Lysophosphatidic acid induces thrombogenic activity through phosphatidylserine exposure and procoagulant microvesicle generation in human erythrocytes

College of Pharmacy, Seoul National University, Seoul 151-742, Korea.
Arteriosclerosis Thrombosis and Vascular Biology (Impact Factor: 6). 03/2007; 27(2):414-21. DOI: 10.1161/01.ATV.0000252898.48084.6a
Source: PubMed


Although erythrocytes have been suggested to play a role in blood clotting, mediated through phosphatidylserine (PS) exposure and/or PS-bearing microvesicle generation, an endogenous substance that triggers the membrane alterations leading to a procoagulant activity in erythrocytes has not been reported. We now demonstrated that lysophosphatidic acid (LPA), an important lipid mediator in various pathophysiological processes, induces PS exposure and procoagulant microvesicle generation in erythrocytes, which represent a biological significance resulting in induction of thrombogenic activity.
In human erythrocytes, LPA treatment resulted in PS exposure on remnant cells and PS-bearing microvesicle generation in a concentration-dependent manner. Consistent with the microvesicle generation, scanning electron microscopic study revealed that LPA treatment induced surface changes, alteration of normal discocytic shape into echinocytes followed by spherocytes. Surprisingly, chelation of intracellular calcium did not affect LPA-induced PS exposure and microvesicle generation. On the other hand, protein kinase C (PKC) inhibitors significantly reduced PS exposure and microvesicle generation induced by LPA, reflecting the role of calcium-independent PKC. Activation of PKC was confirmed by Western blot analysis showing translocation of calcium-independent isoform, PKCzeta, to erythrocyte membrane. The activity of flippase, which is important in the maintenance of membrane asymmetry, was also inhibited by LPA. Furthermore, LPA-exposed erythrocytes actually potentiated the thrombin generation as determined by prothrombinase assay and accelerated the coagulation process initiated by recombinant human tissue factor in plasma. The adherence of erythrocytes to endothelial cells, another important feature of thrombogenic process, was also stimulated by LPA treatment.
These results suggested that LPA-exposed erythrocytes could make an important contribution to thrombosis mediated through PS exposure and procoagulant microvesicle generation.

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    • "The role of the erythrocytes in the pathogenesis of thrombosis has been not well studied. In recent years a number of reports have found that they have an important role in initiating and sustaining a procoagulant response by acting as a source of phospholipids or triggering the coagulation system through the activation of a number of coagulation factors [17] [18] [19]. Furthermore, Kozuma and coworkers demonstrated that PNH erythrocytes-derived microparticles have important procoagulant properties [20]. "
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    • "At least in theory, phosphatidylserine exposing erythrocytes may further interfere with microcirculation [72,73,74,75,76,77], as phosphatidylserine exposing erythrocytes adhere to endothelial CXCL16/SR-PSO [73], stimulate blood clotting and trigger thrombosis [72,78,79]. Baicalein has, however, been shown to counteract thrombosis and to inhibit thrombin-induced production of plasminogen activator inhibitor-1, and endothelial adhesion molecule expression [1]. "
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