Shengji Yuhong Ointment accelerates diabetic wound healing by suppressing AGEs/RAGE-related inflammation and restoring HIF-1α/VEGF-mediated angiogenesis.

ETHNOPHARMACOLOGICAL RELEVANCE: Shengji Yuhong Ointment (SJYHO) is a classic external formula in traditional Chinese medicine surgery, which was first recorded in Wai Ke Zheng Zong. It is commonly used for conditions such as chronic non-healing sores and ulcers, failure of necrotic tissue to slough off, poor granulation tissue formation, and chronic refractory wounds. Diabetic wounds are long exposed to a pathological microenvironment characterized by hyperglycemia, chronic inflammation, oxidative stress, and impaired angiogenesis. This shares intrinsic connections with the traditional indications of SJYHO. However, the modern pharmacological basis and key mechanisms of action of SJYHO for diabetic wound repair have not been fully elucidated. In particular, it is still unclear whether the wound healing effect of intact SJYHO involves the synergistic regulation of AGEs/RAGE-related inflammation, macrophage polarization, and angiogenesis. AIM OF THE STUDY: This study aims to systematically evaluate the intervention effect of SJYHO on diabetic wound healing, and explore its potential mechanism from the aspects of AGEs/RAGE-related inflammatory response, oxidative stress, macrophage polarization, and HIF-1α/VEGF-mediated angiogenesis, with pharmacological validation using the RAGE inhibitor (FPS-ZM1). MATERIALS AND METHODS: This study used UHPLC-MS/MS to characterize the main chemical components of SJYHO and combined network pharmacology to predict its potential targets and related pathways. Diabetic wounds were established in STZ-induced diabetic mice, and treated topically with Vaseline or SJYHO for 10 consecutive days. Furthermore, FPS-ZM1, a selective RAGE inhibitor, was used for pharmacological validation of RAGE involvement. Wound area measurement, H&E staining, Masson staining, immunohistochemistry, immunofluorescence, ELISA, biochemical marker assays, RT-qPCR, and Western blot were conducted to analyze wound closure, tissue repair, collagen deposition, angiogenesis, inflammatory response, oxidative stress, and macrophage polarization status. RESULTS: UHPLC-MS/MS analysis identified that SJYHO contains Ferulic acid, Shikonin, Loureirin, and other representative ingredients. Network pharmacology analysis showed that its potential targets are mainly enriched in the AGE-RAGE signaling pathway in diabetic complications, HIF-1 signaling pathway, VEGF signaling pathway, TNF signaling pathway, and inflammation-related pathways. Animal experiments showed that SJYHO significantly promoted wound closure, reducing the residual wound area on Day 10 from 36.37 ± 5.10% in the DM group to 24.47 ± 3.01% in the DM + SJYHO group (P < 0.05), while increasing granulation tissue thickness and collagen deposition and enhancing CD31-positive angiogenesis. Mechanistically, SJYHO reduced the expression of AGEs and RAGE in wound tissues, inhibited the elevation of ROS and MDA, restored SOD activity, and downregulated the levels of TNF-α, IL-1β, IL-6, and p-p65/p65. At the same time, SJYHO partially decreased the expression of the M1-type marker iNOS and upregulated the expression of the M2-type markers ARG1 and CD206. Additionally, SJYHO partially restored the expression of HIF-1α and VEGF in wound tissues and promoted HIF-1α/CD31-related angiogenic responses. FPS-ZM1 intervention further supported AGEs/RAGE signaling in the process of improving the diabetic wound microenvironment and promoting tissue repair in SJYHO. CONCLUSIONS: SJYHO could effectively promote diabetic wound healing, and its effects are closely related to inhibiting AGEs/RAGE-related oxidative stress and inflammatory response, regulating macrophage M1/M2 polarization, restoring HIF-1α/VEGF-mediated angiogenesis and improving collagen remodeling. This study provides a reference basis for SJYHO as a classic Chinese medicine external formula for the treatment of chronic refractory diabetic wounds.