Multi-target healing mechanisms of Hippobroma longiflora for diabetic wounds: A comprehensive network pharmacology study, molecular docking, and experimental insights

Abstract

Objective: This research investigated the multi-target mechanisms of Hippobroma longiflora (HL) in promoting diabetic wound healing using an integrated approach involving network pharmacology, molecular docking, and both in vitro and in vivo evaluations. Materials and Methods: The bioactive compounds of HL leaves' 70% ethanol extract were detected using liquid chromatography-high-resolution mass spectrometry (LC-HRMS), and potential targets were predicted via the SwissTargetPrediction. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analyses were conducted using DAVID. Molecular docking was predicted using MOE version 2015.10. In vitro assays using HaCaT cells evaluated proliferation and Tumor Necrosis Factor-alpha (TNF-α) levels via ELISA. In vivo measurement in diabetic rats induced by streptozotocin (n = 9 per group), with histopathological analysis conducted on days 7, 14, and 21. Results: HL extract exhibited anti-inflammatory and pro-proliferative effects, primarily mediated by syringic acid (P41) and vanillactic acid (P43). These compounds upregulated AKT1, MAPK3, and HIF-1α. HL treatment significantly enhanced HaCaT cell proliferation (p < 0.0001) and reduced TNF-α level at 31.25 ppm (p < 0.05). In vivo, HL treatment decreased inflammatory cell infiltration from day 7, with significant tissue regeneration observed by day 14 (p < 0.01) relative to base control. Conclusion: HL and its bioactive constituents demonstrate promising potential for diabetic wound healing by modulating inflammatory mediators (TNF-α) and enhancing key regenerative pathways (AKT1, MAPK3, and HIF-1α).