Efecto de las plaquetas en la expresión proteica de segmentos de aorta sana y preinflamadaun abordaje proteómico

Abstract

Although much is known about the effects of the vascular wall on platelets, studies examining the effect of platelets on the expression of proteins in the vascular wall are sparse. MATERIALS AND METHODS. We used an in vitro model coincubating human platelet rich plasma (PRP) with control and TNF-#xF061;-preincubated bovine aortic segments. Two different platelet concentrations (105 and 107 platelets/well) were used. The coincubation system was prepared by placing sterile Transwell-COL(Corning incorporated) inserts, containing the platelets, into wells containing the bovine aortic segments. 2-DE, MS and western blot analysis were used to determine changes in the expression of proteins associated with the cytoskeleton and energetic metabolism in the aortic segments. The enzymatic activity was determined and the content of pyruvate was also quantified. RESULTS. In control healthy vascular wall, only the cytoskeleton-related proteins expression was modified by PRP. However, when PRP was coincubated with TNF-#xF061; pre-stimulated aortic segments lesser number of cytoskeleton-related proteins were modified. With respect to energetic metabolism, in control segments PRP failed to modify any of the analyzed energetic-related proteins. However, in TNF-#xF061;-preincubated segments the presence of PRP upexpressed glyceraldehyde-3- phosphate dehydrogenase. Moreover, by western blot experiments it was observed that in TNF-#xF061;-preincubated segments the expression of fructose 1,6-bisphosphate aldolase was downregulated by platelets. In addition, the activity of fructose 1,6-bisphosphate aldolase and piruvate content was reduced without modification on triosephosphate isomerase activity. CONCLUSIONS. In the control vascular wall platelets by themselves affected the expression of a number of proteins associated with contractile system and the energetic metabolism. However, under vascular pre-inflammatory state, platelets modified the expression of a lesser number of proteins associated with contractile system and energetic metabolism.