腺苷预处理抑制内质网应激对大鼠脑缺血再灌注损伤的保护作用  

Adenosine preconditioning inhibits endoplasmic reticulum stress and protects against cerebral ischemia-reperfusion injury in rats

作　　者：薛倩倩 李国栋 栗延伟[1] 谭军[1] XUE Qianqian;LI Guodong;LI Yanwei;TAN Jun(不详;Department of Neurology,the Third Affiliated Hospital of Xinxiang Medical University,Xinxiang Henan 453000,China)

机构地区：[1]新乡医学院第三附属医院神经内科,453000

出　　处：《中国神经免疫学和神经病学杂志》2024年第5期352-357,共6页Chinese Journal of Neuroimmunology and Neurology

基　　金：河南省医学科技攻关计划联合共建项目(编号:LHGJ20230546)。

摘　　要：目的研究腺苷预处理对大鼠脑缺血再灌注损伤(cerebral ischemia reperfusion injury,CIRI)后内质网应激(endoplasmic reticulum stress,ERS)标志蛋白葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)及肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-1β、核因子κB(NF-κB)表达的影响,进而探讨腺苷对CIRI的神经保护作用。方法选取健康雄性SD大鼠36只,体重250~280 g,按照随机数字法将大鼠分成假手术组、缺血再灌注(I/R)模型组和腺苷预处理(adenosine pretreatment)组,每组各12只。采用线栓法构建大鼠大脑中动脉栓塞模型(middle cerebral artery occlusion,MCAO),腺苷预处理组于制模前3 d按体重1.5 mg/kg腹腔注射腺苷注射液2 mL,1次/d,连续3 d。栓塞2 h后对大鼠进行神经功能缺损评分(neurological severity score,NSS)。再灌注24 h后处死大鼠,断头取脑,采用TTC染色法观察各组大鼠脑梗死体积,采用RT-qPCR法检测各组大鼠脑组织GRP78和CHOP mRNA表达,采用ELISA试剂盒检测大鼠脑组织TNF-α、IL-6、IL-1β表达水平,采用Western blot法检测大鼠脑组织NF-κB蛋白表达水平。结果I/R模型组大鼠NSS,脑组织GRP78 mRNA、CHOP mRNA、TNF-α、IL-6、IL-1β、IκB蛋白、p65蛋白表达水平较假手术组高(P<0.01或P<0.05);腺苷预处理组大鼠脑梗死体积、NSS较I/R模型组低(P<0.05),脑组织GRP78 mRNA、CHOP mRNA、TNF-α、IL-6、IL-1β、IκB蛋白、p65蛋白表达水平较I/R模型组低(P<0.01或P<0.05)。结论腺苷预处理可减轻I/R导致的神经细胞继发性损伤,其作用机制可能与腺苷抑制ERS有关。Objective To study the effect of pretreatment with adenosine on the expression of endoplasmic reticulum stress(ERS)glucose-regulated protein 78(GRP78),C/EBP homologous protein(CHOP)and tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1βand nuclear factor kappa-B(NF-κB)after cerebral ischemia reperfusion injury(CIRI)in rats,in order to investigate the neuroprotective effect of adenosine on CIRI.Methods Thirty-six healthy male Srague Dawley(SD)rats weighing 250-280 g were randomly divided into three groups and 12 rats in each group:sham group,ischemia reperfusion(I/R)group,and adenosine preconditioning(AP)group.The rat model of middle cerebral artery occlusion(MCAO)was established by thread embolization.In AP group,2 mL of adenosine was injected intraperitoneally at 1.5 mg/kg body weight 3 days before making model,once a day for 3 days.The neurological severity score(NSS)was measured after 2 h of embolization.After 24 h of reperfusion,the rats were killed and the brains were harvested.TTC stent method was used to observe the infarct volume of the rat's brain.The mRNA expression of GRP78 and CHOP was detected by RT-qPCR.The expression of TNF-α,IL-6 and IL-1βin brain tissue were measured by ELISA and Western blot was used to measure the expression of NF-κB protein.Results The expression levels of NSS,GRP78 mRNA,CHOP mRNA,TNF-α,IL-6,IL-1β,IκB protein and p65 protein in brain tissue of rats in I/R model group were higher than those in sham group(all P<0.05);The infarct volume and NSS in AP group were lower than those in I/R model group(both P<0.05).The expression levels of GRP78 mRNA,CHOP mRNA,TNF-α,IL-6,IL-1β,IκB protein and p65 protein in brain tissue in AP group were lower than those of I/R model group(P<0.05).Conclusions AP can alleviate the secondary injury of nerve cells induced by I/R,and its mechanism may be related to the inhibition of ERS by adenosine.

关 键 词：脑梗死 脑缺血 再灌注损伤 腺苷 内质网应激 炎症 核因子ΚB 

分 类 号：R743.33[医药卫生—神经病学与精神病学]