摘要
目的:探讨母婴分离(MS)联合慢性束缚应激(RS)致抑郁小鼠的证候属性及其机制研究。方法:仔鼠出生第0天(PD0)随机分为空白组和造模组。PD21后剔除雌鼠,随机分为模型组、温阳组、解郁组、温阳解郁组、氟西汀组,采用MS+RS建立二次打击模型,每组15只。采用糖水、悬尾及旷场实验评估小鼠焦虑抑郁样行为;酶联免疫吸附测定法(ELISA)检测各组小鼠血浆促肾上腺皮质激素(ACTH)及皮质酮(CORT)含量;高效液相-电化学法(HPLC-ECD)检测各组小鼠海马单胺类神经递质含量;实时荧光定量聚合酶链式反应(Real-time PCR)检测各组小鼠海马5-羟色胺(5-HT)系统、下丘脑-垂体-肾上腺(HPA)轴及脑源性神经营养因子(BDNF)信号通路基因表达水平;免疫组化法(IHC)检测各组小鼠海马5-HT系统及HPA轴蛋白表达水平;全自动蛋白表达分析系统(Simple Wes)检测各组小鼠海马BDNF等蛋白表达水平。结果:与空白组比较,模型组小鼠出现抑郁样行为,温阳解郁方及氟西汀可显著缓解。与空白组比较,模型组血浆CORT及ACTH含量显著升高(P<0.01);与模型组比较,温阳解郁方及氟西汀组小鼠血浆CORT及ACTH含量明显降低(P<0.05,P<0.01)。与空白组比较,单胺类神经递质表达方面,模型组海马神经递质表达水平明显降低(P<0.05,P<0.01);与模型组比较,温阳解郁方及氟西汀组小鼠海马神经递质表达水平明显升高(P<0.05,P<0.01)。与空白组比较,模型组小鼠5-HT能神经受到抑制、HPA轴异常激活,温阳解郁方及氟西汀可调控5-HT能神经及HPA轴的异常状态。与空白组比较,海马组织BDNF、TrkB mRNA及蛋白表达明显降低(P<0.05,P<0.01);与模型组比较,温阳、解郁、温阳解郁方及氟西汀组小鼠海马BDNF、TrkB mRNA及蛋白表达水平明显升高(P<0.05,P<0.01)。结论:二次打击致抑郁小鼠的证候属性可能是阳虚肝郁型,温阳解郁方可能通过调节5-HT神经系统及HPA轴介导的BDNF信号通路,增加海马神经递质含量,进而缓解模型小鼠抑郁样行为。
Objective:To explore the syndromes and mechanisms of depression induced by maternal separation(MS)combined with chronic restraint stress(RS)in mice.Method:On postnatal day 0(PD0),the offspring mice were randomized into a blank group(NC)and a modeling group.The mouse model of depression was established by MS+RS for 21 days.After removal of female mice on PD21,the modeled mice were randomized into model,Wenyang,Jieyu,Wenyang Jieyu,and fluoxetine groups,with 15 mice in each group.The sucrose preference,tail suspension,and open field tests were carried out to evaluate the anxiety and depression-like behavior in mice.Enzyme-linked immunosorbent assay was used to measure the adrenocorticotrophic hormone(ACTH)and corticosterone(CORT)levels in mouse plasma.High performance liquid chromatography-electrochemical detector was used to determine the content of monoamine neurotransmitters in the hippocampus.Real-time fluorescence quantitative polymerase chain reaction was employed to determine the mRNA levels of genes in the 5-hydroxytryptamine(5-HT)system,hypothalamicpituitary-adrenal(HPA)axis,and brain-derived neurotrophic factor(BDNF)signaling pathway in the hippocampus.Immunohistochemistry was employed to determine the expression levels of proteins in the 5-HT system and HPA axis in the hippocampus.The Simple Western system was used to determine the protein levels of BDNF and tyrosine kinase receptor B(TrkB)in the hippocampus.Result:Compared with the NC group,the model group exhibited depression-like behavior,which was significantly relieved by Wenyang Jieyu prescription and fluoxetine.Compared with the NC group,the model group showed elevated levels of CORT and ACTH in the plasma(P<0.01),which,however,were lowered by Wenyang Jieyu prescription and fluoxetine(P<0.05,P<0.01).Compared with the NC group,the model group showed inhibited expression of neurotransmitters in the hippocampus(P<0.05,P<0.01),while Wenyang Jieyu prescription and fluoxetine restored the expression of neurotransmitters(P<0.05,P<0.01).Compared with NC group,the model group showed inhibition of the 5-HTergic nerve and abnormal activation of the HPA axis,and Wenyang Jieyu prescription and fluoxetine regulated the abnormal state of the 5-HTergic nerve and HPA axis.Compared with NC group,the modeling down-regulated the mRNA and protein levels of BDNF and TrkB in the hippocampus(P<0.05,P<0.01),which,however,were recovered in Wenyang,Jieyu,Wenyang Jieyu,and fluoxetine groups(P<0.05,P<0.01).Conclusion:The mouse model of depression induced by MS+RS may present the syndrome of Yang deficiency and liver depression.Wenyang Jieyu prescription may increase the content of hippocampal neurotransmitters by regulating the 5-HT system and the BDNF signaling pathway mediated by the HPA axis,thereby alleviating depression-like behavior in mice.
作者
巩子汉
王英
杨婧雯
梁文青
孟丹华
佘楷杰
梁媛
岳广欣
GONG Zihan;WANG Ying;YANG Jingwen;LIANG Wenqing;MENG Danhua;SHE Kaijie;LIANG Yuan;YUE Guangxin(Ningxia Chinese Medicine Research Center,Yinchuan 750021,China;Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;Traditional Chinese Medicine(TCM)School,Henan University of Chinese Medicine,Zhengzhou 450046,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第6期29-38,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(82174251,81573846)
北京市自然科学基金面上项目(7232300)
中国中医科学院科技创新工程项目(CI2021A00607)
中央级公益性科研院所基本科研业务费专项(YZ-202107,YZ-202148,YZX-202242)。
