腺苷预处理对大鼠脑缺血再灌注损伤的保护作用  

Protective effect of adenosine preconditioning on cerebral ischemia-reperfusion injury in rats

作　　者：庞艳利 尉娜[2] 谭军[2] PANG Yanli;WEI Na;TAN Jun(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》2020年第2期125-130,共6页Journal of Xinxiang Medical University

基　　金：新乡市科技局科技创新人才计划项目(编号:CXRC17003)。

摘　　要：目的探讨腺苷预处理对大鼠脑缺血再灌注损伤的保护作用。方法 36只健康雄性Sprague Dawley大鼠随机分为假手术组、缺血再灌注组和腺苷预处理组,每组12只。术前3 d,腺苷预处理组大鼠按照1.5 mg·kg^-1将腺苷用生理盐水稀释至2 mL给予腹腔注射,假手术组和缺血再灌注组大鼠腹腔注射生理盐水2 mL,均每日1次。缺血再灌注组、腺苷预处理组大鼠应用栓线法制备大鼠大脑中动脉栓塞模型;假手术组不阻塞大脑中动脉,余步骤同缺血再灌注组及腺苷预处理组。采用5分制评分法对各组大鼠进行神经功能缺损评分。每组随机选取6只大鼠麻醉处死取脑组织,2,3,5-氯化三苯基四氮唑染色观察大鼠的脑梗死体积占比。每组另取6只大鼠进行心脏灌注,取脑组织制备切片,苏木精-伊红染色法观察脑组织的病理改变,免疫组织化学染色法检测脑组织中Caspase-3蛋白阳性细胞表达率,末端脱氧核苷酸转移酶介导的脱氧尿嘧啶核苷三磷酸缺口末端标记法检测脑组织中细胞凋亡率。结果假手术组大鼠脑组织神经元排列整齐,细胞形态及细胞核完好,间质未见明显水肿;缺血再灌注组大鼠可见神经元排列紊乱、变性及坏死,细胞核固缩、深染,间质明显水肿;腺苷预处理组大鼠脑组织神经元损伤程度较缺血再灌注组减轻。3组大鼠神经功能缺损评分、脑梗死体积比、脑组织中Caspase-3蛋白阳性细胞表达率、细胞凋亡率比较差异有统计学意义(F=98. 036、298. 544、33. 426、90. 165,P <0. 05);其中缺血再灌注组、腺苷预处理组大鼠神经功能缺损评分、脑梗死体积比、Caspase-3蛋白阳性细胞表达率、细胞凋亡率显著高于假手术组(P <0. 05),腺苷预处理组大鼠神经功能缺损评分、脑梗死体积比、Caspase-3蛋白阳性细胞表达率、细胞凋亡率显著低于缺血再灌注组(P <0. 05)。结论腺苷预处理可能通过抑制Caspase-3蛋白表达�Objective To investigate the protective effect of adenosine pretreatment on cerebral ischemia-reperfusion injury of rats. Methods Thirty-six healthy male Sprague Dawley rats were randomly divided into sham operation group,ischemia-reperfusion group and adenosine pretreatment group,with 12 rats in each group. the rats in the adenosine pretreatment group were given adenosine(1. 5 mg · kg^-1)(adenosine was diluted to 2 mL with normal saline) by intraperitoneal injection before operation;the rats in the sham operation group and ischemia-reperfusion group were intraperitoneally injected with 2 mL of normal saline;once a day for 3 days. The rats in the ischemia-reperfusion group and adenosine pretreatment group were made the middle cerebral artery occlusion model by the thread plug method;the rats in the sham operation group did not block the middle cerebral artery,and the other steps were the same as those in the ischemia-reperfusion group and adenosine pretreatment group. The neurological deficits of rats in each group was evaluated by five-point scoring method. Six rats in each group were randomly selected for anesthesia and then the rats were sacrificed to take the brain sections. The proportion of cerebral infarction volume was observed by 2,3,5-triphenyltetrazolium chloride staining. Another 6 rats in each group were performed cardiac perfusion,and the brains tissues were taken to prepare sections. The pathological changes of brain tissue were observed by hematoxylin-eosin staining;the positive expression rate of Caspase-3 protein in brain tissue was detected by immunohistochemical staining;and the brain cell apoptosis rate was detected by Td T-mediated d UTP nick end labeling method. Results In the sham operation group,the neurons of the brain tissue were arranged neatly,the cell morphology and nucleus were intact,and no obvious edema was found in the stroma. In the ischemia-reperfusion group,neurons were arranged disorderly,degeneration and necrosis,the nucleus contracted and deep stained,and the interstitial

关 键 词：大鼠 腺苷 脑 缺血再灌注损伤 CASPASE-3 细胞凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]