Exploring Effect of Method of Warming Yang and Relieving Depression on Susceptibility to Mental Disorders in a Two-hit Mice via BDNF/TrkB Signaling Pathway
10.13422/j.cnki.syfjx.20260340
- VernacularTitle:从BDNF/TrkB信号通路探讨温阳解郁法对二次打击小鼠精神障碍易感性的影响
- Author:
Zihan GONG
1
;
Jingwen YANG
2
;
Ying WANG
3
;
Wenqing LIANG
4
;
Guangxin YUE
5
Author Information
1. The Ningxia Hui Autonomous Region Chinese Medicine Hospital and Chinese Medicine Research Institute， Yinchuan 750021， China
2. Institute of Basic Research in Clinical Medicine， China Academy of Chinese Medical Sciences， Beijing 100700， China
3. Xiyuan Hospital of China Academy of Chinese Medical Sciences， Beijing 100091， China
4. Hengshanqiao People's Hospital， Changzhou Economic Development Zone， Changzhou 213119， China
5. Institute of Basic Theory for Chinese Medicine， China Academy of Chinese Medical Sciences， Beijing 100700， China
- Publication Type:Journal Article
- Keywords:
warming Yang and relieving depression;
two-hit;
anxiety- and depression-like behaviors;
brain-derived neurotrophic factor （BDNF）/tyrosine kinase receptor B （TrkB） signaling pathway;
mice
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2026;32(19):154-163
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the mechanism underlying the effects of three formulas of the method of warming Yang and relieving depression on anxiety- and depression-like behaviors in the mice subjected to a two-hit paradigm of maternal separation （MS） combined with restraint stress （RS）. MethodsNeonatal mice were randomly assigned into the following groups： Blank control， MS alone， model （MS+RS）， warming Yang （WY，5.85 g·kg-1）， relieving depression （RD，12.03 g·kg-1）， warming Yang and relieving depression （WYRD，16.71 g·kg-1）， and fluoxetine groups （2.6 mg·kg-1）. MS was conducted on postnatal day 5 （PD5）. Weaning and preventive drug administration were carried out on PD21， and restraint stress was applied on PD90. Anxiety- and depression-like behaviors were assessed using the open field test （OFT）， the elevated zero maze test and tail suspension test （TST）. High-performance liquid chromatography with electrochemical detection （HPLC-ECD） was employed to measure the levels of neurotransmitters， including 5-hydroxytryptamine （5-HT） and dopamine （DA）， in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression levels of 5-HT1A receptor （5-HT1AR）， serotonin transporter （SERT）， monoamine oxidase A （MAOA）， glucocorticoid receptor （GR）， adrenocorticotropic hormone （ACTH）， corticotropin-releasing hormone （CRH）， brain-derived neurotrophic factor （BDNF）， and tyrosine kinase receptor B （TrkB）. Immunohistochemistry （IHC） was applied to assess the protein expression levels of 5-HT1AR， 5-HT2A receptor （5-HT2AR）， 5-HT2C receptor （5-HT2CR）， GR， and CRH receptor 1 （CRHR1）. The protein expression levels of BDNF and TrkB in the hippocampus were measured using the fully automated protein expression analysis system （Simple Wes）. ResultsIn the behavioral tests， compared with the blank control group， the model group exhibited significant increases in locomotor distance and central area crossings （P<0.05， P<0.01）in OFT， as well as a significant increase in immobility time in the TST （P<0.05）. Compared with the model group， the RD， WYRD， and fluoxetine groups all showed significantly reduced central area crossings （P<0.05， P<0.01）. Regarding neurotransmitter levels， compared with the blank control group， the model group had significantly decreased hippocampal epinephrine （E） and dopamine （DA） content （P<0.05， P<0.01）. Compared with the MS group， the model group showed significant decreases in hippocampal norepinephrine （NE）， E， and 5-HT content （P<0.05， P<0.01）. Compared with the model group， the WY group decreased hippocampal NE， DA， and 5-HT content （P<0.01）， while the RD， WYRD， and fluoxetine groups exhibited significantly increased hippocampal E content （P<0.01）. At the molecular level， compared with the blank control and MS groups， the model group showed significant increases in mRNA levels of 5-HT1AR， SERT， MAOA， ACTH， and CRH （P<0.01）， and significant decreases in mRNA levels of tryptophan hydroxylase （TPH）， GR， BDNF， and TrkB （P<0.01）. Protein expression of 5-HT1AR， 5-HT2AR， and 5-HT2CR was also significantly elevated （P<0.05， P<0.01） in the model group. Compared with the model group， the RD group showed significant decreases in SERT， MAOA， and CRH mRNA （P<0.05， P<0.01）， significant increases in TPH， BDNF， and TrkB mRNA （P<0.01）， significantly reduced protein expression of 5-HT1AR， 5-HT2AR， and 5-HT2CR （P<0.05）， and significantly increased GR protein expression. The WY group showed a significant increase in ACTH mRNA expression （P<0.01）. The fluoxetine group exhibited significant decreases in MAOA and CRH mRNA （P<0.05， P<0.01）， significant increases in TPH， BDNF， and TrkB mRNA （P<0.01）， significantly reduced 5-HT2AR protein expression （P<0.01）， and significantly increased GR protein expression（P<0.05）. ConclusionThe method of warming Yang and relieving depression can ameliorate anxiety- and depression-like behaviors in two-hit mice. Its mechanism may be associated with the BDNF/TrkB signaling pathway， as well as the HPA axis and serotonergic system.
