title: Molecular mechanism of nucleotide-induced primary granule release in human neutrophils: role for the P2Y2 receptor.
abstract: Nucleotides are released during vascular injury from activated platelets and broken cells, which could stimulate human neutrophils. In this study, we characterized the P2Y receptors and investigated the functional effects of extracellular nucleotides on human neutrophils. Pharmacological characterization using selective agonists and pertussis toxin revealed that human neutrophils express only functional P2Y2 receptors. However, P2Y2 receptor agonists ATP or uridine triphosphate (UTP) caused intracellular Ca2+ increases in isolated human neutrophils with an EC50 of 1 microM but failed to cause release of primary granules from human neutrophils. ATP and UTP were equally potent in causing elastase release from human neutrophils in the presence of exogenous soluble fibrinogen, whereas ADP and UDP were without effect. We investigated whether nucleotides depend on generated arachidonic acid metabolites to cause degranulation. However, phenidone and MK-886, inhibitors of the 5-lipoxygenase pathway, failed to block nucleotide-induced intracellular calcium mobilization and elastase release. ATP and UTP caused activation of p38 MAPK and ERK1/2 in human neutrophils. In addition, the inhibitors of the MAPK pathway, SB-203580 and U-0126, inhibited nucleotide-induced elastase release. We conclude that fibrinogen is required for nucleotide-induced primary granule release from human neutrophils through the P2Y2 receptor without a role for arachidonic acid metabolites. Both ERK1/2 and p38 MAPK play an important role in nucleotide-induced primary granule release from human neutrophils.
authors: Department of Physiology, Temple University Medical School, 3420 N. Broad St., Philadelphia, PA, 19140, USA.
authors: John
authors: J
authors: Meshki
authors: Florin
authors: F
authors: Tuluc
authors: Ovidiu
authors: O
authors: Bredetean
authors: Zhongren
authors: Z
authors: Ding
authors: Satya P
authors: SP
authors: Kunapuli
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D002070:Butadienes / Q000494:pharmacology; D015550:Cell Degranulation; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005340:Fibrinogen / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology*; D009570:Nitriles / Q000494:pharmacology; D010196:Pancreatic Elastase / Q000378:metabolism; D011720:Pyrazoles / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000502:physiology*; D058473:Receptors, Purinergic P2Y2; D014544:Uridine Triphosphate / Q000494:pharmacology*; D002722:Chlorobenzenes
chemical_list: D000255:Adenosine Triphosphate; D002070:Butadienes; D004791:Enzyme Inhibitors; D005340:Fibrinogen; D007093:Imidazoles; D007211:Indoles; D016859:Lipoxygenase Inhibitors; D020928:Mitogen-Activated Protein Kinases; D009570:Nitriles; D010196:Pancreatic Elastase; D011720:Pyrazoles; D011725:Pyridines; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; D014544:Uridine Triphosphate; C546385:P2RY2 protein, human; C113580:U 0126; C060893:MK-886; D002722:Chlorobenzenes; C015429:phenidone; C093642:SB 203580