Ethyl acetate extract of Sargentodoxa cuneata and Patrinia villosa alleviates pelvic inflammation by shifting macrophages polarization.

BACKGROUND: Pelvic inflammatory disease (PID) is a prevalent gynecological infection, that poses significant therapeutic challenges. Sargentodoxa cuneata and Patrinia villosa (S&P) demonstrate empirical efficacy in PID treatment within traditional Chinese medicine practice. However, their active components and precise anti-inflammatory mechanisms should be systematically elucidated.

PURPOSE: This study aimed to investigate the pharmacologically active components of S&P and decipher their anti-inflammatory mechanism through integrated multi-omics approaches.

MATERIALS AND METHODS: CCK-8 and NO inhibition assays were used to screen the optimal extract from three solvent partitions (ethyl acetate [EtOAc], n-butanol [n-BuOH], and water). The chemical components of different extracts and S&P were detected by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The effects of EtOAc extract on PID were studied in an lipopolysaccharide (LPS) -induced PID murine model to evaluate uterine histopathology and macrophage phenotype. RNA sequencing was utilized to analyze the differentially expressed genes after EtOAc extract treatment. Cytokines and macrophage phenotype markers were detected by flow cytometry. LC-MS/MS-based metabolic analysis and western blotting were used to study the potential mechanism.

RESULTS: Bioactivity screening identified EtOAc extract as the most potent fraction, suppressing iNOS-mediated NO production without cytotoxicity. EtOAc extract inhibited the release of IL-6 and upregulated the M2 phenotypic marker CD206 in LPS induced RAW264.7 and THP-1 cells. In the murine PID model, EtOAc extract treatment alleviated LPS-induced pelvic inflammation, reduced uterine F4/80+ and CD86+ (M1) macrophages, and upregulated CD206+ (M2) subset. Transcriptomics of uterine tissue demonstrated that EtOAc extract upregulated genes related to M2 macrophages polarization, tissue repair and vascular remodeling, and endometrial receptivity and lactic acid metabolism, and downregulated genes involved in M1 macrophages, inflammation, and glycolysis and lactic acid synthesis. Metabolomics analysis showed that EtOAc extract ameliorated LPS-induced metabolic disorder. The differentially expressed metabolites were involved in the tricarboxylic acid (TCA) cycle, arginine and proline metabolism, pyruvate metabolism, and glycolysis or gluconeogenesis pathways. EtOAc extract inhibited the expression of glucose metabolism key proteins HKI, HKII, PKM2 and PD, and NF-κB and IκB-ɑ phosphorylation, and synergized with HK-II inhibitor (2-deoxy-D-glucose).

CONCLUSIONS: EtOAc extract alleviates LPS-induced PID via a novel immunometabolic axis, namely, by suppressing HK-II-mediated glycolysis through the NF-κB axis and balancing macrophages polarization. The mechanistic elucidation of macrophage immunometabolic reprogramming by S&P provides a molecular basis for phytotherapy in PID management.