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Microparticles (MPs), small membrane-bound particles originating from different cell types during activation or apoptosis, mediate intercellular communication, exert pro-coagulant activity and affect inflammation and other pathophysiological conditions. Monocyte-derived MPs have undergone little investigation and, to our knowledge, have never been evaluated for their possible autocrine effects. Therefore, we assessed the ability of monocyte-derived MPs to stimulate human monocytes and monocyte-derived macrophages (MDM).MPs were generated from supernatants of human monocytes stimulated by the calcium ionophore A23187 (12 µM), and then characterized. Human monocytes and MDM of healthy donors were isolated by standard procedures. Cells were challenged by MPs or phorbol 12-myristate 13-acetate (PMA, used as standard stimulus), in the absence or presence of PPARγ agonists and antagonists. Superoxide anion production (measured spectrophotometrically), cytokine release (elisa), PPARγ protein expression (immunoblotting) and NF-κB activation (EMSA assay) were evaluated.Monocyte-derived MPs induced, in a concentration-dependent manner, oxygen radical production, cytokine release and NF-κB activation in human monocytes and macrophages, with lower effects than PMA. In both cell types, the PPARγ agonists rosiglitazone and 15-deoxy-Δ(12,14) -prostaglandin J(2) (15d-PGJ(2) ) inhibited MPs-induced stimulation and this inhibition was reversed by a PPARγ antagonist. In human monocyte/macrophages, MPs as well as rosiglitazone and 15d-PGJ(2) induced PPARγ protein expression.In human monocyte/macrophages, monocyte-derived MPs exert an autocrine activation that was modulated by PPARγ ligands, inducing both pro-inflammatory (superoxide anion production, cytokine release and NF-κB activation) and anti-inflammatory (PPARγ expression) effects.

Our previous study on human skeletal muscle undergoing ischemia and reperfusion has revealed that granulocytes, which infiltrate the muscle tissue in large numbers, play an important role in mediating fibre injuries by producing superoxide anion (O?) which is responsible for membrane lipid peroxidation. In the current study, five patients undergoing aortic reconstructive surgery were given acetyl-carnitine (2 mglkg i.v. plus 1 mglkglmin for 30 min) prior to the induction of ischemia. Muscle biopsies and blood samples were examined: a) after anaesthesia; b) at the end of ischemia; and c) 30 min after reperfusion, with the aim of elucidating whether acetylcarnitine could prevent the infiltration andlor the activation of granulocytes and eventually skeletal muscle injuries. During ischemia and reperfusion complement activation recmited numerous granulocytes into the muscle tissue, but, contrary to the untreated samples, the ability for O2 -generation of these cells remained at low levels and was comparable to that of ischemia even when molecular 02 was reintroduced to the tissue. Accordingly, the morphological changes of the postischemic muscle fibers were substantially reduced when compared to the untreated samples; in fact, the mitochondrial swelling was only moderate and the intramitochondrial dense bodies were small and scarce. The current findings support a positive role of acetyl-carnitine in ameliorating the ischernia-reperfusion (1-R)-induced damage of human skeletal muscle.

A fixed concentration of paf-acether (platelet-activating factor; 4 microM) relaxed isolated guinea pig tracheal preparations which had been contracted with histamine (50 microM), serotonin (1 microM) or leukotriene D4 (0.1 microM). The relaxations were approximately 43, 100 and 57%, respectively. We did not observe any relaxant effect of paf-acether (4 microM) in tissues contracted with acetylcholine (50 microM). Both lyso paf-acether (10 microM) and bovine serum albumin (25 micrograms/ml) were without effect on histamine-contracted preparations. In the presence of indomethacin (1.7 microM; 30 min) or aspirin (0.1 mM; 30 min) the relaxant effect of paf-acether (4 microM) in tissues contracted with histamine was significantly reduced to approximately 10 and 12%, respectively. When paf-acether (4 microM) was added to histamine-contracted tracheal preparations in the presence of noradrenaline (0.1 microM) or prostaglandin E2 (PGE2, 10 nM) the relaxations were 62 and 82%, respectively. Noradrenaline and PGE2 alone had only a slight relaxant effect in these tissues (7 and 14%, respectively). In the presence of indomethacin (1.7 microM) the synergistic effect of paf-acether and PGE2 was still observed. The basal production of PGE2 in isolated guinea pig tracheal preparations was 4.6 +/- 1.4 pg/mg of tissue. In the presence of paf-acether (4 microM) increased levels of this prostanoid were detected (11.2 +/- 2.4 pg/mg of tissue). Isolated guinea pig tracheal preparations when contracted with histamine released PGE2 (17.6 +/- 4.1 pg/mg of tissue). In the presence of histamine and paf-acether there was a significant increase in detectable levels of PGE2 (48.6 +/- 13.2 pg/mg of tissue).(ABSTRACT TRUNCATED AT 250 WORDS)