摘要
目的:探讨温阳解郁方调节“母婴分离+束缚应激(MS+RS)”抑郁小鼠海马神经元凋亡改善突触可塑性的机制。方法:仔鼠出生第0天(PD0)随机分为空白组10只和造模组50只。造模采用MS+RS建立抑郁模型,PD21离乳后随机分为模型组、温阳组、解郁组、温阳解郁组、氟西汀组,每组10只,PD21~PD111分组给药。糖水偏好、开放旷场、O迷宫及新物体识别行为实验评估小鼠焦虑抑郁和学习记忆能力;免疫组化法(IHC)检测小鼠海马突触后致密物95(PSD95)表达情况;原位末端标记法(TUNEL)染色检测小鼠海马神经元凋亡情况;蛋白免疫印迹法(Western blot)检测海马脑源性神经营养因子(BDNF)、磷酸化酪氨酸蛋白激酶/酪氨酸蛋白激酶(p-TrkB/TrkB)、磷酸化蛋白激酶B/蛋白激酶B(p-Akt/Akt)、磷酸化哺乳动物雷帕霉素靶蛋白/哺乳动物雷帕霉素靶蛋白(p-mTOR/mTOR)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶-3(Caspase-3)、突触后致密蛋白95(PSD95)、突触素(Syn)蛋白表达情况。结果:与空白组比较,模型组小鼠糖水偏好程度、中央区停留时间、运动总距离、开臂停留时间和认知指数显著减少(P<0.01),IHC结果显示海马PSD95表达明显减少,海马神经元凋亡数量增加(P<0.01),BDNF、p-TrkB/TrkB、p-Akt/Akt、p-mTOR/mTOR、Bcl-2、p-mTOR/mTOR、PSD95、Syn蛋白表达显著减少(P<0.01),Bax、Caspase-3蛋白含量明显增加(P<0.05);与模型组比较,温阳解郁组和氟西汀组糖水偏好程度、5 min中央运动时间、5 min开臂停留时间和认知指数明显增加(P<0.05,P<0.01),海马PSD95表达明显增加、海马凋亡指数显著增加(P<0.01),BDNF、p-TrkB/TrkB、p-Akt/Akt、p-mTOR/mTOR、Bcl-2、PSD95、Syn蛋白含量显著增加(P<0.01),Bax、Caspase-3蛋白含量显著减少(P<0.01)。结论:温阳解郁方可以通过BDNF/Akt/mTOR信号通路,改善模型组小鼠神经元凋亡情况,保护海马突触结构和功能,缓解模型组小鼠抑郁样�
Objective:To explore the mechanism of Wenyang Jieyu prescription in regulating hippocampal neuron apoptosis and improving synaptic plasticity in the mouse model of depression induced by maternal separation combined with restraint stress.Method:The mice on postnatal day 0(PD0)were randomly assigned into a control group(n=10)and a modeling group(n=50).Maternal separation combined with restraint stress was adopted to establish the mouse model of depression,and the modeled mice were randomized into model,Wenyang prescription,Jieyu prescription,Wenyang Jieyu prescription,and fluoxetine groups(n=10)on the weaning day(PD21).From PD21 to PD111,the mice were fed with the diets mixed with corresponding medicines.The sucrose preference test,open field test,O-maze test,and novel object recognition test were then conducted to evaluate the depression,memory,and learning abilities of mice.Immunohistochemistry(IHC)was employed to measure the atomic absorbance(AA)of postsynaptic density protein 95(PSD95)in the hippocampus.Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling(TUNEL)was employed to detect the apoptosis of hippocampal neurons.Western blot was employed to determine the protein levels of brainderived neurotrophic factor(BDNF),phosphorylated tyrosine kinase receptor B/tyrosine kinase receptor B(p-TrkB/TrkB),phosphorylated protein kinase B/protein kinase B(p-Akt/Akt),phosphorylated mammalian target of rapamycin/mammalian target of rapamycin(p-mTOR/mTOR),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X(Bax),cysteinyl aspartate-specific proteinase-3(Caspase-3),synaptophysin(Syn),and PSD95.Result:Compared with the control group,the modeling decreased the sucrose preference rate,time spent in central zone within 5 min,total movement distance,time spent in the open arm,and cognition index(P<0.01).Furthermore,it decreased the expression of PSD95,increased the neuron apoptosis in the hippocampus(P<0.01),down-regulated the protein levels of BDNF,p-TrkB/TrkB,p-Akt/Akt,p-mTOR/mTOR,Bcl-2,PSD95,and Syn(P<0.01),and up-regulat
作者
孟丹华
佘楷杰
孟晓莹
巩子汉
梁文青
王英
梁媛
岳广欣
MENG Danhua;SHE Kaijie;MENG Xiaoying;GONG Zihan;LIANG Wenqing;WANG Ying;LIANG Yuan;YUE Guangxin(Institute of Basic Theroy for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Basic Medicine,Henan University of Chinese Medicine,Zhengzhou 450000,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第6期48-57,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)
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-202153,YZ-202216)。
