Unrevealing the Wound Healing Involvement of TGF‐β/VEGF Signaling of Tamarix Honey‐Derived Chlorogenic Acid in ¹ H NMR Metabolomics, In Vivo, and In Silico Studies

Tamarix sp., often known as tamarisk, is a wide genus of flowering plants that reduce inflammation and cure wounds. In addition, using honey for wound therapy and care dates back thousands of years. Herein, we aimed to assess the wound healing potential of Saudi Tamarix honey (STH) metabolites in vivo, and support our hypothesis through in silico studies. Using an excision wound model, the possibility of Tamarix honey for in vivo wound healing was investigated via examining its effects on molecular targets involved in wound healing, including TGF-β, VEGF, matrix metalloproteinase-1 (MMP-1), TNF-α, and IL-1β. In addition, physicochemical properties were examined, and ¹H NMR based metabolomics technique was employed to identify STH chemical metabolites. Results revealed that Tamarix honey had a substantial impact on wound closure rate, increased levels of TGF-β and VEGF, and markedly decreased the gene expression of MMP-1, TNF-α, and IL-1β compared to the group treated with topical MEBO ointment. Biochemical and histological studies corroborated the phenotypic results. Furthermore, the effect of Tamarix honey on the redox state of wounds was investigated. On the other hand, the physical and chemical properties of STH demonstrated that it possesses moisture content 14.60%, glucose 33.7%, fructose 37.9%, sucrose 5.0%, pH 5.0, hydroxymethylfurfural (HMF) 15.40 mg/kg, and diastase enzyme 17.6 Goth scale Min, all of which that attest to its superior quality and compliance with both national and international requirements. ¹H NMR based metabolomics analysis revealed thirteen metabolites, mainly flavonoids. All Tamarix honey-derived flavonoids demonstrated high target probability against two wound healing-related targets, namely, glycogen synthase kinase-3β (GSK-3β) and NADPH oxidase 4 (NOX4). Alternatively, Tamarix honey-derived chlorogenic acid showed a high probability of targeting MMP-2, which was verified by molecular docking and molecular dynamics simulation (MDS). Our study highlights the potential of Tamarix honey in wound repair by revealing the most likely signaling pathway utilizing in silico and gene expression analysis.