Using a combination of in vitro and in silico approaches to explore the biological potential of Arceuthobium oxycedri extracts: innovative insights from the lab to functional applications

In the present study, the antioxidant capacity, enzyme inhibitory activities, and chemical composition of various extracts from Arceuthobium oxycedri were assessed by LC-MS/MS. Extracts were further evaluated for their cytotoxicity by the resazurin reduction assay and for their antioxidant activity by qRT-PCR in liver carcinoma HepG2 cells. The LC-MS/MS indicated that in both ethanolic and ethanol/water (70%) extracts, there were highly identified phenolic compounds, and they exhibited greater antioxidant effects. These extracts exhibited potential inhibitory activities against acetylcholinesterase and butyrylcholinesterase with the values of 4.72 and 4.41 mg GALAE/g, respectively. The ethanol extract showed the highest α-glucosidase inhibitory activity, 2.19 mmol ACAE/g, probably due to its high content of proanthocyanidins and hydroxycinnamic acids. Furthermore, results demonstrate that some of the extracts significantly reduce the expression of the antioxidant gene HO-1, while γ-GCS was slightly increased. To further investigate the molecular mechanisms, network pharmacology approach was used to identify potential targets related to HO-1 and NRF2.The analysis revealed key proteins associated with oxidative stress and metabolic pathways. Subsequently, molecular docking was performed on these identified targets, along with standard enzymes such as AChE, BChE, α-amylase, αglucosidase, and tyrosinase. The best-performing protein-ligand interactions were then selected for molecular dynamics simulations. These findings enable us to infer that ethanol and ethanol/water (70%) extracts represent the most promising candidates based on their chemical composition for their medicinal importance, represented by their rich contents of phenolic and flavonoid compounds.