Enhanced Antiplatelet Activity of Nitrated Fatty Acid Extracts from Phaseolus vulgaris L

Dietary bioactive compounds are increasingly explored as complementary cardioprotective strategies, and the nitration of unsaturated fatty acids has emerged as a process capable of enhancing antiplatelet properties. This study investigated whether Phaseolus vulgaris L. extracts can generate nitrated fatty acids under gastric-like conditions and evaluated their effects on human platelet function. Bean extracts and major fatty acids were nitrated in vitro and tested using washed platelets to assess cytotoxicity, TRAP-6 and collagen-induced aggregation, activation markers (P-selectin, CD63), and mitochondrial responses including membrane potential, ROS production, and Ca²⁺ dynamics. Nitrated extracts markedly inhibited TRAP-6 induced aggregation (IC₅₀ ≈ 1.8 mg/mL), whereas non-nitrated extracts showed minimal activity; this effect was reversed by β-mercaptoethanol, indicating dependence on electrophilic nitroalkenes. Fractionation revealed that the lipidic fraction accounted for most of the antiplatelet effect, and isolated nitrated fatty acids (NO₂-LN, NO₂-LA, NO₂-OA) displayed stronger inhibition than their native counterparts without increasing cytotoxicity. Nitrated species additionally reduced mitochondrial membrane potential and granule secretion without elevating ROS. These findings identify Phaseolus vulgaris L. as a natural source of bioactive nitrated fatty acids and support their potential as nutraceutical agents capable of modulating platelet activation and contributing to cardiovascular risk reduction.