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Am J Physiol Cell Physiol 283: C1469-C1479, 2002. First published June 20, 2002; doi:10.1152/ajpcell.00114.2002
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Vol. 283, Issue 5, C1469-C1479, November 2002

Lyn- and ERK-mediated vs. Ca2+-mediated neutrophil O<UP><SUB>2</SUB><SUP>−</SUP></UP> responses with thermal injury

Nadeem Fazal, Walid M. Al-Ghoul, Megan J. Schmidt, Mashkoor A. Choudhry, and Mohammed M. Sayeed

Burn & Shock Trauma Institute, Department of Surgery, Stritch School of Medicine, Loyola University Chicago, Maywood, Illinois 60153

We evaluated the dependency of neutrophil O<UP><SUB>2</SUB><SUP>−</SUP></UP> production on PTK-Lyn and MAPK-ERK1/2 in rats after thermal injury. Activation of PTK-Lyn was assessed by immunoprecipitation. Phosphorylation of ERK1/2 was assessed by Western blot analysis. O<UP><SUB>2</SUB><SUP>−</SUP></UP> production was measured by isoluminol-enhanced luminometry. Imaging technique was employed to measure neutrophil [Ca2+]i in individual cells. Thermal injury caused marked upregulation of Lyn and ERK1/2 accompanying enhanced neutrophil O<UP><SUB>2</SUB><SUP>−</SUP></UP> production. Treatment of rats with PTK blocker (AG556) or MAPK blocker (AG1478) before burn injury caused complete inhibition of the respective kinase activation. Both AG556 and AG1478 produced an ~66% inhibition in O<UP><SUB>2</SUB><SUP>−</SUP></UP> production. Treatment with diltiazem (DZ) produced an ~37% inhibition of O<UP><SUB>2</SUB><SUP>−</SUP></UP> production without affecting Lyn or ERK1/2 activation with burn injury. Ca2+ mobilization was upregulated with burn injury but not affected by treatment of burn rats with AG556. Unlike the partial inhibition of burn-induced O<UP><SUB>2</SUB><SUP>−</SUP></UP> production by AG556, AG1478, or DZ, platelet-activating factor antagonist (PAFa) treatment of burn rats produced near complete inhibition of O<UP><SUB>2</SUB><SUP>−</SUP></UP> production. PAFa treatment also blocked activation of Lyn. The findings suggest that the near complete inhibition of O<UP><SUB>2</SUB><SUP>−</SUP></UP> production by PAFa was a result of blockade of PTK as well as Ca2+ signaling. Overall, our studies show that enhanced neutrophil O<UP><SUB>2</SUB><SUP>−</SUP></UP> production after thermal injury is a result of potentiation of Ca2+-linked and -independent signaling triggered by inflammatory agents such as PAF.

burn; rat; polymorphonuclear neutrophil; protein kinase C signaling; platelet-activating factor blockade; Lyn blockade; extracellular signal-regulated kinase 1/2 blockade


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