Abstract

This study conducted a comprehensive in vitro pharmacological evaluation of the crude extract and solvent-partitioned fractions of Gnetum gnemon var. tenerum. LC-MS metabolite profiling identified 77 compounds, including flavonoids and phenylethanolamine derivatives, that exhibit the antioxidant, cytotoxic, and anti-inflammatory activities described in this paper. The total phenolic and flavonoid contents (TPC/TFC) were quantified, showed that the ethyl acetate and n-butanol fractions contained the highest levels of these phytochemicals. These fractions exhibited strong antioxidant activity in both ABTS and DPPH assays. The antibacterial activity of the extracts was assessed using disk diffusion, MIC, and MBC assays against five pathogenic bacteria, where the chloroform and ethyl acetate fractions demonstrated the highest potency, with MIC values as low as 0.049 mg/mL against Acinetobacter baumannii. Cytotoxicity testing on human gastrointestinal cancer cell lines (AGS, HT-29) and a normal intestinal epithelial cell line (HIEC-6) showed dose-dependent cytotoxicity toward cancer cells while exerting markedly lower effects on normal cells, suggesting selective action. Anti-inflammatory activity, evaluated by nitric oxide (NO) inhibition in LPS-stimulated RAW264.7 macrophages, revealed that the chloroform and ethyl acetate fractions most effectively suppressed NO production. These findings provide scientific support for the traditional uses of G. gnemon and highlight its extracts as a promising source of bioactive compounds with multi-target pharmacological potential against oxidative stress, bacterial infections, inflammation, and cancer.