Zhichi Suanzaoren Decoction alleviates perimenopausal insomnia via restoring astrocytic primary cilia and modulating Wnt/GSK-3β/GR signaling axis

Abstract

Ethnopharmacological relevance

Traditional Chinese medicine has historically used Zhichi Suanzaoren Decoction (ZSD) to alleviate perimenopausal insomnia (PMI). ZSD has demonstrated clinical efficacy in treating PMI-related symptoms; however, its active constituents and mechanisms of action remain unclear.

Aim of the study

The aim of this study was to explore the bioactive components of ZSD using UPLC-Q-TOF/MS, evaluate its efficacy in vivo experiments, and elucidate its potential mechanisms of action—particularly the role of primary hippocampal astrocyte cilia in PMI.

Materials and methods

The main components of ZSD were analyzed by UPLC-Q-TOF/MS. A PMI model was established using bilateral ovariectomy, followed by ZSD treatment. Behavioral tests and pentobarbital sodium-induced sleep synergy experiments were used to assess anti-insomnia and anxiolytic effects. Mechanistic studies included ELISA, Nissl staining, immunohistochemistry, transcriptome sequencing, immunofluorescence staining, qRT-PCR, Western blotting, and adeno-associated virus (AAV)-mediated Foxj1 knockdown.

Results

A total of 97 chemical components in the aqueous extract of ZSD were identified using UPLC-Q-TOF/MS in positive- and negative-ion modes. ZSD significantly improved sleep and reduced anxiety in PMI mice; increased hippocampal GABA levels; and reduced serum ACTH, CORT, IL-6, TNF-α, and IL-1β levels. ZSD exerts a protective effect on hippocampal neurons and reduces neuroinflammation. Transcriptomic and protein-level analyses demonstrated that ZSD inhibited Wnt signaling by promoting Foxj1 expression and restoring primary cilia in astrocytes. This led to dual effects: (1) activation of the GSK-3β/GR signaling pathway, restoring HPA axis function and reversing glucocorticoid resistance; and (2) suppression of β-catenin nuclear translocation and astrocyte-driven neuroinflammation.

Conclusions

ZSD can alleviate insomnia, anxiety, and neuroinflammation in a PMI model. The therapeutic effects are mediated through the restoration of primary astrocyte cilia via Foxj1 upregulation. These findings highlight a previously unrecognized role of primary cilia in the pathogenesis of PMI and provide a scientific rationale for the clinical application of ZSD and traditional Chinese medicine in mood-related disorders.