The Chinese ancient prescription Zhentonggufang alleviates pain and inflammation, which is associated with reduced levels of inflammatory cytokines IL-6 and IL-1β

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Chronic pain poses serious health and socioeconomic burdens, highlighting the urgent need for safer, more effective analgesic alternatives from natural sources. This study aimed to explore the analgesic and anti-inflammatory potential and mechanisms of Zhentonggufang (ZTGF), a traditional Chinese prescription, and to develop a more effective drug delivery system in the form of a microemulsion hydrogel to enhance its clinical applicability. We integrated data related to the discovery and development of analgesic and anti-Ninflammatory agents to construct an online web-based discovery platform. From this platform, ZTGF was identified as a candidate traditional Chinese ancient prescription for analgesia. For the first time, its mechanism of action was investigated using network pharmacology and molecular docking. The analgesic efficacy and mechanism of ZTGF were further validated through acetic acid-induced writhing, formalin-induced pain, and carrageenan-induced paw edema tests in rats, along with enzyme-linked immunosorbent assay (ELISA) analysis. A microemulsion hydrogel formulation of ZTGF was then prepared and characterized using Fourier-transform infrared spectroscopy (FTIR), viscosity analysis, particle size distribution, and zeta potential measurement. The enhanced efficacy of the formulation was further confirmed in vivo. Two online platforms were established: the Analgesia Ancient Prescription Database (AAPD), containing herbal extracts and traditional formulas, and the Analgesia Factor Database (AFD), containing isolated natural products and synthetic compounds. Network pharmacology analysis revealed quercetin, ursolic acid, and other compounds as key active ingredients of ZTGF, which act on targets including IL-6, TNF-α, and IL-1β, primarily through the PI3K-AKT and MAPK signaling pathways. Pharmacological evaluations demonstrated that ZTGF significantly attenuated pain and inflammatory responses in animal models. ELISA results further revealed that ZTGF administration markedly decreased the expression of the pro-inflammatory cytokines IL-6 and IL-1β. In addition, physicochemical analyses indicated that the formulated hydrogel possessed favorable stability and effectively mitigated inflammation in Complete Freund's Adjuvant (CFA)-induced rats. ZTGF alleviated pain and inflammation in animal models, which may be associated with reduced levels of IL-6 and IL-1β. The developed hydrogel showed good physicochemical stability and provided an effective topical delivery system for ZTGF.

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