Asian Journal of Food Research and Nutrition

Introduction: Plant-derived functional foods are increasingly recognized for their role in managing different health-related risk factors. Despite the known traditional uses of fenugreek seed, its potential and influence on nitric oxide production require further scientific validation.

Aim: To investigate the role of Standardized fenugreek seed extract (Testosurge® and FLG) in enhancing eNOS phosphorylation and nitric oxide production to support endothelial function under in vitro conditions.

Study Design: In vitro experimental study using a Human Umbilical Vein Endothelial Cell (HUVEC) model to evaluate dose-dependent enzymatic signaling and bioactive efficacy.

Study Location: BioTeSys GmbH, Schelzorstrasse 54-56, 73728 Esslingen, Germany, from November 2023 to January 2024.

Methodology: HUVECs were treated with varying concentrations of Standardized fenugreek seed extract. Extracts underwent simulated gastrointestinal digestion prior to treatment of HUVECs. The activity of eNOS was quantified using an indirect immunofluorescence assay specifically targeting phosphorylation at the Serine (S1176) residue. Subsequent enzymatic activation was measured to correlate nitric oxide production with functional output. Significances were calculated using Student’s t-test or One-way ANOVA and Dunnett’s post test.

Results: The extract demonstrated a clear dose-dependent increase in eNOS phosphorylation at S1176. This upregulation may lead to a significant increase in NO production compared to the control groups. The data confirms that the phytochemicals within the extract (flavonoids, alkaloids, and saponins) directly stimulate the eNOS signaling pathway. Testosurge® resulted in a 117% activation at 5 minutes compared to vehicle control.

Conclusion: Standardized fenugreek seed extracts may support endothelial function which leads to improved vascular tone and blood flow. Further in vivo and clinical studies are required to confirm these findings.