电针干预脑缺血再灌注模型大鼠的神经保护机制  

Neuroprotective mechanism of electroacupuncture in cerebral ischemia-reperfusion model rats

作　　者：吴海洋[1] 段冕 李成龙 张君宇[1] 计海生 王海涛[1] 茅伟 王颖[1] Wu Haiyang;Duan Mian;Li Chenglong;Zhang Junyu;Ji Haisheng;Wang Haitao;Mao Wei;Wang Ying(The Second Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230001,Anhui Province,China;Hefei Second People’s Hospital,Hefei 230001,Anhui Province,China;School of Graduate,Anhui University of Chinese Medicine,Hefei 230000,Anhui Province,China;Department of TCM,Affiliated Brain Hospital of Guangzhou Medical University,Guangzhou 510370,Guangdong Province,China)

机构地区：[1]安徽中医药大学第二附属医院,安徽省合肥市230001 [2]合肥市第二人民医院,安徽省合肥市230001 [3]安徽中医药大学研究生院,安徽省合肥市230000 [4]广州医科大学附属脑科医院中医科,广东省广州市510370

出　　处：《中国组织工程研究》2025年第18期3811-3818,共8页Chinese Journal of Tissue Engineering Research

基　　金：安徽省高等学校科学研究项目(2022AH050503),项目负责人:吴海洋;安徽省高等学校科学研究项目(2023AH040111),项目负责人:王颖;安徽省高等学校科学研究项目(2023AH050816),项目负责人:计海生;安徽省高等学校科学研究项目(2023AH050739),项目负责人:张君宇;安徽省卫生健康委科研项目(AHWJ2023A20030),项目负责人:吴海洋;安徽省名中医王颖工作室建设项目(中发展[2022]5号),项目负责人:王颖;安徽省临床医学研究转化专项项目(202304295107020103),项目负责人:王颖;安徽中医药大学临床科研项目(2021efylc11),项目负责人:吴海洋。

摘　　要：背景:前期研究表明,针刺督脉治疗缺血性脑卒中的疗效确切,可通过减轻细胞焦亡改善脑缺血再灌注损伤,但上游调控机制尚不完全明确。目的:分析电针对脑缺血再灌注损伤模型大鼠的脑保护作用机制。方法:采用随机数字表法将27只SD大鼠随机分为假手术组、模型组和电针组,每组9只。取模型组和电针组大鼠,采用改良线栓法建立脑缺血再灌注损伤模型,验证造模成功后,对电针组大鼠施予电针干预,选取百会、风府、大椎3个穴位,1次/d,20 min/次,连续干预7 d。电针干预结束后,利用神经功能缺损评分、爬杆实验评估大鼠行为学改变,TTC染色评估大鼠脑梗死体积,苏木精-伊红染色观察大鼠梗死侧脑皮质组织形态学变化,免疫荧光染色分析梗死侧脑皮质中Iba-1及活性氧表达,ELISA法检测梗死侧脑皮质组织中白细胞介素1β、白细胞介素6和肿瘤坏死因子α水平,RT-qPCR及Western blot检测大鼠梗死侧脑皮质组织中硫氧还蛋白互作蛋白、NLRP3、Caspase-1、白细胞介素1β的mRNA和蛋白表达,免疫共沉淀法分析大鼠梗死侧脑皮质组织中硫氧还蛋白互作蛋白与NLRP3的相互作用。结果与结论:①与假手术组比较,模型组大鼠神经功能缺损评分、爬杆实验评分及脑梗死体积均升高(P<0.05),Iba-1、活性氧免疫荧光表达增强(P<0.05),白细胞介素1β、白细胞介素6、肿瘤坏死因子α水平均升高(P<0.05),硫氧还蛋白互作蛋白、NLRP3、Caspase-1、白细胞介素1β的mRNA和蛋白表达均升高(P<0.05),苏木精-伊红染色显示模型组大鼠梗死侧脑皮质神经元变性坏死,细胞核出现碎裂溶解及细胞空泡现象;②与模型组比较,电针组大鼠神经功能缺损评分、爬杆实验评分及脑梗死体积均降低(P<0.05),Iba-1、活性氧免疫荧光表达减弱(P<0.05),白细胞介素1β、白细胞介素6、肿瘤坏死因子α水平均降低(P<0.05),硫氧还蛋白互作蛋白、NLRP3、CaspaBACKGROUND:Previous studies have demonstrated that acupuncture at the governor meridian has precise efficacy in the treatment of ischemic stroke and can improve cerebral ischemia-reperfusion injury by attenuating pyroptosis,but the upstream regulatory mechanisms are not yet fully clarified.OBJECTIVE:To observe the neuroprotective effect of electroacupuncture in model rats of cerebral ischemia-reperfusion injury.METHODS:Twenty-seven Sprague-Dawley rats were randomly divided into sham surgery,model,and electroacupuncture groups,with nine rats in each group.Modified suture method was used to establish cerebral ischemia-reperfusion model rats in the model and electroacupuncture groups.The electroacupuncture group was subjected to electroacupuncture at“Baihui,”“Fengfu,”and“Dazhui”acupoints,20 minutes each,once a day,for 7 consecutive days.After treatment,neurological deficit scoring and pole test were performed to assess behavioral changes.Tri-phenyl tetrazolium chloride staining was used to assess cerebral infarction size in rats.Hematoxylin-eosin staining was performed to observe morphological changes in cerebral cortex tissue on the infarcted side of rats.Immunofluorescence analysis was used to determine Iba-1 and reactive oxygen species levels in cerebral cortex tissue on the infarcted side of rats,ELISA method was used for measuring interleukin-1β,interleukin-6 and tumor necrosis factorαlevels in cerebral cortex tissue on the infarcted side of rats.Real-time fluorescence quantitative PCR and western blot were used to detect mRNA and protein expression levels of thioredoxin interaction protein,nod-like receptor associated protein 3(NLRP3),Caspase-1 and interleukin-1βin cerebral cortex tissue on the infarcted side of rats respectively,and the interaction between thioredoxin interaction protein and NLRP3 was analyzed by immunoprecipitation.RESULTS AND CONCLUSION:(1)Compared with the sham surgery group,rats in the model group showed an increase in neurological deficit score,pole test score,cerebral inf

关 键 词：脑缺血再灌注损伤 电针 NLRP3 活性氧 硫氧还蛋白互作蛋白 小胶质细胞 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R245.31