高频经颅磁刺激对脑缺血再灌注损伤大鼠海马神经元自噬的影响  

Effects of high-repetitive transcranial magnetic stimulation on autophagy of hippocampal neurons in rats with cerebral ischemia-reperfusion injury

作　　者：贾飞阳 周君 刘丹妮 彭昕珂 黄夏荣 李书枝 罗敷 刘红雅 周桂娟 孙智路 刘静 JIA Fei-Yang;ZHOU Jun;LIU Dan-Ni(Department of Rehabilitation Medicine,The First Affiliated Hospital,Hengyang Medical College of Nanhua University,Hengyang 421001,Hunan,China)

机构地区：[1]南华大学衡阳医学院附属第一医院康复医学科,湖南衡阳421001 [2]南华大学衡阳医学院附属第一医院针灸/康复实验室,湖南衡阳421001 [3]南华大学衡阳医学院附属第一医院急诊科,湖南衡阳421001

出　　处：《中国老年学杂志》2024年第22期5502-5506,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金(82002400);湖南省自然科学基金(2023JJ40589);湖南省研究生创新课题(CX20221001);南华大学附属第一医院4310计划(20214310NHYCG07);南华大学研究生科研创新项目(223YXC025)。

摘　　要：目的探究高频经颅磁刺激(rTMS)对脑缺血再灌注损伤(CIRI)大鼠海马神经元的影响。方法36只雄性SD大鼠随机分为假手术组、模型组、rTMS组,每组12只。造大脑中动脉缺血再灌注(MCAO/R)大鼠模型。24 h后采用Longa神经功能评分判断造模是否成功;rTMS组予以rTMS干预,术后第7天用Longa神经功能评分评估神经缺损程度。苏木素-伊红(HE)染色法观察左侧海马组织病理形态改变;酶联免疫吸附试验(ELISA)检测脑脊液肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-1β含量;实时荧光定量聚合酶链反应(RT-PCR)和Western印迹分别检测缺血区脑组织P62、Beclin-1、VPS34、微管相关蛋白1轻链(LC)3B mRNA和蛋白表达。结果与假手术组比较,模型组Longa神经功能评分、脑脊液IL-6、IL-1β、TNF-α含量及海马组织P62、Beclin-1、VPS34 mRNA及蛋白、LC3B mRNA和LC3B-Ⅱ/Ⅰ明显升高(均P<0.05),HE染色显示神经元排列紊乱,结构不清;与模型组比较,rTMS组以上指标明显降低(均P<0.05),HE染色显示神经元受损程度减轻。结论高频rTMS治疗可能通过抑制海马神经元自噬相关蛋白和减轻炎症反应减轻MCAO/R大鼠脑组织损伤。Objective To investigate the effects of high-frequency transcranial magnetic stimulation(rTMS)on hippocampal neurons in rats subjected to cerebral ischemia-reperfusion injure(CIRI).Methods A total of 36 male SD rats were randomly divided into sham-operated group,model group,and rTMS group,with 12 rats in each group.A model of middle cerebral artery occlusion followed by reperfusion(MCAO/R)was established.24 h post-surgery,the Longa neurological score was employed to assess the success of the model.The rTMS group was given rTMS intervention,and on the seventh postoperative day,the Longa neurological score was used to evaluate the degree of neurological deficit.Hematoxylin-eosin(HE)staining was performed to observe pathological changes in the left hippocampal tissue.Enzyme-linked immunosorbent assay(ELISA)was used to measure the contents of tumor necrosis factor(TNF)-α,interleukin(IL)-6,and IL-1βin the cerebrospinal fluid.Real-time fluorescence polymerase chain reaction(PCR)and Western blotting were conducted to detect the mRNA and protein expressions of P62,Beclin-1,VPS34,and microtubule-associated protein 1 light chain(LC)3B in the ischemic brain tissue,respectively.Results Compared with the sham-operated group,Longa neurological scores,cerebrospinal fluid contents of IL-6,IL-1β,TNF-α,mRNA and proteins of P62,Beclin-1,VPS34 in hippocampal tissue,LC3B mRNA and LC3B-Ⅱ/Ⅰwere significantly increased in the model group(all P<0.05).HE staining revealed disordered neuronal arrangement and unclear structural integrity.Compared with model group,the above indexes in rTMS group were significantly decreased(all P<0.05),with a decrease in the extent of neuronal damage which observed in HE staining.Conclusions High-frequency rTMS may mitigate brain tissue injury in MCAO/R rats by inhibiting autophagy-related proteins in hippocampal neurons and reducing inflammatory responses.

关 键 词：脑缺血再灌注损伤 经颅磁刺激 自噬 神经炎性反应 海马体 

分 类 号：R49[医药卫生—康复医学]