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Kruse-Elliott Kris T.; Chaban, Kamal; Grossman, Jeffrey E.; Tomasko, Steve; Kamke, Chris; Darien, Benjamin
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Because platelet-activating factor (PAF) is an important mediator of inflammation and heparin has anti-inflammatory effects, we hypothesized that low molecular weight heparin (LMWH) would inhibit PAF-induced activation and chemotaxis in porcine neutrophils. Citrated blood was obtained from pento-barbital-anesthetized pigs, and neutrophils were isolated over a 55%/65% Percoll gradient. The effect of LMWH on basal phorbol myristate acetate (PMA)-induced superoxide (SO) release, as well as its effect on PAF priming for PMA-induced SO release, were investigated. Additionally, the effect of LMWH on PAF-induced chemotaxis of neutrophils across transwell membranes was evaluated. Baseline SO release in response to PMA was .351 ± .046 nmol/106 cells/min, and this was decreased to .289 ± .034 nmol/106 cells/min by pretreatment with 50 U/mL LMWH. PMA-induced SO production was increased by .240 ± .042 nmol/106 cells/min when cells were primed with 10 ±M PAF. This priming effect of PAF was reduced significantly by pretreatment of neutrophils with LMWH at 10 and 50 U/mL. Chemotaxis of neutrophils in response to 100 ±M PAF was significantly decreased to 70.02 ± 6.4% (n = 8) of the control response by pretreatment of cells with 50 U/mL LMWH. We conclude that LMWH has anti-inflammatory effects on porcine neutrophils, which includes attenuation of cell activation and chemotaxis in response to the lipid-derived inflammatory mediator, PAF.

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