Exploring the Underlying Mechanism of Action of <i>Lippia nodiflora</i> on Wound Healing Based on Network Pharmacology, Molecular Docking, and Experimental Validation

Lippia nodiflora leaves have been traditionally used by healers in Silent Valley, Palakkad district of Kerala, for treating wounds. This study aimed to scientifically validate the wound-healing potential of the methanol extract of L. nodiflora leaves (MELN) to support its ethnopharmacological claims. Phytochemical profiling using UPLC-QTOF-MS identified key constituents, and network pharmacology analysis revealed potential pathways and targets involved in wound healing. These findings were further supported by molecular docking, in vitro, in vivo, and reverse transcription quantitative polymerase chain reaction (RT-qPCR) analyses. No toxicity was observed in either in vitro or in vivo models. Network pharmacology indicated that MELN may act through the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor resistance pathway. Core compounds such as quercetin, 3-methylherbacetin, eupafolin, 6-hydroxyluteolin 3'-methyl ether 7-sulfate, and syringic acid beta-glucopyranosyl ester likely contribute to the wound-healing activity, as supported by docking studies. RT-qPCR analysis demonstrated upregulation of genes related to cell migration and proliferation. In vitro assays showed enhanced cell proliferation and migration, indicating pro-healing properties. In vivo excision wound models confirmed significant wound closure, complete epithelialization, and improved histopathological features in MELN-treated groups. Overall, this study provides scientific evidence supporting the ethnopharmacological claim of L. nodiflora in wound management and establishes MELN as a potential herbal formulation for wound healing.