Baihe Ganmai Dazao Decoction ameliorates depression-like and sleep-disturbance phenotypes: potential involvement of IL-17A/IL-17RA-related inflammatory signaling and microglial polarization.

ETHNOPHARMACOLOGICAL RELEVANCE: Originally documented in Zhang Zhongjing's Synopsis of Prescriptions of the Golden Chamber of the Eastern Han Dynasty, Ganmai Dazao Decoction (GMDZ) is composed of three medicinal materials: Glycyrrhizae Radix et Rhizoma (GRR), Tritici Levis Fructus (TLF), and Jujubae Fructus (JF). Traditionally, it has been used to treat zangzao, a syndrome characterized by emotional disturbance and sleep disorder, and is now widely prescribed for depression, anxiety, and insomnia. Nevertheless, the effective material foundation and intrinsic therapeutic mechanisms of GMDZ are still not thoroughly clarified. Based on GMDZ, Baihe Ganmai Dazao Decoction (BGMD) was developed by adding Lilii Bulbus (LB), Poria (PO), and Ziziphi Spinosae Semen (ZSS) to strengthen its Yin-nourishing and mind-calming effects. AIM OF THE STUDY: This study aimed to optimize the compatibility ratio of BGMD, elucidate its pharmacodynamic material basis and antidepressant and sleep-promoting mechanisms, and establish a quality control strategy. MATERIALS AND METHODS: The optimal compatibility ratio of BGMD was screened using a uniform design combined with the entropy weight method in zebrafish models of depression and sleep disturbance. To further characterize its therapeutic activities, a composite animal model coupling chronic unpredictable mild stress (CUMS) stimulation and 4-chloro-DL-phenylalanine (PCPA) administration was constructed to mimic depressive states alongside sleep dysfunction in mice. UPLC-Q-TOF-MS/MS, network pharmacology, bioinformatics, machine learning, and molecular docking were integrated to screen putative active constituents and key targets, followed by validation using RT-qPCR, immunohistochemistry, ELISA, and immunofluorescence. HPLC-based quantification of quality markers was performed, and the in vivo activities of candidate constituents were further verified in zebrafish. RESULTS: The optimal compatibility ratio of BGMD was TLF: GRR: JF: LB: PO: ZSS = 5: 3: 1: 6: 4: 2. BGMD effectively ameliorated depression-like behaviors and sleep dysfunction in model mice. It markedly upregulated the hippocampal contents of gamma-aminobutyric acid (GABA), dopamine (DA), 5-hydroxytryptamine (5-HT) and norepinephrine (NE), while reducing the levels of interleukin-1β (IL-1β) and corticosterone (CORT). BGMD corrected the imbalance in microglial M1/M2 phenotypes and alleviated neuronal damage within the CA1, CA3, and DG subregions of the hippocampus. Mechanistically, BGMD may exert its effects partly through modulation of IL-17A/IL-17RA-related inflammatory signaling and associated downstream signaling molecules, accompanied by suppression of Il17a, Il17ra, Traf6, p-STAT3, and p-JNK3. Eighty-six compounds were identified in BGMD, and eight of them were selected as candidate active constituents. Regaloside A, spinosin, and liquiritin were selected as representative quality-control indicator compounds, with mean contents of 0.0500, 0.0364, and 0.0844 mg/g, respectively. CONCLUSION: BGMD exerts synergistic antidepressant and sleep-promoting effects, which may be associated with modulation of IL-17A/IL-17RA-related inflammatory signaling, restoration of microglial polarization balance, protection of hippocampal neuronal integrity, and improvement of neurotransmitter homeostasis. These findings provide insights into the pharmacodynamic basis and potential mechanisms of BGMD and support its further quality-control standardization and pharmacological investigation.