基于动态代谢轨迹分析发现黄芪桂枝五物汤治疗气虚血瘀型糖尿病周围神经病变的机制

【ETHNOPHARMACOLOGICAL RELEVANCE】Huangqi Guizhi Wuwu Decoction (HGWD), a classic Traditional Chinese Medicine formula, has shown clinical efficacy in the treatment of diabetic peripheral neuropathy (DPN) with Qi deficiency and blood stasis syndrome. However, its pharmacodynamic material basis and underlying therapeutic mechanisms remain unclear.

【PURPOSE】This study aimed to systematically elucidate the pharmacodynamic material basis and therapeutic mechanisms of HGWD in the treatment of DPN using an integrated Chinmedomics strategy.

【MATERIALS AND METHODS】A rat model of DPN with Qi deficiency and blood stasis syndrome was established by streptozotocin (STZ) injection combined with dietary intervention and exhaustive swimming. HGWD was administered continuously for 8 weeks to assess its therapeutic effects. In addition to conventional pharmacological assessments, ANOVA-simultaneous component analysis (ASCA) was applied to urinary metabolomics analysis to dynamically track metabolic trajectories and characterize time-dependent changes in disease-related urinary biomarkers. Blood-absorbed components in the HGWD-treated group were characterized using UHPLC-QE-Orbitrap-MS. Correlation analysis was then performed among urinary biomarkers, key pharmacodynamic indicators, and in vivo active components. Molecular docking, molecular dynamics simulations, and in vitro experiments were further conducted to validate the core active components and their corresponding targets.

【RESULTS】HGWD significantly improved neurological function and alleviated pathological damage in the DPN rat model. Urinary metabolomics analysis identified 28 disease-related urinary biomarkers that were markedly modulated by HGWD treatment. Under conditions of confirmed therapeutic efficacy, 98 blood-absorbed components, including 63 prototype compounds and 35 metabolites, were identified. Integrated correlation analysis and network pharmacology revealed eight key pharmacodynamic components and three core targets closely associated with the therapeutic effects of HGWD. Among these components, astragaloside II, paeoniflorin, and quercetin were identified as major bioactive compounds, which may exert therapeutic effects by regulating the protein expression of P-gp, PTP1B, and MMP2.

【CONCLUSION】This study systematically clarified the pharmacodynamic material basis and therapeutic mechanisms of HGWD in the treatment of DPN using an integrated Chinmedomics approach. The integrated analysis identified 17 urinary biomarkers, 8 core components, and 3 core targets, namely ABCB1, MMP2, and PTPN1, which were further supported by molecular simulations and in vitro validation These findings provide a comprehensive strategy for deciphering the mechanisms of Traditional Chinese Medicine formulas and support the potential application of HGWD in DPN treatment.