Integration of network pharmacology and experimental verification to reveal the active constituents and molecular mechanism of Compound Fujin Shengji (FJSJ) Powder in the treatment of diabetes ulcer

Objective Fujin Shengji (FJSJ) Powder has a positive effect on healing Diabetic Ulcers (DU), but the specific compounds at work have not been identified. Methods Network pharmacology was employed to investigate the primary constituents, potential therapeutic targets, and pharmacological mechanisms of FJSJ Powder treating DU. In vitro experiments were further performed to validate these findings: an LPS-induced RAW264.7 cell model was established to assess the anti-inflammatory effects of FJSJ Powder active constituents treating DU, while a high glucose-induced HUVEC model was cultured to verify the pro angiogenic effect of FJSJ Powder active constituents treating DU. Additionally, the mechanisms by which the optimal active monomers suppress inflammation and enhance angiogenesis in DU were investigated, focusing on the JAK2/STAT3 and PI3K/AKT signaling pathways. Results Network pharmacology identified quercetin (Que), kaempferol (Kae), abietic acid (Aba) and acetyl-11-keto-β-boswellic acid (AKBA) as the primary active constituents of FJSJ Powder treating DU; IL6, TNF, AKT1, and VEGFA were the core targets; and anti-inflammatory effect was mainly enriched in the JAK2/STAT3 signaling pathway, while angiogenesis promotion was mainly enriched in the PI3K/AKT signaling pathway. The vitro experiments have confirmed Que, Kae, Aba, and AKBA could play important roles in anti-inflammatory and angiogenic effects, with AKBA and Aba playing important roles in anti-inflammatory and pro-angiogenic effects, respectively. AKBA can upregulate P-JAK2, P-STAT3, IL-4, and downregulate INOS protein levels to exert anti-inflammatory. Aba can upregulate the P-AKT, P-PI3K, HIF-1α, and VEGFA proteins levels to promote angiogenesis. Conclusion FJSJ Powder promotes DU healing through dual-pathway regulation in anti-inflammatory and pro-angiogenic effects, with AKBA mediating the best anti-inflammatory activity through the JAK2/STAT3 pathway and Aba driving the strongest pro-angiogenic effects via the PI3K/AKT/HIF-1α/VEGFA axis, providing a basis for formula optimization.