心肌缺血小鼠背根节神经元兴奋性改变与穴位敏化的关系研究  

Relationship between acupoint sensitization and changes in dorsal root ganglion neuron excitability in myocardial ischemia mice

作　　者：胡瑞斌 龙梦阳 刁志君 吴宇蔚 薛思梦 高昕妍[4] 乔海法 HU Rui-bin;LONG Meng-yang;DIAO Zhi-jun;WU Yu-wei;XUE Si-meng;GAO Xin-yan;QIAO Hai-fa(College of Acupuncture-moxibustion and Tuina,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Shaanxi Key Laboratory of Acupuncture and Medicine,Xianyang 712046,Shaanxi Province;Traditional Chinese Medicine Master Research Institute,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province;Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,Beijing 100700;Xianyang Key Laboratory of Neurobiology(Acupuncture),Xianyang 712046,Shaanxi Province)

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]陕西中医药大学国医大师研究院,陕西咸阳712046 [4]中国中医科学院针灸研究所,北京100700 [5]咸阳市神经生物学(针灸)重点实验室,陕西咸阳712046

出　　处：《针刺研究》2023年第9期833-842,共10页Acupuncture Research

基　　金：国家自然科学基金项目(No.81873385、82074557);国家重点研发计划项目(No.2018YFC1704606)。

摘　　要：目的:探讨心肌缺血(MI)模型小鼠体表穴位敏化状态与其所对应脊髓节段背根神经节(DRG神经元电生理特性改变的关系。方法:将68只雄性C57BL/6J小鼠随机分为对照组和模型组,每组34只。模型组以160 mg/kg的异丙肾上腺素(ISO)腹腔注射建立MI模型,对照组注射相同剂量的0.9%氯化钠溶液。造模后第6天,苏木精-伊红(HE)染色法观察小鼠心脏组织病理形态变化;对小鼠尾静脉注射伊文思蓝(EB)染色液,观察体表渗出点的部位、分布和数量;离体电生理实验评估不同类型的DRG神经元全细胞跨膜总电流、内在兴奋性及激活膜特性等电生理特性。结果:与对照组比较,模型组小鼠心脏明显变大,HE染色后镜下观察心肌肥厚增大,细胞结构破坏,心肌细胞间有单核细胞浸润等病理表现;模型组EB渗出点数量显著增多(P<0.01),且主要集中在脊髓胸(T)1—T5节段邻近表皮、“肺俞”“厥阴俞”及“心俞”等部位;模型组DRG中型神经元全细胞跨膜总电流显著增大,放电数目、平均瞬时频率、平均放电频率显著升高(P<0.01),基强度、动作电位幅值显著降低(P<0.01,P<0.05);模型组DRG小型神经元膜特性及内在兴奋性无显著性改变。模型组DRG中降钙素基因相关肽阳性表达,且被生物素标记的DRG中型神经元跨膜总电流显著增大,放电数目和平均瞬时频率均显著增加(P<0.05,P<0.01),基强度显著降低(P<0.05)。结论:ISO诱导小鼠MI模型,可能是通过T1—T5节段的DRG中型神经元膜特性和内在兴奋性改变介导体表穴位敏化的发生。Objective To investigate the relationship between the sensitization state of acupoints on the sur⁃face of the myocardial ischemia(MI)model mice and the changes in the electrophysiological properties of the dorsal root ganglion(DRG)neurons in the corresponding spinal cord segment,and its underlying mechanism.Methods Sixty-eight male C57BL/6J mice were randomly divided into control and model groups(34 mice in each group).The model group received an intraperitoneal injection of 160 mg/kg isoproterenol(ISO)to establish the MI model,and the control group received an injection of the same dose of normal saline as the model group.After modeling for about 6 days,MI proportion was measured by HE staining to verify the pathological changes in the heart tissue.Evans blue(EB)dye was injected into the tail vein of mice to reflect the size,location,distribution,and number of exudates on the body surface.Then,whole-cell membrane currents,intrinsic excitability and membrane properties of different types of DRG neurons were evaluated by electrophysiological experiment in vitro.Results Compared with the control group,the heart size was larger,with pathological outcomes showing enlarged myocardial hypertrophy,destroyed structure of cardiomyocytes,with mononuclear cell infiltration among the cardiomyocytes in the model group.Compared with the control group,the number of EB exudation points was significantly increased(P<0.01),which were mainly concen⁃trated in the epidermis near the T1—T5 segment of the spinal cord,“Feishu”(BL13),“Jueyinshu”(BL14)and“Xin⁃shu”(BL15)in the model group.Compared with the control group,the rheobase and action potential amplitude(APA)of DRG medium-sized neurons were obviously decreased(P<0.01,P<0.05),while the whole-cell membrane currents,the spike numbers,the average instantaneous frequency,and the average discharge frequency were markedly in⁃creased(P<0.01).There were no significant alterations in the membrane properties and intrinsic excitability induced by depolarized currents of

关 键 词：心肌缺血 穴位敏化 背根神经节 神经元内在兴奋性 

分 类 号：R245-0[医药卫生—针灸推拿学]