Evaluation of cytochrome P450 4 family as mediator of phospholipase D activation in aortic vascular smooth muscle cells

Jean Hugues Parmentier, Eduard N. Lavrentyev, John R. Falck, Jorge H. Capdevila, Kafait Malik

Research output: Contribution to journalArticle

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Abstract

Norepinephrine (NE) stimulates phospholipase D (PLD) activity via phospholipase A2-dependent arachidonic acid release in rabbit aortic vascular smooth muscle cells (VSMC). We have previously shown that exogenous 20-hydroxyeicosatetraenoic acid (20-HETE), an eicosanoid generated through the cytochrome P450 (CYP) 4A pathway in vivo, stimulates PLD activity. Whether endogenous CYP4-derived arachidonic acid metabolites act as intracellular mediators of NE-induced PLD activation in VSMC is not known. In rabbit aortic VSMC, prototypical hepatic/renal CYP4A inducers such as fenofibrate and Wy 14643 inhibited both basal and NE-induced PLD activity after 48 h of exposure. The level of CYP4F, and to a lesser extent CYP4A, was also decreased by these agents. The expression levels of rabbit aortic VSMC CYP4A and CYP4F isoforms were reduced by antisense oligonucleotides treatment for 48 hours as measured by RTQ-PCR or Western blotting. This reduction in CYP4A or CYP4F levels did not change NE-induced PLD activation. The corresponding CYP4A scrambled and CYP4F sense oligonucleotides did not alter CYP levels. PLD activity was increased by ∼70% after 15 min of stimulation with NE, whereas lauric acid ω-hydroxylase activity, a measure of fatty acid ω-hydroxylation, was unchanged. Inhibition of ω-hydroxylation with DDMS and HET0016, selective ω-hydroxylase inhibitors, and 20-HEDE, an antagonist of 20-HETE, increased PLD activity in a concentration-dependent manner and did not alter NE-induced PLD activation. These data suggest that PLD activation by NE is independent of the CYP4A/4F enzymes in rabbit aortic VSMC.

Original languageEnglish (US)
Pages (from-to)1015-1029
Number of pages15
JournalLife Sciences
Volume77
Issue number9
DOIs
StatePublished - Jul 15 2005

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Phospholipase D
Cytochrome P-450 CYP4A
Vascular Smooth Muscle
Cytochrome P-450 Enzyme System
Smooth Muscle Myocytes
Muscle
Chemical activation
Cells
Norepinephrine
Rabbits
Hydroxylation
Arachidonic Acid
Fenofibrate
Cytochrome P450 Family 4
Eicosanoids
Antisense Oligonucleotides
Phospholipases A2
Metabolites
Mixed Function Oxygenases
Oligonucleotides

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Evaluation of cytochrome P450 4 family as mediator of phospholipase D activation in aortic vascular smooth muscle cells. / Parmentier, Jean Hugues; Lavrentyev, Eduard N.; Falck, John R.; Capdevila, Jorge H.; Malik, Kafait.

In: Life Sciences, Vol. 77, No. 9, 15.07.2005, p. 1015-1029.

Research output: Contribution to journalArticle

Parmentier, Jean Hugues ; Lavrentyev, Eduard N. ; Falck, John R. ; Capdevila, Jorge H. ; Malik, Kafait. / Evaluation of cytochrome P450 4 family as mediator of phospholipase D activation in aortic vascular smooth muscle cells. In: Life Sciences. 2005 ; Vol. 77, No. 9. pp. 1015-1029.
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