Changes of Microglia in Hippocampus of Mice Induced by Maternal Separation with Restraint Stress and Regulatory Effect of Wenyang Jieyu Prescription

Kai-jie SHE; Jing-jing GAO; Zi-han GONG; Huan-run ZHANG; Yang ZUO; Jing-wen YANG; Guang-xin YUE; Yuan LIANG

doi:10.13422/j.cnki.syfjx.20211837

您当前的位置：

首页 >

文章列表页 >

Changes of Microglia in Hippocampus of Mice Induced by Maternal Separation with Restraint Stress and Regulatory Effect of Wenyang Jieyu Prescription

Pharmacology | 更新时间：2021-08-18

- Changes of Microglia in Hippocampus of Mice Induced by Maternal Separation with Restraint Stress and Regulatory Effect of Wenyang Jieyu Prescription
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 18, Pages: 49-57(2021)
- 作者机构：
  
  1.中国中医科学院中医基础理论研究所，北京 100700
  2.开封市中医院，河南开封 475000
  3.北京中医药大学第三附属医院，北京 100029
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20211837
  CLC： R2-0;R749.7+2;R33
- Received：12 May 2021，
  
  Published Online：27 July 2021，
  
  Published：20 September 2021
- 稿件说明：
移动端阅览
佘楷杰,高静静,巩子汉等.母婴分离/束缚应激模型小鼠海马小胶质细胞变化及温阳解郁方的调节作用[J].中国实验方剂学杂志,2021,27(18):49-57.

SHE Kai-jie,GAO Jing-jing,GONG Zi-han,et al.Changes of Microglia in Hippocampus of Mice Induced by Maternal Separation with Restraint Stress and Regulatory Effect of Wenyang Jieyu Prescription[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(18):49-57.
佘楷杰,高静静,巩子汉等.母婴分离/束缚应激模型小鼠海马小胶质细胞变化及温阳解郁方的调节作用[J].中国实验方剂学杂志,2021,27(18):49-57. DOI： 10.13422/j.cnki.syfjx.20211837.

SHE Kai-jie,GAO Jing-jing,GONG Zi-han,et al.Changes of Microglia in Hippocampus of Mice Induced by Maternal Separation with Restraint Stress and Regulatory Effect of Wenyang Jieyu Prescription[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(18):49-57. DOI： 10.13422/j.cnki.syfjx.20211837.

摘要

目的

观察母婴分离/束缚应激诱发焦虑抑郁小鼠海马小胶质细胞激活和白细胞介素-1

（IL-1

），白细胞介素-6（IL-6），肿瘤坏死因子-

（TNF-

）表达情况，探讨温阳解郁方抗焦虑抑郁的机制。

方法

84只雄性C57BL仔鼠于出生后第0天（PD0）随机分为空白组、束缚应激组和造模组，造模组采取母婴分离结合束缚应激制备焦虑抑郁模型，将造模组按随机数字表法分为模型组、温阳组、解郁组、温阳解郁组和氟西汀组。在离乳第21天（PD21）至成年第90天（PD90），空白组、束缚应激组和模型组饲喂正常饲料，其余饲喂药混饲料（温阳组、解郁组、温阳解郁组和氟西汀组用药剂量分别为5.85，12.03，16.71 g·kg

-1

及2.60 mg·kg

-1

）。采用旷场实验，O迷宫和社会交互实验评估模型组小鼠焦虑抑郁状态；免疫组化法（IHC）观察海马小胶质细胞及离子钙接头蛋白-1（Iba-1）表达；实时荧光定量聚合酶链式反应（Real-time PCR）检测IL-1

，IL-6，TNF-

，Iba-1，糖皮质激素受体（GR）mRNA表达水平。

结果

与空白组比较，模型组小鼠5 min运动总路程和中央区活动时间显著减少（

0.01），开臂活动时间与活动总路程明显减少（

0.05，

0.01），训练探察时间和辨别探查时间显著增加（

0.01），Iba-1蛋白及mRNA表达，IL-1

β，

IL-6，TNF-

mRNA表达水平显著升高（

0.01），GR mRNA表达水平显著降低（

0.01），IHC结果显示小胶质细胞的过度激活。与模型组比较，温阳解郁方及氟西汀组小鼠5 min运动总路程和中央区活动时间显著上升（

0.01），开臂活动时间显著上升（

0.01），训练探察时间和辨别探查时间明显降低（

0.05，

0.01）；Iba-1蛋白及mRNA表达，IL-1

，IL-6，TNF-

mRNA表达水平明显降低（

0.05，

0.01），GR mRNA表达水平显著上升（

0.01），IHC结果表明小胶质细胞的形态恢复；温阳组和解郁组小鼠开臂活动时间显著上升（

0.01），辨别探查时间明显降低（

0.05），Iba-1蛋白及mRNA表达，IL-6 mRNA表达水平明显降低（

0.05，

0.01）；温阳组小鼠GR mRNA表达水平明显上升（

0.05），TNF-

mRNA表达水平明显降低（

0.05）；解郁组小鼠5 min运动总路程显著上升（

0.01），训练探察时间明显降低（

0.05），IL-1

mRNA表达水平明显降低（

0.05），IHC结果表明两组小胶质细胞有所恢复。

结论

温阳解郁方的综合疗效和药理作用优于温阳方和解郁方，其作用机制可能通过抑制母婴分离/束缚应激小鼠海马小胶质细胞的激活，降低IL-1

，IL-6，TNF-

等细胞因子的表达，增加海马GR表达，从而改善小鼠的焦虑抑郁样行为。

Abstract

Objective

To observe the activation of microglia in hippocampus of depressed and anxious mice induced by maternal separation with acute restraint stress and the expression of interleukin-1

（IL-1

），interleukin-6（IL-6），tumor necrosis factor-

（TNF-

）， investigating the mechanism of Wenyang Jieyu prescription in treating anxiety and depression.

Method

Eighty four male C57BL offspring were randomly divided into control group， acute restraint stress group and model group on postnatal day 0（PD0）. Maternal separation combined with acute restraint stress was used to prepare anxious and depressed model mice， dividing the model mice into model group， Wenyang， Jieyu， Wenyang Jieyu and fluoxetine group according to random number table method. During the period of PD21-PD90， the control， acute restraint stress and model mice were fed with normal diet， with the other groups fed with corresponding medicine mixed diet. The Wenyang， Jieyu and Wenyang Jieyu groups were given 5.85， 12.03 and 16.71 g·kg

-1

·d

-1

respectively. The fluoxetine group was given 2.60 mg·kg

-1

·d

-1

. Open field， zero maze test and social interaction tests were used to evaluate the anxiety and depression of model mice. The expression of Iba-1 in hippocampal microglia was detected by immunohistochemistry（IHC）. The mRNA expression of IL-1

， IL-6， TNF-

， Iba-1 and glucocorticoid receptor（GR）were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）.

Result

Compared with the control group， total movement distance and time spent in central zone in 5 min of the model mice significantly decreased（

0.01）， time spent in opened arm and total movement distance decreased significantly（

0.05，

0.01）， investigation time during testing and training increased significantly（

0.01）. The expression of Iba-1 protein and mRNA，IL-1

，IL-6，TNF-

mRNA significantly increased（

0.01）， the expression levels of GR mRNA significantly decreased（

0.01）. The result of IHC staining showed that microglia were over activated. Compared with the model group， total movement distance and time spent in central zone in 5 min of mice in the Wenyang Jieyu and fluoxetine group significantly increased（

0.05，

0.01）.Time spent in opened arm significantly increased（

0.01）. Investigation time during testing and training significantly decreased（

0.05，

0.01）. The expression of Iba-1 protein and mRNA，IL-1

，IL-6，TNF-

mRNA significantly decreased（

0.05，

0.01）. The expression of GR mRNA increased significantly（

0.05，

0.01）. IHC staining showed the microglia recovered. Time spent in opened arm of mice in the Wenyang group and Jieyu group significantly increased （

0.01）， time spent investigating during testing decreased significantly （

0.05）， the expression levels of Iba-1 protein and mRNA，IL-6 mRNA significantly decreased （

0.05，

0.01）. The expression of GR mRNA of mice in the Wenyang group significantly increased （

0.05）， the expression of TNF-

mRNA significantly decreased （

0.05）. Total movement distance of mice in the Jieyu group increased significantly （

0.01）， time spent investigating during training decreased significantly （

0.05），the expression level of IL-1

mRNA significantly decreased （

0.05）. IHC staining showed that microglia recovered partly in both groups.

Conclusion

The comprehensive curative effect and pharmacological action of Wenyang Jieyu prescription were better than Wenyang prescription and Jieyu prescription. Wenyang Jieyu prescription can treat anxiety and depression in maternal separation and acute restraint stress mice， its possible mechanism may be related to the decreased activation of microglia， down-regulation of IL-1

，IL-6，TNF-

expression and up-regulation of GR expression.

关键词

Keywords

references

李传朋，刘玉，魏品球，等 . 逍遥散及其类方与有效成分抗抑郁作用机制研究进展［J］. 中国实验方剂学杂志， 2020 ， 26 （ 6 ）： 243 - 250 .

MOKDAD A H ， FOROUZANFAR M H ， DAOUD F ， et al . Global burden of diseases，injuries，and risk factors for young people's health during 1990-2013：a systematic analysis for the Global Burden of Disease Study 2013 ［J］. Lancet ， 2016 ， 387 （ 10036 ）： 2383 - 2401 .

崔利军，栗克清，严保平，等 . 抑郁症共病其他精神障碍的特点及相关因素［J］. 中国心理卫生杂志， 2010 ， 24 （ 8 ）： 592 - 596，603 .

韩岳 . 小胶质细胞在早期生活应激所致行为异常中的作用机制研究［D］：成都：电子科技大学， 2020 .

BANQUERI M ， MÉNDEZ M ， GÓMEZ-LÁZARO E ， et al . Early life stress by repeated maternal separation induces long-term neuroinflammatory response in glial cells of male rats ［J］. Stress ， 2019 ， 22 （ 5 ）： 563 - 570 .

GEHRAND A L ， HOEYNCK B ， JABLONSKI M ， et al . Programming of the adult HPA axis after neonatal separation and environmental stress in male and female rats ［J］. Endocrinology ， 2018 ， 159 （ 7 ）： 2777 - 2789 .

HARTMANN J ， BAJAJ T ， KLENGEL C ， et al . Mineralocorticoid receptors dampen glucocorticoid receptor sensitivity to stress via regulation of FKBP5 ［J］. Cell Rep ， 2021 ， 35 （ 9 ）： 109185 .

岳广欣，张玲，卢贺起，等 . 温阳解郁法对母婴分离小鼠行为模式及HPA轴功能的影响［J］. 中国中医基础医学杂志， 2014 ， 20 （ 1 ）： 42 - 46 .

巫鑫辉，李娜，岳广欣 . 留守儿童身心健康问题与中医理论认识［J］. 世界中西医结合杂志， 2016 ， 11 （ 8 ）： 1170 - 1172 .

刘燕，邹小娟，丁秀芳，等 . 基于复合应激的肝郁脾虚证小鼠模型的建立与评价［J］. 中华中医药杂志， 2016 ， 31 （ 5 ）： 1840 - 1844 .

吴丹，高耀，邢婕，等 . 逍遥散治疗肝郁脾虚型抑郁症的药理作用机制研究进展［J］. 中国实验方剂学杂志， 2019 ， 25 （ 8 ）： 187 - 193 .

罗武龙，陈洁，牛婕，等 . 基于网络药理学的二仙汤治疗抑郁症的作用机制研究［J］. 中国药理学通报， 2020 ， 36 （ 9 ）： 1317 - 1324 .

梁媛，张玲，卢贺起，等 . 温阳解郁不同组方对母婴分离/社会击败应激小鼠影响的比较研究［J］. 时珍国医国药， 2014 ， 25 （ 9 ）： 2274 - 2277 .

王小雪，岳广欣，巫鑫辉，等 . 抑郁症海马神经可塑性改变及中药调控作用研究现状［J］. 辽宁中医药大学学报， 2017 ， 19 （ 4 ）： 80 - 84 .

VAN ZYL P J ， DIMATELIS J J ， RUSSELL V A . Behavioural and biochemical changes in maternally separated Sprague-Dawley rats exposed to restraint stress ［J］. Metab Brain Dis ， 2016 ， 31 （ 1 ）： 121 - 133 .

FRANKLIN T B ， LINDER N ， RUSSIG H ， et al . Influence of early stress on social abilities and serotonergic functions across generations in mice ［J］. PLoS One ， 2011 ， 6 （ 7 ）： e21842 .

陈世洲，毛国庆，孙玉明，等 . 加味二仙汤治疗阳虚质骨质疏松症临床疗效及机制［J］. 中国实验方剂学杂志， 2020 ， 26 （ 7 ）： 104 - 108 .

纪雅菲，芮翊馨，方洋，等 . 逍遥散正丁醇部位基于IGF-1R β /PI3K/Akt信号通路的抗抑郁作用［J］. 中国实验方剂学杂志， 2021 ， 27 （ 14 ）： 1 - 11 .

VETULANI J . Early maternal separation：a rodent model of depression and a prevailing human condition ［J］. Pharmacol Rep ， 2013 ， 65 （ 6 ）： 1451 - 1461 .

KULKARNI S K ， SINGH K ， BISHNOI M . Elevated zero maze：a paradigm to evaluate antianxiety effects of drugs ［J］. Methods Find Exp Clin Pharmacol ， 2007 ， 29 （ 5 ）： 343 - 348 .

GREGUS A ， WINTINK A J ， DAVIS A C ， et al . Effect of repeated corticosterone injections and restraint stress on anxiety and depression-like behavior in male rats ［J］. Behav Brain Res ， 2005 ， 156 （ 1 ）： 105 - 114 .

刘玲 . 内侧前额叶皮层抑制性神经元在社会交互行为调控中的作用及神经机制研究［D］：杭州：浙江大学， 2020 .

MATROV D ， KõIV K ， KANARIK M ， et al . Middle-range exploratory activity in adult rats suggests higher resilience to chronic social defeat ［J］. Acta Neuropsychiatr ， 2016 ， 28 （ 3 ）： 125 - 140 .

RESHETNIKOV V ， RYABUSHKINA Y ， KOVNER A ， et al . Repeated and single maternal separation specifically alter microglial morphology in the prefrontal cortex and neurogenesis in the hippocampus of 15-day-old male mice ［J］. Neuroreport ， 2020 ， 31 （ 18 ）： 1256 - 1264 .

WANG Y L ， HAN Q Q ， GONG W Q ， et al . Microglial activation mediates chronic mild stress-induced depressive- and anxiety-like behavior in adult rats ［J］. J Neuroinflammation ， 2018 ， 15 （ 1 ）： 21 .

CALCIA M A ， BONSALL D R ， BLOOMFIELD P S ， et al . Stress and neuroinflammation：a systematic review of the effects of stress on microglia and the implications for mental illness ［J］. Psychopharmacology （Berl）， 2016 ， 233 （ 9 ）： 1637 - 1650 .

ROSENBLAT J D ， MCINTYRE R S ， ALVES G S ， et al . Beyond monoamines-novel targets for treatment-resistant depression：a comprehensive review ［J］. Curr Neuropharmacol ， 2015 ， 13 （ 5 ）： 636 - 655 .

MILLER A H ， RAISON C L . The role of inflammation in depression：from evolutionary imperative to modern treatment target ［J］. Nat Rev Immunol ， 2016 ， 16 （ 1 ）： 22 - 34 .

KRISHNAN V ， NESTLER E J . The molecular neurobiology of depression ［J］. Nature ， 2008 ， 455 （ 7215 ）： 894 - 902 .

BEUREL E ， TOUPS M ， NEMEROFF C B . The bidirectional relationship of depression and inflammation：double trouble ［J］. Neuron ， 2020 ， 107 （ 2 ）： 234 - 256 .

LEONARD B E . Inflammation and depression：a causal or coincidental link to the pathophysiology？［J］. Acta Neuropsychiatr ， 2018 ， 30 （ 1 ）： 1 - 16 .

THIBAUT F . Neuroinflammation：new vistas for neuropsychiatric research ［J］. Dialogues Clin Neurosci ， 2017 ， 19 （ 1 ）： 3 - 4 .

HAN F ， OZAWA H ， MATSUDA K ， et al . Colocalization of mineralocorticoid receptor and glucocorticoid receptor in the hippocampus and hypothalamus ［J］. Neurosci Res ， 2005 ， 51 （ 4 ）： 371 - 381 .

WILBER A A ， SOUTHWOOD C J ， WELLMAN C L . Brief neonatal maternal separation alters extinction of conditioned fear and corticolimbic glucocorticoid and NMDA receptor expression in adult rats ［J］. Dev Neurobiol ， 2009 ， 69 （ 2/3 ）： 73 - 87 .

ANACKER C ， CATTANEO A ， MUSAELYAN K ， et al . Role for the kinase SGK1 in stress，depression，and glucocorticoid effects on hippocampal neurogenesis ［J］. Proc Natl Acad Sci U S A ， 2013 ， 110 （ 21 ）： 8708 - 8713 .

DE BOSSCHER K ， VANDEN BERGHE W ， HAEGEMAN G . Mechanisms of anti-inflammatory action and of immunosuppression by glucocorticoids：negative interference of activated glucocorticoid receptor with transcription factors ［J］. J Neuroimmunol ， 2000 ， 109 （ 1 ）： 16 - 22 .

MANIAM J ， MORRIS M J . Voluntary exercise and palatable high-fat diet both improve behavioural profile and stress responses in male rats exposed to early life stress：role of hippocampus ［J］. Psychoneuroendocrinology ， 2010 ， 35 （ 10 ）： 1553 - 1564 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Mechanism of Wenyang Jieyu Prescription in Regulating Activation of Mouse Hippocampal Microglia Based on JAK2/STAT3 Signaling Pathway

Effect of Curcumin on Cognitive Function in Mouse Model of Low Oxygen-induced Chronic Nerve Injury

Effect of Xiaoyaosan on Apoptosis in Mouse Model of Lipopolysaccharide-induced Depressive-like Behavior in Mice

Medication Ideas for Depression from Wenyang Jieyu Prescription

Wenyang Jieyu Prescription Regulates Hippocampal Neuron Apoptosis and Improves Synaptic Plasticity in Depressed Mice via BDNF/Akt/mTOR Pathway

Related Author

WANG Ying

GONG Zihan

LIANG Wenqing

YANG Jingwen

YUE Guangxin

LI Gaifen

LI Jiaxin

SUN Zhibo

Related Institution

Ningxia Chinese Medicine Research Center

Hengshanqiao People's Hospital in Changzhou Economic Development Zone

Institute of Basic Research in Clinical Medicine，China Academy of Chinese Medical Sciences

Institute of Basic Theory for Traditional Chinese Medicine，China Academy of Chinese Medical Sciences

Institute of Basic Theory of Traditional Chinese Medicine（TCM），China Academy of Chinese Medical Sciences

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰