Bakh formula ameliorates high-fat diet-induced erectile dysfunction in rats via coordinated restoration of metabolic and tissue homeostasis.

Introduction: Erectile dysfunction (ED) associated with long-term high-fat diet (HFD) intake is increasingly recognized as a manifestation of systemic metabolic and vascular dysfunction. This study aimed to evaluate the therapeutic effects of the classical Uyghur medicine formula Bakh Formula (BAKHF) on HFD-induced ED in rats and elucidate its underlying mechanisms. Methods: Male rats with HFD-induced ED were randomly assigned to model, Yimusake (YMSK, positive control), and BAKHF groups. YMSK served as a pharmacological benchmark to validate the model and evaluate BAKHF's effects on erectile function, histology, and hematology. Erectile function was evaluated by maximal intracavernosal pressure to mean arterial pressure (ICP/MAP), sexual behavior tests, and biochemical assays. The primary outcome was erectile function assessed by ICP/MAP, with secondary outcomes including metabolic, endocrine, molecular, and histopathological parameters. The potential mechanisms of BAKHF were elucidated using integrated transcriptomic, proteomic, and network pharmacology analyses, combined with chemical profiling of serum-absorbed compounds, molecular docking, RT-qPCR, and Western blot. Results: Both BAKHF and YMSK improved erectile function, characterized by elevated ICP/MAP ratios and improved sexual behavior parameters. Histologically, both formulas restored penile cavernous structure, reduced collagen deposition, and increased smooth muscle content, confirming YMSK's role as a reliable phenotypic benchmark for the model. However, BAKHF exhibited superior efficacy in restoring systemic metabolic and endocrine homeostasis, including normalization of food intake and serum lipid profiles. Subsequent mechanistic analyses focused on BAKHF and identified its regulatory role in AMPK-associated metabolic and cytoskeletal pathways. Five serum-absorbed compounds, including rosmarinic acid and ferulic acid, were identified as potential bioactive constituents targeting key nodes like GAPDH, PPARγ, and ESR1. Discussion: These findings suggest that BAKHF acts through multi-component, multi-target, and multi-pathway synergy, potentially involving AMPK signaling to regulate metabolic and tissue homeostasis in HFD-induced ED. The integration of multi-omics analyses with experimentally confirmed serum-absorbed compounds and mechanistic validation strengthens the biological plausibility of the findings. However, limitations include the lack of pathway-specific genetic or pharmacological interventions and unresolved contributions of individual constituents. Targeting systemic metabolic and tissue homeostasis through multi-component herbal formulas may represent a complementary therapeutic strategy for metabolism-related ED.