Huiyang Shengji Unguent Promotes Lymphangiogenesis and Wound Healing in Diabetic Chronic Wounds: Combined Insights from Proteomics and in vivo and in vitro Analyses.

Purpose: To investigate the mechanism by which HYSJ unguent promotes lymphangiogenesis and improves the healing of diabetic chronic wounds (DCWs). Methods: The main components of HYSJ were identified by mass spectrometry. A mouse model with chronic skin ulcers was established. The ultrastructure and lymphatic drainage of lymphatic endothelial cells in wounds were examined. Lymphatic markers in wound tissues were detected, and proteomic and bioinformatics analyses were performed to identify differentially expressed proteins and associated pathways. In vitro, a high-glucose inflammatory environment that mimics DCWs was induced in human lymphatic endothelial cells (HLEC). HYSJ and caspase inhibitors were used for intervention. Diverse assays were conducted to assess HLEC function and activation of inflammatory cell death. Results: The primary constituents of HYSJ unguent included coclaurine, sinapine, and ononin, among others. HYSJ increased healing of DCWs in diabetic mice, protected lymphatic endothelial cells, restored lymphatic drainage in the wound, and upregulated expression of key lymphatic proteins. Numerous proteins, including TLR2, Myd88, STAT1, and inflammatory response-related proteins such as NLRP3, Caspase-1, GSDMD, were expressed differentially in mice. HYSJ unguent also stimulated expression of key lymphatic proteins in HLEC, protected cell function, and suppressed inflammatory cell death. Conclusion: HYSJ unguent enhances lymphangiogenesis, protects lymphatic endothelial cells in a high-glucose inflammatory environment, and accelerates DCW healing by suppression of TLR2/Myd88/caspase-1 signaling pathway. These findings provide important experimental support for the pharmacological mechanism by which HYSJ unguent facilitates healing of DCWs.