腺苷预处理对脑缺血再灌注损伤大鼠脑组织中微RNA-29b及凋亡蛋白B细胞淋巴瘤-2、磷酸化蛋白激酶B、人髓细胞白血病1表达的影响  

Effect of adenosine preconditioning on the expression of microRNA,B-lymphoma-2, phosphorylated protein kinase B and human myeloid leukemia-1 in brain tissue of rats with cerebral ischemia reperfusion injury

作　　者：曹武璟 季禾 谭军[2] 栗延伟[2] 秦全凯 CAO Wujing;JI He;TAN Jun;LI Yanwei;QIN Quankai(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;Department of Work Examination,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 [3]新乡医学院第三附属医院功能检查科,河南新乡453003

出　　处：《新乡医学院学报》2021年第12期1113-1120,共8页Journal of Xinxiang Medical University

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

摘　　要：目的观察腺苷预处理(AP)对大鼠脑缺血再灌注(IR)后脑组织中微RNA-29b(miR-29b)及凋亡蛋白B细胞淋巴瘤-2(Bcl-2)、磷酸化蛋白激酶B(p-Akt)、人髓细胞白血病1(Mcl-1)表达的影响,探讨AP对大鼠脑缺血再灌注损伤(CIRI)的保护作用及机制。方法将健康雄性清洁级Sprague Dawley大鼠135只随机分为假手术组、IR组、AP组,每组45只。AP组大鼠在手术前3 d腹腔注射2 mL腺苷注射液(1.5 mg·kg^(-1)),每日1次;假手术组和IR组大鼠在手术前3 d每日相同时间点腹腔注射2 mL生理盐水。IR组及AP组大鼠采用改良线栓法行脑中动脉栓塞并于栓塞2 h后拨出栓线恢复血供制备IR模型,假手术组大鼠不做颈动脉结扎,其余手术方法同IR组和AP组。根据5分制神经行为学评分标准对大鼠进行神经功能损伤评分;术后24 h分别取各组造模成功大鼠,断头处死,应用氯化三苯四氮唑染色法观察脑梗死情况,并计算脑梗死体积占比;苏木精-伊红染色观察大鼠海马区神经元细胞存活情况,实时荧光定量聚合酶链反应法检测各组大鼠海马组织内miR-29b表达,Western blot法检测大鼠海马组织中Bcl-2、p-Akt、Mcl-1表达。结果假手术组、IR组和AP组大鼠神经行为学评分和脑梗死体积占比比较差异有统计学意义(F=186.500、191.300,P<0.05),IR组和AP组大鼠神经行为学评分和脑梗死体积占比显著高于假手术组(P<0.05),AP组大鼠神经行为学评分和脑梗死体积占比显著低于IR组(P<0.05)。假手术组大鼠脑皮质神经元形态结构正常;IR组大鼠脑皮质神经元排列紊乱、数目明显减少、结构疏松,细胞体积变小,细胞核溶解、出现空泡,细胞膜完整性破坏,细胞呈三角形或梭形,间质水肿;与IR组比较,AP组大鼠神经元损伤显著减轻。手术后2、6、24、48 h, IR组和AP组大鼠海马组织miR-29b相对表达量显著高于假手术组(P<0.05),AP组大鼠海马组织miR-29b相对表达量显著高于IR组(P<0.05)。�Objective To observe the adenosine preconditioning(AP) on the expressions of microRNA-29 b(miR-29 b),B-lymphoma-2(Bcl-2),phosphorylated protein kinase B(p-Akt) and human myeloid leukemia-1(Mcl-1) in brain tissue of rats after cerebral ischemia reperfusion(IR),and to investigate the protective effect and mechanism of AP on cerebral ischemia reperfusion injury(CIRI) in rats.Methods One hundred and thirty-five healthy male Sprague Dawley rats were randomly divided into sham operation group, IR group and AP group, with 45 rats in each group.The rats in the AP group were intraperitoneally injected with 2 mL adenosine injection(1.5 mg· kg^(-1)) once a day for 3 days before operation, while the rats in the sham operation group and IR group were intraperitoneally injected with 2 mL saline at the same time point for 3 days before operation.The rats in the IR group and AP group were underwent middle cerebral artery occlusion with improved thread embolization method, and the thread was pulled out at 2 hours after embolization to prepare the IR models.The rats in the sham operation group were not ligated, and other operation methods were same as those in the IR group and AP group.The neurobehavioral scoring standard of 5-point system was used to evaluate the neurological injury of rats;24 hours after operation, the rats in each group were decapitated and killed, and the brain tissue was taken, the cerebral infarction was observed by triphenyltetrazolium chloride staining, and the proportion of cerebral infarction volume was calculated.The survival of hippocampal neurons was observed by hematoxylin-eosin staining, the expression of miR-29 b in brain tissue was detected by real-time fluorescence quantitative polymerase chain reaction, and the expression of Bcl-2,p-Akt and Mcl-1 protein in hippocampus were detected by Western blot.Results There were significant differences in the neurobehavioral score and the proportion of cerebral infarction volume of rats among the sham group, IR group and AP group(F=186.500,191.300;P<0.05).

关 键 词：脑缺血再灌注损伤 腺苷预处理 细胞凋亡 微RNA-29b 凋亡蛋白 

分 类 号：R743[医药卫生—神经病学与精神病学]