Proteomic Screening of Cerebrovascular Diseases Markers in Activated Platelets

Abstract

Ab s t r a c tPurpose: The circulating anucleated platelets are involved in pathophysiology of several cerebrovascular diseases (CVD) such as atherosclerosis, Alzheimer and stroke. Platelets should be activated before involving in the inflammatory or hemostatic events. Oxidized low density lipoprotein (oxLDL) as oxidative stress inducer and one of the major risk factors in atherosclerosis leads to vast changes in platelets proteome. The objective here is to study proteins involved in such phenomena.Methods: In the current study, quiescent human platelets activated by oxLDL.2D SDS-PAGE (pH 3-10) were used to separate platelet proteins. Progenesis SameSpots statistical software was used to compare the gel images of resting and activated platelets with each other. Finally, comparatively expressed proteins were identified using Matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF) mass spectroscopy technique (MALDI-TOF MS). Results:This study showed altered levels of actin binding proteins such as myosin-9, Cofilin-1 and transgelin2. It has also pointed to overexpression of Thrombospondin (TSP)-1, Fibrinogen, Protooncogene tyrosine-protein kinase Src, and underexpression of Enoyl-CoA hydratase, mitochondrial, Ubiquitin-like modifier-activating enzyme 7.Conclusion: Alteration in actin associated protein hints towards the cytoskeletal changes necessary to oxidative stress effect on platelets. Additionally, altered expression levels of Thrombospondin (TSP)-1and Proto-oncogene tyrosine-protein kinase following oxidative stress signifies that cerebrovascular events can partly modulate via Src kinase pathway or/and TSP-TNFα-TLR4/NF-Kb pathway. These findings can raise our basic knowledge about platelet function and novel drug targets, leading to the discovery of the mechanism(s) of action of new antiplatelet drugs.