Fenugreek ( Trigonella foenum‐graecum L.) as a functional food associated with improved heat‐stress survival in Caenorhabditis elegans

Abstract BACKGROUND Fenugreek ( Trigonella foenum‐graecum L.) is widely consumed as a functional food across Mediterranean, Middle Eastern, and South Asian cultures and has long been valued in traditional diets for its health‐promoting properties. Although fenugreek contains diverse phytochemicals with known biological activities, its potential effects on organismal resistance to heat stress remain insufficiently characterized. RESULTS In this study, the effects of fenugreek seed preparations on thermotolerance‐associated survival were evaluated using Caenorhabditis elegans . Two complementary extraction approaches were investigated: an aqueous decoction (TW) and a hydroethanolic macerate (TE). Synchronized adult worms were exposed to the extracts before heat stress at 35 °C, and survival was analyzed using the Kaplan–Meier method and restricted mean survival time (RMST). Both extracts were associated with significantly improved heat‐stress survival compared to the control group ( P < 0.001). The RMST values were 73.91 min for TW, 62.55 min for TE and 38.44 min for the control group; however, the difference between the two extract preparations was not statistically significant. MTT [i.e. 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide] assays in CCD‐1079Sk human dermal fibroblasts showed concentration‐dependent reductions in metabolic activity, with 24‐h half‐maximal inhibitory concentration values of 1.85 μg mL −1 for TE and 12.63 μg mL −1 for TW. Liquid chromatography–high‐resolution mass spectrometry profiling identified several constituents, with orientin being the most abundant quantified compound in both extracts. Molecular docking of selected fenugreek‐associated compounds against human AKT serine/threonine kinase 1; protein kinase B alpha and C. elegans serine/threonine‐protein kinase AKT‐1 revealed favorable predicted binding affinities. Subsequent molecular dynamics simulations suggested that orientin remained associated with both targets under the simulated conditions, although temperature‐dependent differences were observed in the dynamics of the C. elegans AKT‐1 complex. CONCLUSION These findings indicate that fenugreek seed preparations are associated with improved heat‐stress survival in C. elegans . The numerically higher RMST observed with the decoction, despite its lower orientin content, suggests that the observed biological activity may involve the combined effects of multiple constituents; however, the difference between the two extract preparations was not statistically significant. These findings support further dose–response and mechanistic studies to clarify the association between fenugreek preparations and improved heat‐stress survival. © 2026 Society of Chemical Industry.