Changes of soluble CD40 ligand in the progression of acute myocardial infarction associate to endothelial nitric oxide synthase polymorphisms and vascular endothelial growth factor but not to platelet CD62P expression

Abstract

Abbreviations: AMI ¼ acute myocardial infarction; APC ¼ allophycocyanin; Asp ¼ aspartate; cTNT ¼ cardiac troponin; CAD ¼ coronary artery disease; CRP ¼ C-reactive protein; CK ¼ creatine kinase; eNOS ¼ endothelial nitric oxide synthase; ELISA ¼ enzyme-linked immunosorbent assay; FITC ¼ fluorescein isothiocyanate; FAU ¼ fluorescence arbitrary units; Glu ¼ glutamic acid; LME ¼ linear mixed effects model; MPs ¼ microparticles; NO ¼ nitric oxide; NT-proBNP ¼ N-terminal pro-brain natriuretic peptide; PCI ¼ percutaneous coronary intervention; PBS ¼ phosphate-buffered saline; PE ¼ Phycoerythrin; CD62P ¼ P-selectin; sCD40L ¼ soluble CD40 ligand; SA ¼ stable angina pectoris; VEGF ¼ vascular endothelial growth factor INTRODUCTION CD40L is a signaling molecule, 1-3 implicated in thrombosis and inflammatory response to vascular injury. [4][5][6] The relationship of CD40L with coronary artery disease (CAD) has been established, 2,7-9 as also its implication in endothelial dysfunction. [10][11][12][13][14] However, whether the soluble CD40 ligand (sCD40L) could also influence endothelial dysfunction after acute myocardial infarction (AMI) injury remains unclear.In vitro studies have shown that sCD40L inhibits angiogenesis and also growth factor-induced human umbilical vein endothelial cell migration, which is achieved by generation of free radicals and inhibition of nitric oxide (NO) production. 10 The authors hypothesized that the sCD40L could inhibit reendothelialization of an injured vessel, thereby affecting the restenosis. 10Research efforts have been directed toward the finding of biomarkers to assess endothelial function and its correlation with AMI. Genetic indicators, such as the polymorphisms of endothelial NO synthase (eNOS) gene, 15,16 may provide insight into endothelial cells function.Vascular endothelial growth factor (VEGF) is a wellknown promoter of angiogenesis and an endogenous regulator of endothelial integrity. [17][18][19] The prognostic information provided by VEGF independently of other markers likely points toward an important role for angiogenesis in regulating myocardial repair and reperfusion after AMI. 17,20 Current opinion suggests a differential role of CD40L (both soluble and membrane-bound forms, which includes microparticles in circulation) 21 at different stages of CAD, contrasting with the traditional view of an unvarying function of the CD40L-CD40-sCD40L system interactions in the disease. 6 In that perspective, no clear indication of the interplay of CD40L with endothelial and vascular function markers and their importance in the pathophysiology of the AMI has been obtained so far in human clinical studies. Therefore, the aim of this study was to evaluate the relationship of sCD40L with markers of platelet activation, endothelial and vascular function during an early recovery period after AMI. To achieve this goal, the time changes over 1 month of sCD40L levels were assessed in AMI patients and correlated with the CD40L expressed on platelets and microparticles, CD62P expression on platele...

Stratification of ST-elevation myocardial infarction patients based on soluble CD40L longitudinal changes

“…Previous studies using this cohort indicated that influence of medication in the values of inflammatory markers lasted for several days after PCI. 5,13 Therefore, patients' assessment 1 month after PCI represents the period of time for medication and clinical stabilization.…”

“…Having into consideration our previous exploratory analysis, 5 serum was chosen to determine sCD40L by enzyme-linked immunosorbent assay. In addition, to safeguard sCD40L stability, temperature was kept at 4 C in all steps after blood collection, that is, transport and processing.…”

“…A rigorous protocol was applied to mitigate source of error originating from pre-analytical and analytical methodologies. 5 DNA extraction and genotyping. Polymorphisms G894T and T786C of eNOS protein were analyzed.…”

“…1 Cross-sectional studies have shown circulating levels of sCD40L relate to CAD and acute manifestations of the disease, such as, acute myocardial infarction (AMI). [2][3][4][5][6] However, an unequivocal indication of its utility in chronic or acute CAD manifestations has not been obtained so far.…”

“…In a previous study conducted by us, 5 we have studied association of sCD40L with markers of endothelial and vascular function and of platelet activation, after AMI. On that cohort, evidences were founded of a stratification of patients with ST segment elevation myocardial infarction (STEMI) based on sCD40L concentrations 1 month after percutaneous coronary intervention (PCI).…”