可卡因-苯丙胺调节转录肽对脑缺血-再灌注损伤小鼠脑保护机制的研究  

Effect of cocaine-amphetamine-regulated transcript peptide on cerebral protective mechanisms in cerebral ischemia/reperfusion injury in mice

作　　者：沙杜鹃[1] 韩勇[1] 顾双双[1] 王璐娜[1] 朱震寒[1] 李瑾[1] 李启明[1] 张均[1] 

机构地区：[1]南京大学医学院附属鼓楼医院急诊科,210008

出　　处：《中国脑血管病杂志》2012年第8期423-427,共5页Chinese Journal of Cerebrovascular Diseases

基　　金：南京市医学科技发展项目(YKK10069)

摘　　要：目的探讨可卡因-苯丙胺调节转录肽(CART)对缺血-再灌注小鼠脑梗死体积、脑组织8-羟基-2'-脱氧鸟苷(8-OHdG)及3-硝基酪氨酸(3-NT)水平的影响。方法选用10～12周龄的健康雄性昆明小鼠264只,随机分为缺血-再灌注组、CART组、等渗盐水组(各72只)及假手术组(48只)。建立大脑中动脉闭塞(MCAO)2 h,再灌注12、24、48及72 h模型(每时间点6只)。CART组于再灌注前经尾静脉注射CART55-102(0.5μg/kg,浓度12nmol/L;每24h重复给药1次),等渗盐水组给予同体积等渗盐水作对照。采用酶联免疫吸附实验双抗体夹心法测定脑组织中的8-OHdG和3-NT含量。结果①CART组再灌注后各时间点脑梗死体积均小于缺血-再灌注组和等渗盐水组,其中24、48和72 h差异均有统计学意义(P<0.01)。②与假手术组比较,缺血-再灌注组、CART组及等渗盐水组在再灌注后各时间点脑组织中8-OHdG和3-NT含量均升高。③与缺血-再灌注组、等渗盐水组比较,CART组再灌注后各时间点脑组织中的8-OHdG和3-NT含量均有所下调。其中8-OHdG仅在12 h差异有统计学意义(P<0.05),3-NT在24、48及72 h时间点差异均有统计学意义(P<0.05)。结论 CART对小鼠脑缺血-再灌注损伤的保护作用可能与下调8-OHdG及3-NT水平,抑制氧化应激有关。Objective To investigate the effect of cocaine-and amphetamine-regulated transcript peptide（CART） on the infarct volume and the levels of 8-hydroxy-2′-deoxyguanosine（8-OHdG） and 3-nitrotyrosine（3-NT） in cerebral ischemia/reperfusion injury in mice.Methods A total of 264 healthy male Kunming mice（10 to 12 weeks of age） were included in the study.They were randomly divided into four groups： ischemia/reperfusion,CART,isotonic saline（n=72 in each group）,and sham operation（n=48） groups.A model of middle cerebral artery occlusion（MCAO） for 2 hours and reperfusion for 12,24,48,and 72 hours was induced（n=6 for each time point）.Before reperfusion,CART55-102（0.5 μg,12 nmol/L;repeated every 24 hours） was injected via tail veins in the CART group,and the same volume of isotonic saline were given in the isotonic saline group for control.A double antibody sandwich enzyme-linked immunosorbent assay was used to detect the levels of 8-OHdG and 3-NT in brain tissue.Results ① The infarct volume at each time point in the CART group was lower than that in the isotonic saline and the ischemia/reperfusion groups,and there were significant differences at 24,48,and 72 hours（P0.01）.②Compared to the sham operation group,the levels of 8-OHdG and 3-NT increased at each time point after reperfusion in the ischemia/reperfusion,CART and isotonic saline groups.8-OHdG and 3-NT reached the peaks at 12 and 48 hours respectively.③Compared to the ischemia/reperfusion and isotonic saline groups,the levels of 8-OHdG and 3-NT in brain tissue were down-regulated at each time point after reperfusion in the CART group.There was only significant difference in 8-OHdG at 12 hours（P0.05） and there were significant difference in 3-NT at 24,48,and 72 hours（P0.05）.ConclusionThe protective effect of CART on cerebral ischemia/reperfusion injury in mice may be associated with the downregulation of 8-OHdG and 3-NT levels and the inhibition of oxidative stress.

关 键 词：脑梗死 再灌注损伤 小鼠 可卡因-苯丙胺调节转录肽 8-羟基-2'-脱氧鸟苷 3-硝基酪氨酸 

分 类 号：R743.3[医药卫生—神经病学与精神病学]