Journal article
Urothelial cell platelet-activating factor production mediated by calcium-independent phospholipase A2γ
Urology (Ridgewood, N.J.), Vol.77(1), pp.248.e1-248.e7
01/2011
DOI: 10.1016/j.urology.2010.08.032
PMCID: PMC3087435
PMID: 21094990
Abstract
To determine the effect of phospholipase A(2) (PLA(2)) inhibitors on urothelial cell platelet-activating factor (PAF) production in response to tryptase stimulation.
Urothelial cells isolated from normal human ureters were immortalized with the human papillomavirus type 16E6E7 cell line (TEU-2 cells). PLA(2) activity in TEU-2 cells was measured using (16:0, [(3)H]18:1) plasmenylcholine and phosphatidylcholine substrates in the presence and absence of calcium. [(3)H]PAF production was measured in TEU-2 cells prelabeled with [(3)H] acetic acid. PAF-acetylhydrolase activity was measured by determining the amount of [(3)H] acetate hydrolyzed from [(3)H]PAF incubated with TEU-2 cellular protein. Adherence of human polymorphonuclear leukocyte (PMN) to TEU-2 cells was assessed by measuring myeloperoxidase activity in adherent PMNs after incubation with TEU-2 cells.
Most PLA(2) activity measured in TEU-2 cells was determined to be membrane-associated, calcium-independent PLA(2) and selective for plasmenylcholine substrate. Stimulation of TEU-2 cells with tryptase results in increased production of PAF and increased PMN adherence that were inhibited completely by pretreatment with the membrane-associated, calcium-independent PLA(2)γ-selective inhibitor (R)-bromoenol lactone. Pretreatment with the cytosolic PLA(2) inhibitor methyl arachidonyl fluorophosphonate resulted in potentiation of tryptase-stimulated PAF production and PMN adherence to TEU-2 cells that is a result of PAF-acetylhydrolase inhibition.
Tryptase stimulation of TEU-2 cells results in activation of membrane-associated, calcium-independent PLA(2)γ, leading to an increase in PAF production and increased PMN adherence. Inhibition of TEU-2 cell PAF-acetylhydrolase activity with methyl arachidonyl fluorophosphonate potentiated tryptase-stimulated PAF production and PMN adherence.
Details
- Title: Subtitle
- Urothelial cell platelet-activating factor production mediated by calcium-independent phospholipase A2γ
- Creators
- Prerna Rastogi - Department of Pathology, Saint Louis University School of Medicine, St. Louis, Missouri, USAAlice RickardDavid J KlumppJane McHowat
- Resource Type
- Journal article
- Publication Details
- Urology (Ridgewood, N.J.), Vol.77(1), pp.248.e1-248.e7
- Publisher
- United States
- DOI
- 10.1016/j.urology.2010.08.032
- PMID
- 21094990
- PMCID
- PMC3087435
- ISSN
- 0090-4295
- eISSN
- 1527-9995
- Grant note
- DK66119 / NIDDK NIH HHS R01 DK066119 / NIDDK NIH HHS R01 DK066119-01 / NIDDK NIH HHS
- Language
- English
- Date published
- 01/2011
- Academic Unit
- Pathology
- Record Identifier
- 9984047749302771
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