Extract of Sophorae Flavescentis Radix-Cnidii Fructus couplet medicines alleviates Candida albicans-induced vulvovaginal candidiasis by activating PI3K/AKT/mTOR pathway-mediated autophagy in vaginal epithelial cells.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: The combination of Sophorae Flavescentis Radix (Ku Shen) and Cnidii Fructus (She Chuang Zi) has been widely documented in traditional Chinese medicine (TCM) formulations, demonstrating notable antibacterial, anti-inflammatory, and immunomodulatory properties. AIM OF THE STUDY: This study aims to investigate the therapeutic efficacy and mechanistic of the extract of Sophorae Flavescentis Radix-Cnidii Fructus couplet medicines (ESCC) against vulvovaginal candidiasis (VVC) induced by Candida albicans. MATERIALS AND METHODS: A murine model of VVC was established to evaluate vaginal fungal burden and inflammatory cytokine levels, while autophagy-related protein expression and RNA transcriptional profiles were systematically analyzed. A431 cells were used to assess autophagy responses after infection with C. albicans SC5314. Immunofluorescence, quantitative real-time PCR (qRT-PCR), and Western blotting (WB) were utilized to quantify autophagy-related protein expression and transcriptional activity in both murine tissues and A431 cells. Network pharmacology analysis was conducted to predict potential therapeutic targets, followed by in vitro validation of key protein expression levels in the PI3K/AKT/mTOR signaling pathway. RESULTS: In vivo, 120 mg/kg ESCC treatment restored vulvar symptoms to physiological levels in murine models, with the ESCC group exhibiting a significant reduction in vaginal fungal burden. ESCC upregulated LC3B, ATG5, and p62/SQSTM1 expression in vivo and in vitro, reversing the autophagy inhibition caused by C. albicans. Network pharmacology and KEGG enrichment analyses identified the PI3K/AKT/mTOR pathway as a pivotal mechanism, which was further verified by transcriptional and protein-level assays showing that ESCC promoted autophagic flux through pathway inhibition. CONCLUSION: ESCC alleviates C. albicans-induced VVC by activating PI3K/AKT/mTOR-mediated autophagy in vaginal epithelial cells, providing a mechanistic basis for its antifungal and supporting its potential as a plant-derived therapeutic.