摘要
目的:观察醒脑调肾电针法对创伤后应激障碍(PTSD)大鼠杏仁核、海马区环磷腺苷效应元件结合蛋白(CREB)的表达,及其与突触关键蛋白结合能力的影响,探讨针刺改善PTSD的突触可塑性机制。方法:SD大鼠随机分为空白组、模型组、电针组,每组8只。应用单一连续应激法建立PTSD模型。电针组于"百会""神庭""肾俞"(双侧)穴行电针治疗,每日1次,每次20 min,连续治疗21 d。以自发活动、条件性恐惧反应检测评价行为学改变,运用免疫组织化学法、Western blot法检测杏仁核、海马区CREB的表达,运用染色质免疫沉淀(CHIP)法验证CREB与突触蛋白的结合能力。结果:与空白组比较,模型组自发活动距离减少(P<0.01),条件性恐惧反应检测僵直时间百分比增加(P<0.01);与模型组比较,电针组大鼠自发活动距离增加(P<0.05),条件性恐惧反应检测僵直时间百分比降低(P<0.01,P<0.05)。与空白组比较,模型组大鼠杏仁核、海马脑区的CREB表达水平下降(P<0.01);与模型组比较,电针组大鼠杏仁核、海马区CREB蛋白表达明显提高(P<0.01)。与空白组比较,模型组杏仁核、海马区CREB与突触后致密蛋白95(PSD95)结合能力下降(P<0.01),与突触素(SYN)、生长相关蛋白43(GAP43)结合能力差异无统计学意义(P>0.05);与模型组比较,电针组杏仁核、海马区CREB与PSD95蛋白结合能力提高(P<0.05),与SYN、GAP43结合能力无显著变化(P>0.05)。结论:电针可能是通过提高CREB与突触关键蛋白PSD95结合能力以调控PTSD模型大鼠杏仁核、海马脑区突触可塑性,进而改善PTSD大鼠的行为学表现。
Objective To observe the effect of electroacupuncture(EA)on the expression of cAMP-response element binding protein(CREB,a key protein for BDNF-TrkB signaling)and it’s blinding ability to synaptic key protein in the amygdala and hippocampus of rats with post-traumatic stress disorder(PTSD),so as to lay a foundation for further study of the interaction mechanism between BDNF-TrkB signaling and synaptic plasticity.Methods Twenty-four male SD rats were randomly divided into blank,model and electroacupuncture(EA)groups,with 8 rats in each group.The PTSD model was established by psychological stress(bondage)and physiological stress(forced swimming and anesthesia).After modeling,EA(2 Hz/100 Hz,1 mA)was applied to"Baihui"(GV20)"Shenting"(GB24)and bilateral"Shenshu"(BL23)for 20 min,once daily for 21 days.The behavioral changes(spontaneous locomotor within 30 min and contextual fear conditioning tests in 7 days)were detected by using a spontaneous locomotor detection box,and a conditioned fear response test chamber,respectively.The expression of CREB was detected by immunohistochemistry and Western blot,separately.The binding abilities of CREB to synaptic proteins(post synaptic density 95[PSD95],synaptophysin[SYN]and growth-associated protein 43[GAP43])were verified by chromatin-immunoprecipitation(CHIP)technique.Results After modeling,the spontaneous locomotor distance,the expression levels of CREB and the binding ability of CREB to PSD95 protein in the amygdala and hippocampus were significantly decreased(P<0.01),and the percentage of freezing time significantly increased in the model group relevant to the blank group(P<0.01).Following the intervention,the spontaneous locomotor distance,and the expression levels of CREB and the binding ability of CREB to PSD95 protein were considerably increased in the EA group relevant to the model group(P<0.05,P<0.01).No significant changes were found in the binding abilities of CREB to SYN and GAP43 after modeling and after EA intervention(P>0.05).Conclusion EA can improve the motor activity in PTSD rats,which may be associated with its effect in increasing the binding ability of CREB to the synaptic key protein PSD95 to regulate the interaction between the synaptic plasticity and BDNF-TrkB signaling pathway of the amygdala and hippocampus.
作者
李秘
李凯
丁宁
谢毅强
牛坤
张虹
LI Mi;LI Kai;DING Ning;XIE Yi-qiang;NIU Kun;ZHANG Hong(School of Traditional Chinese Medicine,Hainan Medical University,Haikou 571199,China;Clinical Medical College,Chengdu University of Traditional Chinese Medicine,Chengdu 610075;College of Acupuncture-moxibustion and Tuina,Chengdu University of Traditional Chinese Medicine,Chengdu 610075)
出处
《针刺研究》
CAS
CSCD
北大核心
2020年第7期517-523,共7页
Acupuncture Research
基金
国家自然科学基金面上项目(No.81574086)
海南省自然科学基金青年项目(No.819QN223)。
关键词
电针
创伤后应激障碍
杏仁核
海马
环磷腺苷效应元件结合蛋白
结合能力
Electroacupuncture
Post-traumatic stress disorder
Amygdala
Hippocampus
cAMP-response element binding protein(CREB)
Binding ability
