 hours after modeling,the neuroethology of rats in each group were evaluated according to a 5-point neurobehavioral scale.At 24 hours after restoring the blood perfusion in the middle cerebral artery,the rats in each group were executed,and their brain tissues were removed.The morphological changes of the brain tissues in the ischemic penumbra region were observed after hematoxylin-eosin(HE)staining.The co-expression of M1-type microglia markers and M2-type microglia markers was detected by immunofluorescence staining.The relative expression levels of pro-inflammatory factors inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α)and interleukin(IL)-1βreleased by M1-type microglia,and anti-inflammatory factors IL-4,IL-10 and transforming growth factor-β(TGF-β)released by M2-type microglia were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Results The neurobehavioral scores of rats in the IR group and AP group were significantly higher than those in the Sham group,and the neurobehavioral score of rats in the IR group was significantly higher than that in the AP group(P<0.05).HE staining results showed that the brain cells of rats i

关 键 词：腺苷预处理 脑缺血再灌注 M1型小胶质细胞 M2型小胶质细胞 炎症 

分 类 号：R743.3[医药卫生—神经病学与精神病学]