Biomedical subjects

Mario Pacileo

Publications and source records attributed to Mario Pacileo.

3 recordsLinked to original sources

PubMed2005

C-reactive protein induces tissue factor expression and promotes smooth muscle and endothelial cell proliferation.

OBJECTIVE: Inflammation plays a pivotal role in atherothrombosis. In addition to being a prognostic marker for major cardiovascular events, recent data indicate that C-reactive protein (CRP) might directly promote atherothrombosis by exerting direct effects on vascular cells. The aim of the present study was to determine whether CRP might affect the prothrombotic and proliferative characteristics of endothelial (ECs) and smooth muscle cells (SMCs). METHODS AND RESULTS: Incubation of ECs and SMCs with CRP resulted in a dose-dependent activation of cell proliferation, which was mediated by activation of the p44/42 MAP Kinase (ERK 1/2) pathway. In addition, CRP also induced tissue factor (TF) expression in both cell types in a dose-dependent fashion, exerting its effect at the transcriptional level, as demonstrated by semiquantitative and by real time PCR. Activation of the transcription factor, NF-kappaB, by CRP was demonstrated by EMSA and by suppression of TF expression by the NF-kappaB inhibitor, pyrrolidine-dithio-carbamate ammonium. CONCLUSIONS: These data indicate that CRP exerts direct effects on ECs and SMCs by promoting proliferation and TF expression and support the notion that CRP, besides representing a marker of inflammation, is an effector molecule able to induce a pro-atherothrombotic phenotype in cells of the vessel wall.

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PubMed2004

Tissue factor binding of activated factor VII triggers smooth muscle cell proliferation via extracellular signal-regulated kinase activation.

BACKGROUND: Tissue factor (TF) is the main initiator of coagulation in vivo. Recently, however, a role for TF as a cell receptor involved in signal transduction has been suggested. The aim of the present study was to assess whether activated factor VII (FVIIa) binding to TF could induce smooth muscle cell (SMC) proliferation and to clarify the possible intracellular mechanism(s) responsible for this proliferation. METHODS AND RESULTS: Cell proliferation was induced by FVIIa in a dose-dependent manner, as assessed by [3H]thymidine incorporation and direct cell counting, whereas no response was observed with active site-inhibited FVIIa (FVIIai), which is identical to FVIIa but is devoid of enzymatic activity. Similarly, no proliferation was observed when binding of FVIIa to TF was prevented by the monoclonal anti-TF antibody AP-1. Activation of the p44/42 mitogen-activated protein (MAP) kinase (extracellular signal-regulated kinases 1 and 2 [ERK 1/2]) pathway on binding of FVIIa to TF was demonstrated by transient ERK phosphorylation in Western blots and by suppression of proliferation with the specific MEK (MAP kinase/ERK kinase) inhibitor UO126. ERK phosphorylation was not observed with FVIIai or when cells were pretreated with AP-1. CONCLUSIONS: These data indicate a specific effect by which binding of FVIIa to TF on the surface of SMCs induces proliferation via a coagulation-independent mechanism and possibly indicate a new link between coagulation, inflammation, and atherosclerosis.

Animals↗

PubMed2003

Activated platelets stimulate tissue factor expression in smooth muscle cells.

UNLABELLED: Tissue factor (TF)-induced activation of the coagulation plays a key role in the pathophysiology of acute coronary syndromes. Because TF represents the initial trigger of the coagulation cascade, its expression in the arterial wall is tightly regulated. OBJECTIVE: To determine whether and which soluble mediators released during platelet activation may upregulate TF expression in smooth muscle cells (SMCs). METHODS AND RESULTS: Rabbit SMCs were challenged with collagen-activated platelets and their effects on TF mRNA transcription and protein expression were evaluated at different time points (30 min, 1, 2, 4, 8, 12 and 24 h) by RT-PCR, immunofluorescence and a two-stage colorimetric assay. A progressive increase in TF mRNA, peaking at 2 h, was evident in SMCs stimulated with activated platelets with respect to baseline. The increase in TF mRNA expression was associated with a parallel increase in TF protein, as demonstrated by immunofluorescence and by colorimetric assay. In a different set of experiments, selected platelet-derived soluble mediators were shown to induce TF mRNA expression. CONCLUSIONS: Activated platelets upregulate TF, via release of several soluble mediators, in a cell population widely expressed in the vessel wall and in atherosclerotic plaques, such as SMCs. This phenomenon might play an important role in sustaining thrombus formation in vivo.

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