Integrated evaluation of Reseda lutea extracts through biological assays, zebrafish toxicity, and network pharmacology

Reseda lutea is a rich source of bioactive phytochemicals, whose composition and biological properties may vary substantially depending on the extraction conditions. In this study, aerial parts of Reseda lutea were extracted using solvents of different polarities, including ethyl acetate (EA), ethanol, ethanol/water, and water, and their chemical composition and biological activities were comparatively evaluated. Total phenolic and flavonoid contents were determined by Folin–Ciocalteu and AlCl3 assays, while phytochemical profiling was performed by HPLC-MS/MS. The extracts exhibited distinct phytochemical compositions depending on the extraction solvent. Antioxidant capacity was assessed using DPPH, ABTS, CUPRAC, and FRAP assays, with the ethanol/water and ethanol extracts showing the strongest activities. Enzyme inhibitory effects were evaluated against AChE, BChE, tyrosinase, α-amylase, and α-glucosidase. Ethanol and EA extracts demonstrated the highest anti-AChE and anti- BChE activities, whereas the ethanol/water extract showed the strongest anti-tyrosinase effect. Cytotoxicity analyses revealed dose-dependent anticancer activity of the water extract against HELA and DU-145 cell lines. In addition, zebrafish assays revealed concentration-dependent embryotoxicity of the water extract, characterized by increased mortality, delayed hatching, and edema-related morphological abnormalities during early embryogenesis, whereas post-hatching larvae showed markedly lower sensitivity. Overall, R. lutea extracts exhibited solvent-dependent chemical and biological properties, with ethanol and ethanol/water extracts showing the most pronounced bioactivities. These findings highlight the therapeutic potential of R. lutea while emphasizing the importance of developmental toxicity assessment for its safe application.