通督调神针法调控外泌体抑制大脑中动脉闭塞/再通大鼠脑凋亡的机制研究  

Study on the mechanism of regulating exosomes to inhibit brain apoptosis in MCAO/R rats by Tongdu Tiaoshen needles therapy

作　　者：王一鹤 杨家庆 赵忠海[1] 董宝强 隋月皎 杨芳[4] 张小卿 WANG Yihe;YANG Jiaqing;ZHAO Zhonghai;DONG Baoqiang;SUI Yuejiao;YANG Fang;ZHANG Xiaoqing(Department of Rehabilitation Medicine,Central Hospital Affiliated to Shenyang Medical College,Shenyang 110075,China)

机构地区：[1]沈阳医学院附属中心医院康复医学科,110075 [2]辽宁中医药大学针灸推拿学院 [3]辽宁省教育厅针灸生物学重点实验室 [4]辽宁中医药大学中医学院

出　　处：《环球中医药》2025年第4期693-700,共8页Global Traditional Chinese Medicine

基　　金：国家重点研发计划(2019YFC1708701);辽宁省科技厅应用基础研究计划项目(2023JH2/101300052);辽宁省教育厅重点攻关项目(L202065);辽宁省教育厅针灸生物学重点实验室建设项目(LJ232410162029)。

摘　　要：目的 探讨通督调神针法是否通过外泌体途径改善脑缺血再灌注大鼠的运动功能。方法 40只雄性SD大鼠随机划分假手术组10只与手术组30只。手术组以改良线栓法制备大脑中动脉闭塞/再通(middle cerebral artery occlusion/reperfusion, MCAO/R)模型,缺血2小时后再灌注。按Longa 5级神经功能评分进行模型评价,将造模成功的大鼠随机划分为模型组与电针组,每组12只。电针组干预“百会”“印堂”“足三里”,连续7天。7天后,观察各组大鼠步态;脑组织取材进行苏木素—伊红染色,观察皮质海马区病理改变;蛋白免疫印迹法检测脑组织中CD9、CD63、CD81、热休克蛋白70(heat shock protein 70,HSP70)、肿瘤易感基因101(tumor susceptibility gene 101,TSG101)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)及Bcl-2相关X蛋白(Bcl-2 associated X,Bax)的含量。结果 与模型组相比,电针组大鼠神经功能评分降低(P<0.05),步态表现提升,脑组织形态结构改善;CD9、CD63、CD81、HSP70及TSG101的表达量增加(均P<0.05);Bax的表达量降低(P<0.05),Bcl-2的表达量及Bcl-2/Bax比值提高(均P<0.05)。结论 通督调神针法能改善MCAO/R大鼠的运动功能,其机制可能与促进外泌体分泌从而抑制凋亡有关。Objective To investigate whether Tongdu Tiaoshen needles therapy can improve motor function of MCAO/R rats through exosome pathway.Methods 40 male SD rats were randomly divided into sham operation group(10 rats)and operation group(30 rats).In the operation group,a middle cerebral artery occlusion/reperfusion(MCAO/R)model was prepared by modified thrombus method.The model was reperfusion after 2 hours of ischemia.Model evaluation was conducted according to Longa 5 neurological function score,and the successful establishment rats were randomly divided into the model group and the electroacupuncture group,with 12 rats in each group.The electroacupuncture group intervened in Baihui,Yintang and Zusanli for 7 consecutive days.After 7 days,the gait in each group was observed.Hematoxylin-eosin staining was performed on brain samples to observe the pathological changes of cortical hippocampus.The contents of CD9,CD63,CD81,heat shock protein 70(HSP70),tumor susceptibility gene 101(TSG101),B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X(Bax)in brain tissue were detected by Western blot.Results Compared with model group,the nerve function score of rats was decreased(P<0.05),gait performance was improved,and brain structure improved in the electroacupuncture group.The expressions of CD9,CD63,CD81,HSP70 and TSG101 were increased(all P<0.05).The expression of Bax was decreased(P<0.05),while the expression of Bcl-2 and the ratio of Bcl-2 to Bax were increased(both P<0.05).Conclusion Tongdu Tiaoshen therapy can improve the motor function of MCAO/R rats,and its mechanism may be related to promoting exosome secretion and inhibiting apoptosis.

关 键 词：脑缺血再灌注 MCAO/R 电针 外泌体 通督调神针法 凋亡 步态 

分 类 号：R245[医药卫生—针灸推拿学]