胡须刺激通过调节神经可塑性改善小鼠桶状皮质缺血性脑卒中后的神经功能缺损  

Protective Effect of Whisker Stimulation on Focal Barrel Cortex Ischemic Stroke in Mice

作　　者：孙金梅[1] 张成杰[1] 李尧[1] 陈彬[1] SUN Jin-mei;ZHANG Cheng-jie;LI Yao;CHEN Bin(Department of Neurology,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院神经内科,北京100050

出　　处：《神经损伤与功能重建》2022年第6期311-314,336,共5页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金(No.81801334)。

摘　　要：目的:探讨规律胡须刺激是否可改善小鼠桶状皮质(Barrel Cortex)局灶性脑缺血后神经功能并从神经可塑性角度探讨其机制。方法:60只雄性C57BL/6随机分成假手术组、脑梗死组和胡须刺激组,每组20只。胡须刺激组在小鼠桶状皮质局灶性脑缺血模型建立后第3天开始给予规律胡须刺激(15 min/次,3次/d,共12 d)。3组小鼠均在造模后3、7、14 d进行贴纸去除试验;造模后14 d进行HomeCage行为学检测、采用Western blot技术检测脑梗死灶周围突触素(SPY)-1、生长相关蛋白43(GAP 43)、脑源性神经营养因子(BDNF)和神经丝(NF)蛋白表达水平,免疫荧光染色检测NF纤维。结果:脑梗死造模后3、7、14 d,脑梗死组小鼠的贴纸感知时间和贴纸去除时间均高于假手术组(均P<0.05);造模后7 d,胡须刺激组小鼠的贴纸去除时间短于脑梗死组(P<0.05);造模后14 d,胡须刺激组小鼠的贴纸感知时间和贴纸去除时间均短于脑梗死组(均P<0.05)。HomeCage行为学分析结果显示,与假手术组相比,脑梗死组和胡须刺激组小鼠活跃性指标均有明显下降,而不动时间明显延长(均P<0.05);与脑梗死组相比,胡须刺激组小鼠活跃性指标均明显提高,不动时间明显缩短(均P<0.05)。Western blot结果显示,造模后14 d,小鼠脑梗死灶周围SYP-1和GAP-43蛋白的表达明显上调,给予胡须刺激后上调更加明显(均P<0.05);梗死灶周围NF的表达明显下降,给予规律的胡须刺激可逆转该改变(P<0.05)。免疫组化化学染色结果提示梗死灶周围NF形态紊乱,而胡须刺激后NF排列紊乱现象明显改善,染色强度明显增加(P<0.05)。结论:规律的胡须刺激有助于改善小鼠桶状皮质局灶性脑缺血后神经功能缺损症状,推测部分机制与梗死灶周围SYP-1、GAP-43和NF等参与的神经可塑性有关。Objective:To investigate the protective effect of whisker stimulation on barrel cortex ischemic stroke in mice and explore its mechanism.Methods:A total of 60 male C57BL/6 mice were randomly divided into the sham-operation group,ischemia group,and treatment group with 20 mice in each.The treatment group underwent establishment of the focal barrel cortex ischemia model,then 3 days later,received regular whisker stimulation(15 min/time,3 times/day) for 12 days.At 3,7,and 14 days after model establishment,the sticker removal test was performed in all 3 groups.At 14 days after model establishment,the HomeCage behavior test was performed;western blot was used to assess the expression levels of synaptophysin 1(SYP-1),growth-associated protein 43(GAP-43),brain-derived neurotrophic factor(BDNF),and neurofilament(NF) in the penumbral tissue;and immunofluorescent staining was applied to observe NF structure.Results:At 3,7,and 14 days after ischemia model establishment,the ischemia group mice showed prolonged sticker contact time and removal time compared to the sham-operation group mice(all P<0.05).At 7 days after model establishment,the treatment group presented a shorter sticker removal time than the ischemia group(P<0.05);14 days after model establishment,the treatment group showed both a shorter sticker contact time and shorter sticker removal time compared to the ischemia group(both P<0.05).The HomeCage test showed that the ischemia group and treatment group displayed a decrease in activity indicators and increase in immobility time compared to the sham-operation group(all P<0.05);compared to the ischemia group,the treatment group showed significantly increased activity indicators and decreased immobility time(all P<0.05).Western blot showed that penumbral protein levels of BDNF,SYP-1 and GAP-43 were significantly increased 14 days after model establishment,and the increase was more pronounced in the treatment group(all P<0.05);the penumbral NF expression was significantly reduced,and regular whisker stimulation was a

关 键 词：脑梗死 神经可塑性 胡须刺激 神经丝蛋白 生长相关蛋白43 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学] R743