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.