电针联合丹参酮ⅡA经Hippo信号通路改善脑缺血再灌注损伤大鼠神经细胞凋亡的机制研究  

Mechanism of electroacupuncture combined with tanshinoneⅡA in improvement of neuronal apoptosis in rats with cerebral ischemia-reperfusion injury via Hippo signaling pathway

作　　者：闫佳兰 高利涛 甄宁 王丽 YAN Jialan;GAO Litao;ZHEN Ning;WANG Li(Department of Neurology,the 980th Hospital of Joint Logistic Support Force of the Chinese People′s Liberation Army,Shijiazhuang,Hebei,050082)

机构地区：[1]中国人民解放军联勤保障部队第九八〇医院神经内科,河北石家庄050082

出　　处：《实用临床医药杂志》2025年第8期58-64,共7页Journal of Clinical Medicine in Practice

基　　金：河北省重点研发计划项目(2163820015D)。

摘　　要：目的探讨电针(EA)联合丹参酮ⅡA(TanⅡA)对脑缺血再灌注(I/R)损伤大鼠的神经保护作用及相关机制。方法采用改良Longa线栓法制备脑I/R大鼠模型,并将其随机分为模型组(Model组)、电针组(EA组)、TanⅡA组和针药组(EA+TanⅡA组),每组16只。Sham组(n=16)大鼠仅暴露并游离颈动脉。采用神经功能缺损评分和Morris水迷宫测试评估大鼠的神经功能损伤和认知功能。采用2,3,5-氯化三苯基四氮唑(TTC)染色评估脑梗死体积。采用苏木精-伊红(HE)染色和TUNEL染色评估海马组织神经元损伤。采用蛋白免疫印迹检测凋亡相关蛋白和Hippo信号通路相关蛋白。结果与Sham组比较,Model组大鼠神经功能缺损评分、脑梗死面积百分比、获得性训练期间的逃避潜伏期,以及脑组织TUNEL阳性染色细胞比例、Caspase-3活性、Cleaved Caspase-3蛋白和Bax蛋白表达均升高,而探索训练期间的目标象限停留时间和60 s内穿过平台次数,以及Bcl-2蛋白、Yes相关蛋白(YAP)和含PDZ结合基序的转录共激活因子(TAZ)蛋白表达均降低,差异有统计学意义(P<0.05)。与Model组比较,EA组、TanⅡA组和EA+TanⅡA组神经功能缺损评分、脑梗死体积百分比、获得性训练期间的逃避潜伏期、探索训练期间的目标象限停留时间和60 s内穿过平台次数,以及脑组织TUNEL阳性染色细胞比例、Caspase-3活性、Cleaved Caspase-3蛋白和Bax蛋白表达均降低,且EA+TanⅡA组低于EA组和TanⅡA组,差异均有统计学意义(P<0.05)。与Model组比较,EA组、TanⅡA组和EA+TanⅡA组Bcl-2、YAP和TAZ蛋白表达增加,且EA+TanⅡA组蛋白表达水平高于EA组和TanⅡA组,差异有统计学意义(P<0.05)。结论电针与TanⅡA联合治疗脑I/R损伤能够协同增效,其机制可能与抑制Hippo通路活性有关。Objective To explore the neuroprotective effect and related mechanism of electroacupuncture(EA)combined with tanshinoneⅡA(TanⅡA)for the treatment of rats with cerebral ischemia-reperfusion(I/R)injury.Methods Model rats with cerebral I/R were established by the modified Longa suture occlusion method and randomly divided into Model group,EA group,TanⅡA group,and EA+TanⅡA group,with 16 rats in each group.Rats in the Sham group(n=16)had their carotid arteries exposed and dissected but not occluded.Neurological deficit score and Morris water maze test were used to assess neurological impairment and cognitive function in rats.2,3,5-triphenyltetrazolium chloride(TTC)staining was used to evaluate cerebral infarction volume.Hematoxylin-eosin(HE)staining and TUNEL staining were used to assess neuronal damage in the hippocampus.Western blot was used to detect apoptosis-related proteins and proteins related to the Hippo signaling pathway.Results Compared with the Sham group,the Model group showed increased neurological deficit score,percentage of cerebral infarction area,escape latencies during acquisition training,proportion of TUNEL-positive stained cells in brain tissue,Caspase-3 activity,expression of Cleaved Caspase-3 protein and Bax protein,and decreased expression of time spent in the target quadrant during probe training,the number of platform crossings within 60 s,Bcl-2 protein,Yes-associated protein(YAP),and transcriptional co-activator with PDZ-binding motif(TAZ)protein,and all the differences were significant(P<0.05).Compared with the Model group,the EA group,TanⅡA group,and EA+TanⅡA group showed decreased neurological deficit score,percentage of cerebral infarction volume,escape latencies during acquisition training,time spent in the target quadrant during probe training,number of platform crossings within 60 s,proportion of TUNEL-positive stained cells in brain tissue,Caspase-3 activity,expression of Cleaved Caspase-3 protein and Bax protein,with the EA+TanⅡA group showing lower values than the EA

关 键 词：大鼠 脑缺血再灌注 神经细胞凋亡 电针 丹参酮ⅡA Yes相关蛋白 含PDZ结合基序的转录共激活因子 信号通路 

分 类 号：R741.05[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学] R245.97