电针耳甲区耳穴对创伤后应激障碍小鼠前扣带回皮层谷氨酸能神经元兴奋性水平的影响  

Effects of electroacupuncture at auricular points on excitability of glutamatergic neurons in the anterior cingulate cortex of posttraumatic stress disorder mice

作　　者：左燕 刘永斌 袁子焉 杜瑞冰 吴宇蔚 薛思梦 刁志君 乔海法 ZUO Yan;LIU Yong-bin;YUAN Zi-yan;DU Rui-bing;WU Yu-wei;XUE Si-meng;DIAO Zhi-jun;QIAO Hai-fa(College of Acupuncture-moxibustion and Tuina of Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Shaanxi Key Laboratory of Acupuncture and Medicine,Xianyang 712046,Shaanxi Province,China;Key Laboratory of Acupuncture and Neurobiology,Shaanxi Administration of Traditional Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Traditional Chinese Medicine Master Research Institute,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China)

机构地区：[1]陕西中医药大学针灸推拿学院,陕西咸阳712046 [2]陕西省针药结合重点实验室,陕西咸阳712046 [3]陕西省中医药管理局针灸神经生物学重点实验室,陕西咸阳712046 [4]陕西中医药大学国医大师研究院,陕西咸阳712046

出　　处：《针刺研究》2024年第11期1121-1128,共8页Acupuncture Research

基　　金：陕西省教育厅重点科研计划项目(No.22JS008);国家自然科学基金项目(No.82205266、82074557);陕西中医药大学科技创新人才体系建设计划项目(No.2023-KJXX-03);陕西省自然科学基础研究计划一般项目(No.2019-JM-399)。

摘　　要：目的:从神经元内在兴奋性和神经元功能可塑性的角度探讨耳甲区耳穴与大脑前扣带回皮层(ACC)谷氨酸能神经元交流对话的生物学机制,为阐明针刺耳穴治疗情志类疾病的神经机制提供实验证据。方法:C57BL/6J雄性小鼠随机分为对照组、模型组、假电针组、电针组,每组5只。向小鼠双侧ACC注射AAV2/9-CaMKⅡα-EGFP病毒溶液标记谷氨酸能神经元。使用单程长时应激刺激与足底电击相结合的方法构建创伤后应激障碍小鼠模型。造模1周后,电针组小鼠麻醉后于双侧耳甲区耳穴“心”和耳甲艇中心部位进行电针刺激,每天30 min,连续7 d;假电针组小鼠给予同样麻醉,但不进行电针干预。造模2周后,以旷场实验、高架十字迷宫实验评估小鼠焦虑样行为表现;全细胞膜片钳技术记录ACC谷氨酸能神经元内在兴奋性水平的改变。结果:与对照组比较,模型组小鼠旷场中心区域停留时间、移动距离及高架十字迷宫开臂停留时间百分比、进入开臂次数百分比均减少(P<0.001,P<0.01),ACC谷氨酸能神经元基强度升高(P<0.01),放电数目减少(P<0.05,P<0.01,P<0.001)。与模型组比较,电针组小鼠旷场中心区域停留时间、移动距离及高架十字迷宫开臂的停留时间百分比、进入开臂次数百分比均增加(P<0.001,P<0.01),ACC谷氨酸能神经元基强度降低(P<0.05),放电数目增加(P<0.05)。与假电针组比较,电针组小鼠旷场中心区域停留时间、移动距离及高架十字迷宫开臂的停留时间百分比、进入开臂次数百分比均增加(P<0.001,P<0.01),ACC谷氨酸能神经元基强度降低(P<0.01),放电数目增加(P<0.05)。结论:电针耳甲区耳穴可有效减轻创伤后应激障碍小鼠焦虑样行为,其机制可能是通过提高由创伤性应激经历减弱的ACC谷氨酸能神经元内在兴奋性水平实现的。Objective To elucidate the relationship between the auricular point stimulation and the activity of glutamatergic neurons in the anterior cingulate cortex(ACC)from the perspective of intrinsic excitability plasticity of neurons in mice with posttraumatic stress disorder(PTSD),so as to explore the underlying mechanisms of acupuncture at auricular points in improving emotional diseases induced by PTSD.Methods C57BL/6J male mice were randomly divided into control,PTSD model,sham electroacupuncture(EA),and EA groups,with 5 mice in each group.The glutamatergic neurons were labelled by injection of AAV2/9-CaMKⅡα-EGFP viral fluid into the bilateral ACC.Fourteen days after the injection,the PTSD model was established by single prolonged stress(restraint stress,forced swimming,ether exposure)and plantar electrical shock.EA(2 Hz/15 Hz,1 mA)was applied to bilateral“Xin”points and the center of auriculae of auricular concha area for 30 min,once daily for 7 days.Mice of the sham EA group were anesthetized for 30 min per day for 7 days,but no EA treatments were given.The anxiety-like behavior of mice was evaluated by open field(OF)and elevated plus maze(EPM)tests after 2 weeks of modeling.The whole-cell patchclamp method was used to record the intrinsic excitability level of ACC glutamatergic neurons.Results Compared with the control group,the dwell time and locomotor distance in the central area of the OF,the dwell time and entry times percentage in the open arms of the EPM in the model group were significantly reduced(P<0.001,P<0.01).At the same time,the intrinsic excitability of ACC glutamatergic neurons in model mice was suppressed,presenting as enhanced rheobase currents(P<0.01)and decreased spike number(P<0.05,P<0.01,P<0.001).Compared with the model group,the dwell time and locomotor distance in the central area of the OF,the dwell time and entry times percentage in the open arms of the elevated plus maze were significantly increased(P<0.001,P<0.01),and the intrinsic excitability of ACC glutamatergic neurons was s

关 键 词：电针 耳穴 创伤后应激障碍 前扣带回皮层 神经元内在兴奋性 

分 类 号：R245[医药卫生—针灸推拿学]