前扣带回皮层中CaMKⅡα^(+)GAD67^(+)神经元在大鼠神经病理性疼痛和焦虑抑郁共病中的作用  

Effect of CaMKⅡα^(+)and GAD67^(+)neurons in the anterior cingulate cortex in the co-morbidity of neuropathic pain and anxiety,depression in rats

作　　者：王文婷 许耀威 白倩[1] 李治松 WANG Wenting;XU Yaowei;BAI Qian;LI Zhisong(The Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450003,China;Institute of Neuroscience,Academy of Medical Sciences,Zhengzhou University,Zhengzhou 450000,China;The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第二附属医院,河南郑州450003 [2]郑州大学医学科学院神经科学研究所,河南郑州450000 [3]郑州大学第一附属医院,河南郑州450052

出　　处：《中国实用神经疾病杂志》2024年第1期1-8,共8页Chinese Journal of Practical Nervous Diseases

基　　金：河南省医学科技攻关计划省部共建青年项目(编号:SBGJ2021030880);河南省中青年卫生科技创新优秀青年人才培养项目(编号:YXKC2020059,YXKC2021018)。

摘　　要：目的探讨前扣带回皮层(ACC)中表达钙/钙调素依赖性蛋白激酶Ⅱ(CaMKⅡ)的兴奋性神经元和表达谷氨酸脱羧酶67(GAD67)的抑制性神经元在神经病理性疼痛以及焦虑、抑郁共病中的作用。方法雄性SD大鼠72只,随机分为假手术组(Sham)、神经病理性疼痛组(CCI)、神经病理性疼痛+CaMKⅡα非激活组(CCI+hM4Di^(-))、神经病理性疼痛+CaMKⅡα激活组(CCI+hM4Di^(+))、神经病理性疼痛+GAD67非激活组(CCI+hM3Dq^(-))、神经病理性疼痛+GAD67激活组(CCI+hM3Dq^(+))。通过由专门设计药物技术激活的设计受体(DREADDs)偶联hM3Dq或hM4Di,以细胞类型和时间依赖性的方式调控神经元活性。以机械缩足阈值(PWT)、热缩足潜伏期(PWL)评估疼痛行为,旷场试验、新环境进食抑制试验、强迫游泳试验评估焦虑、抑郁样行为,WesternBlot检测CaMKⅡα和GAD67表达,c-Fos免疫荧光染色检测神经元激活情况。结果与Sham组相比,CCI组大鼠术后表现出机械痛觉过敏和热痛觉超敏反应以及焦虑、抑郁样行为;对侧ACC中CaMKⅡα蛋白表达水平升高,GAD67蛋白表达水平降低;免疫荧光染色显示CaMKⅡα+神经元激活增多,GAD67^(+)神经元激活减少。与CCI+hM4Di^(-)组大鼠相比,CCI+hM4Di^(+)组在术后28 dPWL、PWT升高,焦虑和抑郁样行为改善。与CCI+hM3Dq^(-)组相比,CCI+hM3Dq^(+)组在术后28dPWL、PWT升高,焦虑、抑郁样行为减少。结论抑制ACC中CaMKⅡα+兴奋性神经元或激活GAD67^(+)抑制性神经元能起到减轻NP大鼠疼痛,改善焦虑、抑郁样行为的作用。Objective To investigate the role of excitatory neurons expressing calcium/calmodulin-dependent protein kinaseⅡ(CaMKⅡ)and inhibitory neurons expressing glutamic acid decarboxylase 67(GAD67)in the anterior cingulate cortex(ACC)in neuropathic pain as well as anxiety and depression co-morbidity.Methods Seventy-two male SD rats were randomly divided into sham-operated group(Sham),neuropathic pain group(CCI),neuropathic pain+CaMKⅡαinactivation group(CCI+hM4Di^(-)),neuropathic pain+CaMKⅡαactivation group(CCI+hM4Di^(+)),neuropathic pain+GAD67 inactivation group(CCI+hM3Dq^(-)),and neuropathic pain+GAD67 activation group(CCI+hM3Dq^(+)).Neuronal activity was modulated in a cell-type and time-dependent manner by coupling hM3Dq or hM4Di via designer receptors(DREADDs)activated by a specifically designed drug technique.PWL and PWT were observed in each group of rats.Anxiety and depression-like behaviors were assessed in the absentee field assay,new environment feeding inhibition assay and forced swimming assay.The expressions of CaMKⅡαand GAD67 were detected by Western Blot.The neuronal activation was detected by immunofluorescence staining with c-Fos co-labeling.Results Compared with the Sham group,the CCI rats showed mechanical nociceptive hypersensitivity and thermal hyperalgesia,as well as anxiety and depression-like behaviors.The expression level of CaMKⅡαprotein was increased and that of GAD67 protein was decreased in the contralateral ACC.Immunofluorescence staining showed increased activation of CaMKⅡα^(+)neurons and decreased activation of GAD67^(+)neurons.Compared with the CCI+hM4Di^(-)group,the rats in the CCI+hM4Di+group showed significantly higher PWL and PWT and improved anxiety and depression-like behaviors at 28-day.Compared with the CCI+hM3Dq^(-)group,the CCI+hM3Dq+group showed significantly higher PWL and PWT,and reduced anxiety and depression-like behaviors at 28-day.Conclusion Inhibition of CaMKⅡα^(+)excitatory neurons or activation of GAD67^(+)inhibitory neurons in ACC can reduce pai

关 键 词：神经病理性疼痛 焦虑 抑郁 由特定药物激活的受体 CaMKⅡα^(+)神经元 GAD67^(+)神经元 前扣带回皮层 大鼠 

分 类 号：R-332[医药卫生]