Anmei Dan Regulates Hippocampal Inflammation via CX3CL1/CX3CR1 Signaling Pathway to Improve Learning and Memory in Aged Sleep-deprived Mice
10.13422/j.cnki.syfjx.20260303
- VernacularTitle:安寐丹调控CX3CL1/CX3CR1信号通路介导海马炎症改善老年睡眠剥夺小鼠学习记忆的作用机制
- Author:
Lichun WANG
1
;
Zi'ao WANG
1
;
Jinxu MA
1
;
Yufeng CAI
2
;
Huizhen LIU
2
;
Ping WANG
2
;
Guangjing XIE
2
Author Information
1. School of Basic Medical Sciences， Hubei University of Chinese Medicine，Wuhan 430065，China
2. Engineering Research Center of Chinese Medicine Protection Technology and New Product Development for Elderly Brain Health of Ministry of Education，Hubei University of Chinese Medicine， Wuhan 430065，China
- Publication Type:Journal Article
- Keywords:
Anmei Dan;
aged sleep deprivation;
neuroinflammation;
learning and memory;
C-X3-C motif chemokine ligand 1 （CX3CL1）/C-X3-C motif chemokine receptor 1 （CX3CR1） signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2026;32(17):97-106
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effects of Anmei Dan （AMD） on the expression of proteins in the C-X3-C motif chemokine ligand 1 （CX3CL1）/C-X3-C chemokine receptor 1 （CX3CR1） signaling pathway， hippocampal inflammation， and learning and memory in an aged sleep-deprived model. MethodsSixty aged C57 mice were randomly assigned into a control group， a model group， a melatonin group （1.3 mg·kg-1·d-1）， and high-， medium-， and low-dose AMD groups （26.26， 13.13， 6.565 g·kg-1·d-1， respectively）， with 10 mice per group. Continuous sleep deprivation was administered for 4 weeks through a custom-built sleep deprivation chamber. The cognitive function of mice was assessed via the Y-maze test. Histomorphological alterations in pyramidal cells of the hippocampal CA1 and DG regions were examined by hematoxylin-eosin （HE） and Nissl staining. Synaptic morphology in the hippocampus was visualized with Golgi staining. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of inflammatory cytokines interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the hippocampal tissue as well as the level of interleukin-10 （IL-10） in the prefrontal cortex. Real-time PCR was performed to detect markers of M1-type ［cluster of differentiation （CD） 86 and inducible nitric oxide synthase （iNOS）］ and M2-type ［CD206 and arginase 1 （Arg1）］ macrophages. Western blot was employed to quantify the protein levels of CX3CL1， CX3CR1， phosphorylated nuclear factor κB p65 subunit （p-NF-κB p65）， and phosphorylated p38 mitogen-activated protein kinase （p-p38 MAPK） in the hippocampus. Immunofluorescence double staining was performed to co-label CX3CR1 with ionized calcium-binding adapter molecule 1 （Iba-1） for observation of the co-localization of target proteins with microglia. ResultsCompared with the control group， the model group exhibited disrupted daily activity patterns with reduced spontaneous alternation rates （P<0.01）， disorganized morphology and arrangement of hippocampal neurons， accompanied by reduced numbers of Nissl bodies and dendritic spines （P<0.01）， upregulated protein levels of IL-6， IL-10， IL-1β， TNF-α， CX3CL1， CX3CR1， p-NF-κB p65， p-p38 MAPK， and Iba-1 and mRNA levels of CD86 and iNOS （P<0.01）， and downregulated mRNA levels of CD206 and Arg1 （P<0.05，P<0.01）. Compared with the model group， the melatonin group and medium- and high-dose AMD groups showed increased spontaneous alternation rates （P<0.01）， improved neuronal morphology， number， and spine density in the hippocampal CA1 and DG regions （P<0.01）， declined levels of IL-6， IL-10， IL-1β， and TNF-α （P<0.05， P<0.01）， downregulated protein levels of CX3CL1， CX3CR1， p-NF-κB p65， p-p38 MAPK， and Iba-1 and mRNA levels of CD86 and iNOS， and upregulated mRNA levels of CD206 and Arg1 （P<0.01）. ConclusionAMD may improve the learning and memory in aged sleep-deprived mice by mediating neuroinflammation through the CX3CL1/CX3CR1 signaling pathway.
