Linoleic acid enhances the secretion of plasminogen activator inhibitor type 1 by HepG2 cells.

This study was undertaken in order to assess whether triglycerides and/or their fatty acids directly influence the secretion of plasminogen activator inhibitor type 1 (PAI-1) in HepG2 cells. To this end, subconfluent HepG2 cells were incubated with triglyceride-rich particles (TGRP) isolated from Intralipid for 16 h, and PAI-1 levels were determined in conditioned medium using a specific ELISA. TGRP (1 to 6 mg triglycerides/ml) concentration-dependently increased PAI-1 secretion by cells, concomitantly with significant increases in intracellular triglyceride (TG) levels. Fatty acid analysis indicated that the incubation of cells with 3 mg of TG per ml of TGRP induced significant accumulation of 18:2 n-6 (linoleic acid, LA) and 18:3 n-3 (linolenic acid) reflecting the fatty acid composition at the added triglycerides. We then tested the comparative effects on PAI-1 secretion by HepG2 cells of LA and 18:1 n-9 (oleic acid, OA). LA, as a bovine serum albumin (BSA) complex, concentration-dependently (1 to 35 mumol/L) increased the secretion of PAI-1 by cells, whereas OA-BSA only minimally affected it at the highest concentration used (35 mumol/L). Incorporation of LA into cell pools, in the presence of increasing concentration of the FA in the medium, was studied by the use of a preparation containing [14C]LA. LA accumulated in all lipid classes including diacylglycerol, the incorporated LA being converted into arachidonic acid (AA) as assessed by HPLC radiochromatography of the fatty acid methyl esters. It is concluded that PAI-1 secretion in HepG2 cells is modulated by triacylglycerols and by linoleic acid and/or its metabolic products

Induction of plasminogen activator inhibitor I by the PPARalpha ligand, Wy-14,643, is dependent on ERK1/2 signaling pathway

Impairment of the fibrinolytic system, mostly due to elevated plasma levels of plasminogen activator inhibitor 1 (PAI-1), is often associated with metabolic disorders such as diabetes mellitus and insulin-resistance syndrome. Moreover, insulin, as we have previously shown, directly stimulates PAI-1 production with a mechanism underlying a complex signaling network which ultimately leads to ERK activation. In this study we have analyzed the effects of agonists of the peroxisome proliferator-activated receptor (PPAR) alpha and gamma on PAI-1 biosynthesis in HepG2 cells in the presence or absence of insulin. The high affinity PPARalpha agonist, Wy-14,643, increased basal and insulin-stimulated PAI-1 antigen release with a mechanism involving gene transcription. We then investigated whether the MAP kinase pathway also plays a role in the stimulatory properties of Wy-L4,643. Wy-L4,643 increases phosphorylation of ERK and p38 in a time-dependent manner without affecting that of SAPK/JNK or ERK5. Moreover, the MEK (ERK kinase) inhibitors, PD98059 and UO126, completely prevented PAI-1 induction by Wy-14,643 without inhibiting the activation of a reporter gene carrying the PPRE element. Interestingly, the addition of p38 inhibitor followed by insulin and Wy-14,643 resulted in a greater than additive stimulation of PAI-1 secretion acting through ERK1/2 phosphorylation. In contrast, the synthetic PPARgamma agonist, rosiglitazone, did not change PAI-1 level, although this compound induced transcription from the PPRE-driven luciferase reporter construct. In conclusion, Wy-14,643 induces PAI-1 gene expression, in the presence or absence of insulin, with a mechanism which is independent on PPARalpha activation and requires signaling through the ERK1/2 signaling pathway

The role of tissue factor and P-selectin in the procoagulant response that occurs in the first month after on-pump and off-pump coronary artery bypass grafting

BackgroundIt has been previously shown that a persistent (up to 1 month) prothrombotic status occurs after coronary bypass surgery performed both on pump and off pump. To assess the pathways involved in the occurrence of postoperative prothrombotic state, in this study we evaluated plasma, monocyte-bound, and platelet-bound tissue factor expression, as well as platelet and soluble P-selectin expression, up to 1 month after off-pump and on-pump coronary artery bypass grafting.MethodsThirty patient candidates for coronary surgery were randomized to undergo off-pump coronary artery bypass grafting (n = 15) or on-pump coronary artery bypass grafting (n = 15). Blood samples were collected before the intervention, after protamine administration, and 4, 8, and 30 days after surgical intervention.ResultsPlasma tissue factor levels were significantly higher than baseline both in the on-pump coronary artery bypass grafting group (from protamine administration up to 4 postoperative days) and in the off-pump coronary artery bypass grafting group (at 4 postoperative days), with no differences between groups. Basal and lipopolysaccharide-stimulated monocyte tissue factor expression, as well as basal and adenosine diphosphate–stimulated platelet tissue factor expression, did not show significant variations over time and were similar in the on-pump and off-pump coronary artery bypass grafting groups throughout the course of the study. Platelet expression of P-selectin, both basal and after adenosine diphosphate stimulation, did not significantly change over time and was not different in the on-pump and off-pump coronary artery bypass grafting groups. Soluble P-selectin levels in plasma were significantly higher in patients receiving on-pump coronary artery bypass grafting only at the time point after protamine administration, whereas this variable behaved similarly in the on-pump and off-pump coronary artery bypass grafting groups for the whole postoperative period.ConclusionsThe postoperative tissue factor and P-selectin expression did not differ between the on-pump and off-pump coronary artery bypass grafting groups. The distinct increase of plasma tissue factor occurring after both surgical procedures might represent a mechanism that might explain, in part, the early postoperative prothrombotic state occurring after on-pump and off-pump coronary artery bypass grafting