Dexamethasone enhances ATP-induced inflammatory responses in endothelial cells.
The purinergic nucleotide ATP is released from stressed cells and is implicated in vascular inflammation. Glucocorticoids are essential to stress responses and are used therapeutically, yet little information is available that describes the effects of glucocorticoids on ATP-induced inflammation. In a human microvascular endothelial cell line, extracellular ATP-induced interleukin (IL)-6 secretion in a dose- and time-dependent manner. When cells were pretreated with dexamethasone, a prototypic glucocorticoid, ATP-induced IL-6 production was enhanced in a time- and dose-dependent manner. Mifepristone, a glucocorticoid receptor antagonist, blocked these effects. ATP-induced IL-6 release was significantly inhibited by a phospholipase C inhibitor [1-[6-[((17β)-3-methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1H-pyrrole-2,5-dione (U73122)] (63.2 ± 3%, p < 0.001) and abolished by a p38 mitogen-activated protein kinase inhibitor [4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-1H-imidazole (SB 203580)] (88 ± 1%, p < 0.001). Cells treated with dexamethasone induced mRNA expression of the purinergic P2Y(2) receptor (P2Y(2)R) 1.8- ± 0.1-fold and, when stimulated with ATP, enhanced Ca(2+) release and augmented IL-6 mRNA expression. Silencing of the P2Y(2)R by its small interfering RNA decreased ATP-induced IL-6 production by 81 ± 1% (p < 0.001). Dexamethasone enhanced the transcription rate of P2Y(2)R mRNA and induced a dose-related increase in the activity of the P2Y(2)R promoter. Furthermore, dexamethasone-enhanced ATP induction of adhesion molecule transcription and augmented the release of IL-8. Dexamethasone leads to an unanticipated enhancement of endothelial inflammatory mediator production by extracellular ATP via a P2Y(2)R-dependent mechanism. These data define a novel positive feedback loop of glucocorticoids and ATP-induced endothelial inflammation.
Ding
Yi
Y
Critical Care Medicine Department, Clinical Center and Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institutes of Health, Bethesda, Maryland 20892-1662, USA.
Gao
Zhan-Guo
ZG
Jacobson
Kenneth A
KA
Suffredini
Anthony F
AF
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D020013:Calcium Signaling / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000235:genetics; D002460:Cell Line; D018932:Chemokine CCL2 / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins; D003907:Dexamethasone / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D015735:Mifepristone / Q000494:pharmacology; D009687:Nuclear Proteins / Q000235:genetics; D009711:Nucleotides / Q000494:pharmacology; D058919:Purinergic P2 Receptor Antagonists / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D011965:Receptors, Glucocorticoid / Q000037:antagonists & inhibitors / Q000378:metabolism; D058473:Receptors, Purinergic P2Y2 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013498:Suramin / Q000494:pharmacology; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism; D015854:Up-Regulation / Q000235:genetics; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
C111835:CCL2 protein, human; D015815:Cell Adhesion Molecules; D018932:Chemokine CCL2; D016207:Cytokines; D004268:DNA-Binding Proteins; C508600:IL6 protein, human; D018836:Inflammation Mediators; D015850:Interleukin-6; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; D009687:Nuclear Proteins; D009711:Nucleotides; D058919:Purinergic P2 Receptor Antagonists; D034741:RNA, Small Interfering; D011965:Receptors, Glucocorticoid; D058473:Receptors, Purinergic P2Y2; D015735:Mifepristone; D013498:Suramin; D003907:Dexamethasone; D000255:Adenosine Triphosphate; D048051:p38 Mitogen-Activated Protein Kinases; D010738:Type C Phospholipases; C076529:TNFAIP3 protein, human; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3
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