A Anti-Angiogenic and Anti-Proliferative Effects of Juglans regia Green Husk Extract: Experimental and In Silico Evaluation: Juglans regia Green Husk extract

Angiogenesis is a regulated process essential for tissue repair, but it also plays a role in cancer development through vascular endothelial growth factor (VEGF)-mediated pathways. Natural products, including the green husk of Juglans regia L., are considered promising anti-angiogenic compounds. This study aimed to elucidate the anti-angiogenic potential of Juglans regia L., green husk ethanolic extract through experimental, phytochemical, and computational analyses. An angiogenesis assay was used to assess the potential to interfere with blood vessel formation. The extract's impact on endothelial cell growth was investigated using the MTT test in the HUVEC cell model. VEGF (vascular endothelial growth factor) expression was measured using the process of reverse transcription polymerase chain reaction (RT-qPCR). The phytochemical characterization of the extract was analyzed through high-performance liquid chromatography (HPLC). Molecular docking, SwissADME pharmacokinetic prediction, in silico toxicity, and metabolic biotransformation prediction analyses were also conducted. The ethanolic extract showed concentration-dependent inhibition of microvessel sprouting and HUVEC proliferation, with IC₅₀ values of 37 µg/mL and 49.2 µg/mL, respectively. RT-qPCR analysis demonstrated downregulation of VEGF gene expression to 0.702-fold relative to the untreated control. HPLC analysis identified rutin as a major flavonoid constituent of the extract. Molecular docking revealed favorable interactions of rutin aglycone with VEGFR2, while computational analyses indicated acceptable pharmacokinetic, metabolic, and toxicity profiles. The extract demonstrated promising anti-angiogenic and anti-proliferative activities and may represent a potential natural VEGFR2-targeting agent.