Glutathione regulates integrin α_IIbβ₃-mediated cell adhesion under flow conditions

Summary

The platelet integrin α_IIbβ₃ mediates the final step of platelet aggregation that requires pre-activation through an inside-out signal initiated by agonists. Experiments conducted under static conditions using platelet-rich plasma show that platelet activation and adhesion activity of α_IIbβ₃ are regulated by glutathione (GSH-GSSG) redox potential.However,it remains unclear as to whether GSH-GSSG exerts its regulatory role in platelets by direct targeting of α_IIbβ₃ or intracellular signals that activate the integrin. A role of fluid shear stress is also not known. We examined the effects of GSH-GSSG on the adhesion of CHO cells expressing two HPA variants of human α_IIbβ₃ to the immobilized fibrinogen and von Willebrand factor (VWF) under flow conditions. GSH-GSSG dose-dependently reduced the number of adherent cells to fibrinogen and VWF under 2.5 dyn/cm² of shear stress, a physical force calculated to be 110 dyne on platelets. GSH treatment also abolished the hyperadhesion activity of cells expressing the Pro33 variant of α_IIbβ₃.The inhibition was also observed with washed platelets. The data differ from the early observation that GSH enhanced platelet aggregation induced by sub-threshold concentrations of platelet agonists. The results suggest that GSH may have distinct effects on agonist-induced α_IIbβ₃ activation and on the α_IIbβ₃-fibrinogen or α_IIbβ₃-VWF bonds when exposed to fluid shear stress. They further suggest that the HPA phenotype may be redox-regulated.

Keywords

Adhesion receptors - platelet glycoproteins - rheology / mechanical stimuli - fibrinogen / fibrin - von Willebrand factor

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