AbstractLevels of secretory phospholipases A₂ (sPLA₂) highly increase under acute and chronic inflammatory conditions. sPLA₂ is mainly associated with high-density lipoproteins (HDL) and generates bioactive lysophospholipids implicated in acute and chronic inflammatory processes. Unexpectedly, pharmacological inhibition of sPLA₂ in patients with acute coronary syndrome was associated with an increased risk of myocardial infarction and stroke. Given that platelets are key players in thrombosis and inflammation, we hypothesized that sPLA₂-induced hydrolysis of HDL-associated phospholipids (sPLA₂-HDL) generates modified HDL particles that affect platelet function. We observed that sPLA₂-HDL potently and rapidly inhibited platelet aggregation induced by several agonists, P-selectin expression, GPIIb/IIIa activation and superoxide production, whereas native HDL showed little effects. sPLA₂-HDL suppressed the agonist-induced rise of intracellular Ca²⁺ levels and phosphorylation of Akt and ERK1/2, which trigger key steps in promoting platelet activation. Importantly, sPLA₂ in the absence of HDL showed no effects, whereas enrichment of HDL with lysophosphatidylcholines containing saturated fatty acids (the main sPLA₂ products) mimicked sPLA₂-HDL activities. Our findings suggest that sPLA₂ generates lysophosphatidylcholine-enriched HDL particles that modulate platelet function under inflammatory conditions.