| Summary: | <i>Glycyrrhiza glabra</i> and <i>Sophora japonica</i> (Fabaceae) are well-known medicinal plants with valuable secondary metabolites and pharmacological properties. The flavonoid-rich fractions of <i>G. glabra</i> roots and <i>S. japonica</i> leaves were prepared using Diaion column chromatography, and the confirmation of flavonoid richness was confirmed using UPLC-ESI-MS profiling and total phenolics and flavonoids assays. UPLC-ESI-MS profiling of the flavonoid-rich fraction of <i>G. glabra</i> roots and <i>S. japonica</i> leaves resulted in the tentative identification of 32 and 23 compounds, respectively. Additionally, the wound healing potential of topical preparations of each fraction, individually and in combination (1:1) ointment and gel preparations, were investigated in vivo, supported by histopathological examinations and biomarker evaluations, as well as molecular docking studies for the major constituents. The topical application of <i>G. glabra</i> ointment and gel, <i>S. japonica</i> ointment and gel and combination preparations significantly increase the wound healing rate and the reduction of oxidative stress in the wound area via MDA reduction and the elevation of reduced GSH and SOD levels as compared to the wound and Nolaver<sup>®</sup>-treated groups. The molecular docking study revealed that that major compounds in <i>G. glabra</i> and <i>S. japonica</i> can efficiently bind to the active sites of three proteins related to wound healing: glycogen synthase kinase 3-<i>β</i> (GSK3-<i>β</i>), matrix metalloproteinases-8 (MMP-8) and nitric oxide synthase (iNOS). Consequently, <i>G. glabra</i> roots and <i>S. japonica</i> leaves may be a rich source of bioactive metabolites with antioxidant, anti-inflammatory and wound healing properties.
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