摘要
目的观察电针耳甲区对改善急性应激障碍小鼠抑郁样行为及腹侧背盖区(Ventral tegmental area,VTA)中TH、DRD1、DRD2表达的影响。方法实验1:将18只C57雄性小鼠分为对照组9只、模型组9只,第5天以及第36天开始利用强迫游泳和悬尾等行为学范式观察小鼠抑郁样行为;利用免疫荧光染色法检测小鼠VTA中即刻早期基因蛋白(C-Fos)和酪氨酸羟化酶(Tyrosine hydroxylase,TH)双标染色。实验2:将45只雄性C57小鼠,分为空白组、模型组、电针组每组各15只,电针干预完成30天后,利用强迫游泳和悬尾等行为学范式检测小鼠抑郁样行为;通过荧光定量PCR和蛋白免疫印迹技术检测各组小鼠VTA中的多巴胺受体D1(Dopamine receptor D1,DRD1)、多巴胺受体D2(Dopamine receptor D2,DRD2)的表达水平;利用免疫荧光染色法观察小鼠VTA脑区TH神经元数量。结果实验1:与对照组相比,模型组在第5、6天,悬尾(P<0.001)和强迫游泳(P<0.001)的不动时间和静止时间增加,模型组在第36、37天,悬尾(P<0.01)和强迫游泳(P<0.01)的不动时间和静止时间增加;与对照组相比,模型组C-Fos和TH荧光双标神经元数量占比增加(P<0.05)。实验2:电针干预完成30天后,与模型组相比,电针组悬尾(P<0.01)和强迫游泳(P<0.01)的不动时间和静止时间都减少;与模型组相比,电针组VTA中DRD1 mRNA(P<0.001)和蛋白表达(P<0.01)增加,DRD2mRNA(P<0.001)和蛋白表达(P<0.05)减少,TH蛋白表达(P<0.05)增加,TH神经元数量增加(P<0.01)。结论电针耳甲区的早期干预对急性应激障碍小鼠诱发的抑郁样行为有改善作用,可能是通过调控VTA中多巴胺能神经元活动水平以及DRD1、DRD2表达水平实现的。
Objective To observe the effects of electroacupuncture auricular acupuncture points on depression-like behaviors and ventral dorsal tegmental area(VTA)TH,DRD1 and DRD2 induced in well acute stress disorder(ASD)mice.Methods Experiment 1:18 male C57 mice were divided into 9 control and 9 model mice.Behavioural paradigms such as forced swimming test(FST)and tail suspension test(TST)were used to detect depression-like behaviors in the mice starting on day 5 and day 36;double-labelled staining for immediate early gene protein(C-Fos)and tyrosine hydroxylase(TH)was detected in the VTA of the mice by immunofluorescence.Experiment 2:45 male C57 mice were divided into 15 mice each in the control groups,model groups and electroacupuncture groups.30 days after completion of the electroacupuncture intervention,the mice were tested for depressive behaviors using behavioural paradigms such as FST and TST;dopamine receptor D1(DRD1)and dopamine receptor D2(DRD2)was detected in the VTA of mice in each group using fluorescence quantitative PCR and Western blot techniques;the number of TH neurons in the brain area of the VTA of mice was observed by immunofluorescence.Results Experiment 1:Compared with the control group,the immobile duration of the model group increased in the TST(P<0.001)and FST(P<0.001)on day 5 and day 6,and the immobile duration of the model group increased in the TST(P<0.01)and FST(P<0.01)on day 36 and day 37;compared with the control group,the number of C-Fos and TH fluorescence double-labelled neurons in the model group increased(P<0.05).Experiment 2:At day after completion of the electroacupuncture intervention,the immobile duration of TST(P<0.01)and FST(P<0.01)were decreased in the electroacupuncture group compared with the model group;compared with the control group,DRD1 mRNA(P<0.001)and protein expression(P<0.01)were increased in the VTA of the electroacupuncture group,and DRD2 mRNA(P<0.001)and protein expression(P<0.05)were decreased,TH protein expression(P<0.05)increased,and the number of TH neurons increased(P<0.01).Conclusion Early intervention with auricular electroacupuncture points has an ameliorative effect on depression-like behaviors induced by acute stress disorder in mice,which may be achieved by modulating the activity level of dopaminergic neurons and the expression level of DRD1 and DRD2 in the VTA.
作者
张轩宇
韩静
林丽
时钰甦
施琳玉
李永丰
ZHANG Xuanyu;HAN Jing;LIN Li;SHI Yusu;SHI Linyu;LI Yongfeng(College of Acupuncture-moxibustion and Tuina,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Key Laboratory of Modern Teaching Technology,Ministry of Education,Shaanxi Normal University,Xi'an 710062,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第5期1267-1276,共10页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
咸阳市重点研发计划课题(S2024-ZDYF-SF-3089):针刺耳甲区改善急性应激障碍导致的抑郁样情绪作用机制研究,负责人:李永丰
陕西省教育科学规划领导小组办公室教育科学规划课题(SGH23Y2416):探究针灸推拿学研究生的创新思维与科学研究方法的培养与实践策略,负责人:李永丰
陕西省科学技术厅一般项目(青年)(2022JQ-949):针刺通过调节MRN-VH的5-HT能投射改善不可控应激诱发的焦虑情绪机制研究,负责人:李永丰。
关键词
电针耳甲区
急性应激障碍
抑郁样行为
酪氨酸羟化酶
腹侧背盖区
多巴胺系统
Electroacupuncture at auricular points
Acute stress disorder
Depression-like behaviors
Tyrosine hydroxylase
Ventral tegmental area
Dopamine system
