The biological actions of 11,12-epoxyeicosatrienoic acid in endothelial cells are specific to the R/S-enantiomer and require the Gs proteins

Yindi Ding, Timo Frömel, Rüdiger Popp, J R Falck, Wolf Hagen Schunck, Ingrid Fleming

Research output: Contribution to journalArticle

Abstract

Cytochrome P450-derived epoxides of arachidonic acid [i.e., the epoxyeicosatrienoic acids (EETs)] are important lipid signaling molecules involved in the regulation of vascular tone and angiogenesis. Because many actions of 11,12-cis-epoxyeicosatrienoic acid (EET) are dependent on the activation of protein kinase A (PKA), the existence of a cell-surface G s-coupled receptor has been postulated. To assess whether the responses of endothelial cells to 11,12-EET are enantiomer specific and linked to a potential G protein-coupled receptor, we assessed 11,12-EETinduced, PKA-dependent translocation of transient receptor potential (TRP) C6 channels, as well as angiogenesis. In primary cultures of human endothelial cells, (±)-11,12-EET led to the rapid (30 seconds) translocation a TRPC6-V5 fusion protein, an effect reproduced by 11(R),12(S)-EET, but not by 11(S),12(R)-EET or (±)-14,15-EET. Similarly, endothelial cell migration and tube formation were stimulated by (±)-11,12-EET and 11(R),12(S)- EET, whereas 11(S),12(R)-EET and 11,12-dihydroxyeicosatrienoic acid were without effect. The effects of (±)-11,12-EET on TRP channel translocation and angiogenesis were sensitive to EET antagonists, and TRP channel trafficking was also prevented by a PKA inhibitor. The small interfering RNA-mediated downregulation of Gs in endothelial cells had no significant effect on responses stimulated by vascular endothelial growth or a PKA activator but abolished responses to (±)-11,12-EET. The downregulation of G q/11 failed to prevent 11,12-EET-induced TRPC6 channel translocation or the formation of capillary-like structures. Taken together, our results suggest that a Gs-coupled receptor in the endothelial cell membrane responds to 11(R),12(S)-EET and mediates the PKA-dependent translocation and activation of TRPC6 channels, as well as angiogenesis.

Original languageEnglish (US)
Pages (from-to)14-21
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume350
Issue number1
DOIs
StatePublished - Jan 1 2014

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Endothelial Cells
Cyclic AMP-Dependent Protein Kinases
Proteins
Transient Receptor Potential Channels
NSC 153174
Blood Vessels
11,12-epoxy-5,8,14-eicosatrienoic acid
Down-Regulation
Gastrin-Secreting Cells
Acids
Epoxy Compounds
Protein Kinase Inhibitors
G-Protein-Coupled Receptors
Arachidonic Acid
Cytochrome P-450 Enzyme System
Small Interfering RNA
Cell Movement
Cell Membrane
Lipids
Growth

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

Cite this

The biological actions of 11,12-epoxyeicosatrienoic acid in endothelial cells are specific to the R/S-enantiomer and require the Gs proteins. / Ding, Yindi; Frömel, Timo; Popp, Rüdiger; Falck, J R; Schunck, Wolf Hagen; Fleming, Ingrid.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 350, No. 1, 01.01.2014, p. 14-21.

Research output: Contribution to journalArticle

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AU - Frömel, Timo

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AU - Falck, J R

AU - Schunck, Wolf Hagen

AU - Fleming, Ingrid

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