Jiedu-Shengji Ointment promotes diabetic wound repair by inhibiting the NLRP3/Caspase-1/GSDMD signaling pathway and regulating macrophage function.

INTRODUCTION: Diabetic ulcers are one of the most common clinical complications of diabetes. These ulcers are characterized by poor healing and high recurrence rates, which lead to heavy medical burdens and greatly elevate the risk of limb amputation. They have therefore become a critical challenge for clinical tissue repair. Jiedu-Shengji Ointment (JDSJG) is a topical preparation composed of scorpio, scolopendra, frankincense and myrrh. Existing studies have verified its efficacy in promoting diabetic wound healing. Nevertheless, the specific molecular mechanisms responsible for its wound-healing activity remain poorly elucidated. This study aimed to preliminarily explore the molecular mechanisms by which JDSJG facilitates diabetic wound repair.

METHODS: We established a mouse model of diabetic skin wounds using streptozotocin (STZ) induction combined with full-thickness skin puncture. We then detected and evaluated multiple indicators of wound tissue, including tissue microstructure and repair status, skin macrophage polarization, the mRNA and protein levels of inflammatory factors, as well as the key gene and protein expression of the NLRP3/Caspase-1/GSDMD signaling pathway. In the in vitro experiment, we induced pyroptosis in RAW264.7 cells via lipopolysaccharide (LPS) stimulation. We further observed cellular morphological changes, detected cell pyroptosis and macrophage polarization, and measured the expression of inflammatory factors and key molecules in the NLRP3/Caspase-1/GSDMD pathway. Finally, we adopted LC-MS/MS and GO and KEGG enrichment analyses to clarify the material basis and potential molecular mechanisms of JDSJG.

RESULTS AND DISCUSSION: In a mouse model of diabetic wounds, treatment with JDSJG effectively alleviated the inflammatory response and significantly promoted the healing process of diabetic wounds in mice. In vitro and in vivo experiments confirmed that JDSJG downregulated the expression of inflammatory cytokines and inhibited the activation of the NLRP3/Caspase-1/GSDMD pathway. Additionally, flow cytometry results indicated that JDSJG can induce the polarization of macrophages toward the anti-inflammatory M2 phenotype. LC-MS/MS and network pharmacology analyses confirmed that the primary ingredients of JDSJG-scorpio and scolopendra-are associated with pyroptosis. Overall, this study established the therapeutic efficacy of JDSJG in promoting diabetic wound repair and highlighted its potential in regulating wound inflammation and macrophage polarization.