通督调神针法调控MAPK/ERK信号通路减轻大鼠脑缺血再灌注后血脑屏障损伤的机制研究  

Research of Tongdu Tiaoshen Acupuncture Alleviates BBB Damage Mechanism by Regulating the MAPK/ERK Signaling Pathway in Cerebral Ischemia Reperfusion Rats

作　　者：李佳欣 张小卿[1,2] 胡楠 董佳梓 隋月皎 陈怡然[1,2] 江静 孙世婧[1] LI Jiaxin;ZHANG Xiaoqing;HU Nan;DONG Jiazi;SUI Yuejiao;CHEN Yiran;JIANG Jing;SUN Shijing(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Key Laboratory of Acupuncture Biology of Liaoning Province Department of Education,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁省教育厅针灸生物学重点实验室,辽宁沈阳110847 [3]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《辽宁中医药大学学报》2025年第4期139-144,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：辽宁省科技厅应用基础研究计划项目(2023JH2/101300052);国家自然科学基金青年科学基金项目(82305391);辽宁省教育厅针灸生物学重点实验室建设项目(LJ232410162029);辽宁省科技厅应用基础研究计划项目(2023JH2/101700241);辽宁省教育厅高等学校基本科研项目(JYTMS20231820)。

摘　　要：目的探究MAPK/ERK信号通路在通督调神针刺法,干预大鼠脑缺血再灌注后血脑屏障(BBB)损伤中的作用。方法50只SpragueDawley(SD)大鼠,线栓法制备大鼠大脑中动脉短暂性脑缺血再灌注损伤模型(CIRI),缺血2 h,选用Zea-Longa评分评判造模成功与否,将符合标准的纳入实验并随机分为模型组与电针组;假手术组仅剥离颈动脉,不进行栓塞。电针组对大鼠选取“百会”“印堂”和双侧“足三里”进行干预(4 Hz/20 Hz,15 min,1次/d),模型组与假手术组仅予同等时间捆绑。7 d再次对各组大鼠进行Garcia JH评分神经功能缺损评分后处死取材;采用伊文思蓝染色法(Evans Blue,EB)检测各组大鼠BBB损伤程度;采用2,3,5-苯氯化四氮唑(TTC)染色法分析脑梗死面积评估各组大鼠脑损伤程度;苏木素-伊红(HE)染色分析各组大鼠缺血侧海马组织的结构变化;Western blot检测MAPK/ERK信号通路中p-p38 MAPK、p-ERK1/2蛋白含量及ZO-1、Claudin-5、Occludin蛋白含量;酶联免疫吸附试验(Elisa)法检测ZO-1、Claudin-5、Occludin含量。结果与假手术组比较,模型组的大鼠的神经功能缺损评分降低(P<0.05),脑梗死面积显著增加(P<0.01),血脑屏障通透率升高(P<0.01),海马组织结构明显异常,p-p38 MAPK蛋白表达水平明显增加(P<0.01),p-ERK1/2、ZO-1、Claudin-5、Occludin蛋白表达水平明显减少(P<0.01);与模型组比较,电针后脑缺血再灌注损伤大鼠的神经功能缺陷评分明显升高(P<0.01),梗死面积明显减少(P<0.01),血脑屏障通透率降低(P<0.01),海马组织病理程度明显减轻,p-p38 MAPK蛋白表达水平明显减少(P<0.01),p-ERK1/2、ZO-1、Claudin-5、Occludin蛋白表达水平明显增加(P<0.01)。结论电针可通过调节MAPK/ERK信号通路改善脑缺血再灌注大鼠血脑屏障损伤。Objective To explore Tongdu Tiaoshen acupuncture the role of MAPK/ERK signaling pathway in reducing rats the blood-brain barrier(BBB)damage after cerebral ischemia and reperfusion.Methods 50 SpragueDawley(SD)rats were used to prepare a model of middle cerebral artery transient ischemia and ischemia(CIRI)for 2 h.Zea-Longa score was used to evaluate the model success into the experiment and randomly divided into model group and electroacupuncture group;carotid artery was dissected without embolization.For acupuncture group,“Baihui(GV20)”“Yintang(GV29)”and“Zusanli(ST36)”(both sides)were selected(4 Hz/20 Hz,15 min,1 time/1 d),the model group and the sham group were not acupuncture and only tied for the same time.7 d rats in each group were again scored for Garcia JH neurological deficit and then in using materials;The degree of BBB damage in each group was determined by Evans blue staining;adopted the 2,3,5-tetraazolide(TTC)staining to assess the degree of brain damage in each group;Hematoxylin-eosin(HE)staining analyzed the structural changes of the ischemic side of rats;Western blot to determine the content of p-p38 MAPK,p-ERK 1/2 protein and ZO-1,claudin-5 and occludin protein in MAPK/ERK signaling pathway;ZO-1,claudin-5 and occludin were measured by enzyme-linked immunosorbent assay(Elisa).Results Comparison of the sham-surgery group,in rats in the model group,markedly decreased neurological deficit scores(P<0.05),infarct area increased(P<0.01),blood-brain barrier permeability increased(P<0.01),and abnormal hippocampal tissue structure appeared significantly,expression levels of p-p38 MAPK protein were increased(P<0.01),the protein expression levels of p-ERK 1/2,ZO-1,claudin-5,and occludin were significantly reduced(P<0.01);Compared with the model group,CIRI rats after electroacupuncture showed significantly increased neurologic deficit scores(P<0.01),significantly reduced infarct area(P<0.01),decreased blood-brain barrier permeability rate(P<0.01),significantly reduced degree of hippocampal histopath

关 键 词：脑缺血再灌注 电针 MAPK/ERK信号通路 血脑屏障 大鼠 

分 类 号：R245[医药卫生—针灸推拿学]