Lysophosphatidylcholine induces oxidative stress and calcium‐mediated cell death in human blood platelets

Abstract

AbstractPlatelets are essential component of circulation that plays a major role in hemostasis and thrombosis. During activation and its demise, platelets release platelet‐derived microvesicles, with lysophosphatidylcholine (LPC) being a prominent component in their lipid composition. LPC, an oxidized low‐density lipoprotein, is involved in cellular metabolism, but its higher level is implicated in pathologies like atherosclerosis, diabetes, and inflammatory disorders. Despite this, its impact on platelet function remains relatively unexplored. To address this, we studied LPC's effects on washed human platelets. A multimode plate reader was employed to measure reactive oxygen species and intracellular calcium using H2DCF‐DA and Fluo‐4‐AM, respectively. Flow cytometry was utilized to measure phosphatidylserine expression, mitochondrial membrane potential (ΔΨm), and mitochondrial permeability transition pore (mPTP) formation using FITC‐Annexin V, JC‐1, and CoCl2/calcein‐AM, respectively. Additionally, platelet morphology and its ultrastructure were observed via phase contrast and electron microscopy. Sonoclot and light transmission aggregometry were employed to examine fibrin formation and platelet aggregation, respectively. The findings demonstrate that LPC induced oxidative stress and increased intracellular calcium in platelets, resulting in increased phosphatidylserine expression and reduced ΔΨm. LPC triggered caspase‐independent platelet death and mPTP opening via cytosolic and mitochondrial calcium, along with microvesiculation and reduced platelet counts. LPC increased the platelet's size, adopting a balloon‐shaped morphology, causing membrane fragmentation and releasing its cellular contents, while inducing a pro‐coagulant phenotype with increased fibrin formation and reduced integrin αIIbβ3 activation. Conclusively, this study reveals LPC‐induced oxidative stress and calcium‐mediated platelet death, necrotic in nature with pro‐coagulant properties, potentially impacting inflammation and repair mechanisms during vascular injury.