The Network Pharmacology of Rosmarinic Acid and Elucidating the Wound Healing Potential of a Rosmarinic Acid-Enriched Trachystemon orientalis Extract

Background/Objectives: Trachystemon orientalis (L.) D. Don has been reported to be used in folk medicine for the treatment of inflammatory wounds and for its contained rosmarinic acid, a phenolic compound known for its antioxidant, anti-inflammatory, antimicrobial, and wound healing properties. This study aimed to obtain a rosmarinic acid-enriched extract from the aerial parts of T. orientalis and evaluate its in vitro wound healing potential. Methods: Extraction conditions were optimized using fractional factorial design and Box–Behnken optimization. Rosmarinic acid content was quantified by RP-HPLC. A rosmarinic acid-enriched extract was compared with a maceration extract and isolated rosmarinic acid. Elastase, hyaluronidase, and collagenase inhibitory activities were determined. Cytotoxicity was evaluated on L929 fibroblasts using the MTT assay, and non-cytotoxic concentrations were assessed for their effects on in vitro wound healing by scratch assay. In addition, in silico target prediction and network pharmacology analyses were performed using rosmarinic acid. Results: Optimization successfully yielded a rosmarinic acid-enriched extract from T. orientalis. The rosmarinic acid-enriched extract inhibited wound-related enzymes and enhanced wound closure in the scratch assay. Network pharmacology analysis showed putative molecular targets and signaling pathways potentially associated with the wound healing activity of rosmarinic acid, especially those involved in inflammatory responses, extracellular matrix remodeling, and tissue regeneration. Conclusions: The rosmarinic acid-enriched extract exhibited promising in vitro wound healing-related activity through the inhibition of extracellular matrix degrading enzymes and promotion of wound closure. Furthermore, network pharmacology analysis provided preliminary mechanistic insights based on computational predictions.