Role of CYP epoxygenases in A2AAR-mediated relaxation using A2AAR-null and wild-type mice

Mohammed A. Nayeem, Samuel M. Poloyac, John R. Falck, Darryl C. Zeldin, Catherine Ledent, Dovenia S. Ponnoth, Habib R. Ansari, S. Jamal Mustafa

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

We hypothesized that A2A adenosine receptor (A2AAR) activation causes vasorelaxation through cytochrome P-450 (CYP) epoxygenases and endothelium-derived hyperpolarizing factors, whereas lack of A2AAR activation promotes vasoconstriction through Cyp4a in the mouse aorta. Adenosine 5′-N-ethylcarboxamide (NECA; 10-6 M), an adenosine analog, caused relaxation in wild-type A2AAR (A2AAR+/+; +33.99 ± 4.70%, P < 0.05) versus contraction in A2AAR knockout (A2AAR-/-; -27.52 ± 4.11%) mouse aortae. An A2AAR-specific antagonist (SCH-58261; 1μ-M) changed the NECA (10-6 M) relaxation response to contraction (-35.82 ± 4.69%, P < 0.05) in A2AAR+/+ aortae, whereas no effect was noted in A2AAR-/- aortae. Significant contraction was seen in the absence of the endothelium in A2AAR+/+ (-2.58 ± 2.25%) aortae compared with endothelium-intact aortae. An endothelial nitric oxide synthase inhibitor (N-nitro-L-arginine methyl ester; 100 μ-M) and a cyclooxygenase inhibitor (indomethacin; 10 μ-M) failed to block NECA-induced relaxation in A2AAR+/+ aortae. A selective inhibitor of CYP epoxygenases (methylsulfonyl-propargyloxyphenylhexanamide; 10 -μM) changed NECA-mediated relaxation (-22.74 ± 5.11% at 10 -6 M) and CGS-21680-mediated relaxation (-18.54 ± 6.06% at 10-6 M) to contraction in A2AAR+/+ aortae, whereas no response was noted in A2AAR-/- aortae. Furthermore, an epoxyeicosatrienoic acid (EET) antagonist [14,15-epoxyeicosa- 5(Z)-enoic acid; 10 μ-M] was able to block NECA-induced relaxation in A 2AAR+/+ aortae, whereas ω-hydroxylase inhibitors (10 μ-M dibromo-dodecenyl-methylsulfimide and 10 -μM HET-0016) changed contraction into relaxation in A2AAR-/- aorta. Cyp2c29 protein was upregulated in A2AAR+/+ aortae, whereas Cyp4a was upregulated in A2AAR-/- aortae. Higher levels of dihydroxyeicosatrienoic acids (DHETs; 14,15-DHET, 11,12-DHET, and 8,9-DHET, P < 0.05) were found in A2AAR+/+ versus A 2AAR-/- aortae. EET levels were not significantly different between A2AAR+/+ and A2AAR -/- aortae. It is concluded that CYP epoxygenases play an important role in A2AAR-mediated relaxation, and the deletion of the A 2AAR leads to contraction through Cyp4a.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume295
Issue number5
DOIs
StatePublished - Nov 2008

Fingerprint

Adenosine A2A Receptors
Cytochrome P-450 Enzyme System
Aorta
Adenosine-5'-(N-ethylcarboxamide)
Endothelium
Acids
Adenosine A2 Receptor Antagonists

Keywords

  • Adenosine
  • Cytochrome P-450s
  • Dihydroxyeicosatrienoic acids
  • Epoxyeicosatrienoic acids
  • Vasoconstriction
  • Vasodilation

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)
  • Cardiology and Cardiovascular Medicine

Cite this

Role of CYP epoxygenases in A2AAR-mediated relaxation using A2AAR-null and wild-type mice. / Nayeem, Mohammed A.; Poloyac, Samuel M.; Falck, John R.; Zeldin, Darryl C.; Ledent, Catherine; Ponnoth, Dovenia S.; Ansari, Habib R.; Mustafa, S. Jamal.

In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 295, No. 5, 11.2008.

Research output: Contribution to journalArticle

Nayeem, Mohammed A. ; Poloyac, Samuel M. ; Falck, John R. ; Zeldin, Darryl C. ; Ledent, Catherine ; Ponnoth, Dovenia S. ; Ansari, Habib R. ; Mustafa, S. Jamal. / Role of CYP epoxygenases in A2AAR-mediated relaxation using A2AAR-null and wild-type mice. In: American Journal of Physiology - Heart and Circulatory Physiology. 2008 ; Vol. 295, No. 5.
@article{8c5bdc26899f4d7c84c2b38f33740bca,
title = "Role of CYP epoxygenases in A2AAR-mediated relaxation using A2AAR-null and wild-type mice",
abstract = "We hypothesized that A2A adenosine receptor (A2AAR) activation causes vasorelaxation through cytochrome P-450 (CYP) epoxygenases and endothelium-derived hyperpolarizing factors, whereas lack of A2AAR activation promotes vasoconstriction through Cyp4a in the mouse aorta. Adenosine 5′-N-ethylcarboxamide (NECA; 10-6 M), an adenosine analog, caused relaxation in wild-type A2AAR (A2AAR+/+; +33.99 ± 4.70{\%}, P < 0.05) versus contraction in A2AAR knockout (A2AAR-/-; -27.52 ± 4.11{\%}) mouse aortae. An A2AAR-specific antagonist (SCH-58261; 1μ-M) changed the NECA (10-6 M) relaxation response to contraction (-35.82 ± 4.69{\%}, P < 0.05) in A2AAR+/+ aortae, whereas no effect was noted in A2AAR-/- aortae. Significant contraction was seen in the absence of the endothelium in A2AAR+/+ (-2.58 ± 2.25{\%}) aortae compared with endothelium-intact aortae. An endothelial nitric oxide synthase inhibitor (N-nitro-L-arginine methyl ester; 100 μ-M) and a cyclooxygenase inhibitor (indomethacin; 10 μ-M) failed to block NECA-induced relaxation in A2AAR+/+ aortae. A selective inhibitor of CYP epoxygenases (methylsulfonyl-propargyloxyphenylhexanamide; 10 -μM) changed NECA-mediated relaxation (-22.74 ± 5.11{\%} at 10 -6 M) and CGS-21680-mediated relaxation (-18.54 ± 6.06{\%} at 10-6 M) to contraction in A2AAR+/+ aortae, whereas no response was noted in A2AAR-/- aortae. Furthermore, an epoxyeicosatrienoic acid (EET) antagonist [14,15-epoxyeicosa- 5(Z)-enoic acid; 10 μ-M] was able to block NECA-induced relaxation in A 2AAR+/+ aortae, whereas ω-hydroxylase inhibitors (10 μ-M dibromo-dodecenyl-methylsulfimide and 10 -μM HET-0016) changed contraction into relaxation in A2AAR-/- aorta. Cyp2c29 protein was upregulated in A2AAR+/+ aortae, whereas Cyp4a was upregulated in A2AAR-/- aortae. Higher levels of dihydroxyeicosatrienoic acids (DHETs; 14,15-DHET, 11,12-DHET, and 8,9-DHET, P < 0.05) were found in A2AAR+/+ versus A 2AAR-/- aortae. EET levels were not significantly different between A2AAR+/+ and A2AAR -/- aortae. It is concluded that CYP epoxygenases play an important role in A2AAR-mediated relaxation, and the deletion of the A 2AAR leads to contraction through Cyp4a.",
keywords = "Adenosine, Cytochrome P-450s, Dihydroxyeicosatrienoic acids, Epoxyeicosatrienoic acids, Vasoconstriction, Vasodilation",
author = "Nayeem, {Mohammed A.} and Poloyac, {Samuel M.} and Falck, {John R.} and Zeldin, {Darryl C.} and Catherine Ledent and Ponnoth, {Dovenia S.} and Ansari, {Habib R.} and Mustafa, {S. Jamal}",
year = "2008",
month = "11",
doi = "10.1152/ajpheart.01333.2007",
language = "English (US)",
volume = "295",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "5",

}

TY - JOUR

T1 - Role of CYP epoxygenases in A2AAR-mediated relaxation using A2AAR-null and wild-type mice

AU - Nayeem, Mohammed A.

AU - Poloyac, Samuel M.

AU - Falck, John R.

AU - Zeldin, Darryl C.

AU - Ledent, Catherine

AU - Ponnoth, Dovenia S.

AU - Ansari, Habib R.

AU - Mustafa, S. Jamal

PY - 2008/11

Y1 - 2008/11

N2 - We hypothesized that A2A adenosine receptor (A2AAR) activation causes vasorelaxation through cytochrome P-450 (CYP) epoxygenases and endothelium-derived hyperpolarizing factors, whereas lack of A2AAR activation promotes vasoconstriction through Cyp4a in the mouse aorta. Adenosine 5′-N-ethylcarboxamide (NECA; 10-6 M), an adenosine analog, caused relaxation in wild-type A2AAR (A2AAR+/+; +33.99 ± 4.70%, P < 0.05) versus contraction in A2AAR knockout (A2AAR-/-; -27.52 ± 4.11%) mouse aortae. An A2AAR-specific antagonist (SCH-58261; 1μ-M) changed the NECA (10-6 M) relaxation response to contraction (-35.82 ± 4.69%, P < 0.05) in A2AAR+/+ aortae, whereas no effect was noted in A2AAR-/- aortae. Significant contraction was seen in the absence of the endothelium in A2AAR+/+ (-2.58 ± 2.25%) aortae compared with endothelium-intact aortae. An endothelial nitric oxide synthase inhibitor (N-nitro-L-arginine methyl ester; 100 μ-M) and a cyclooxygenase inhibitor (indomethacin; 10 μ-M) failed to block NECA-induced relaxation in A2AAR+/+ aortae. A selective inhibitor of CYP epoxygenases (methylsulfonyl-propargyloxyphenylhexanamide; 10 -μM) changed NECA-mediated relaxation (-22.74 ± 5.11% at 10 -6 M) and CGS-21680-mediated relaxation (-18.54 ± 6.06% at 10-6 M) to contraction in A2AAR+/+ aortae, whereas no response was noted in A2AAR-/- aortae. Furthermore, an epoxyeicosatrienoic acid (EET) antagonist [14,15-epoxyeicosa- 5(Z)-enoic acid; 10 μ-M] was able to block NECA-induced relaxation in A 2AAR+/+ aortae, whereas ω-hydroxylase inhibitors (10 μ-M dibromo-dodecenyl-methylsulfimide and 10 -μM HET-0016) changed contraction into relaxation in A2AAR-/- aorta. Cyp2c29 protein was upregulated in A2AAR+/+ aortae, whereas Cyp4a was upregulated in A2AAR-/- aortae. Higher levels of dihydroxyeicosatrienoic acids (DHETs; 14,15-DHET, 11,12-DHET, and 8,9-DHET, P < 0.05) were found in A2AAR+/+ versus A 2AAR-/- aortae. EET levels were not significantly different between A2AAR+/+ and A2AAR -/- aortae. It is concluded that CYP epoxygenases play an important role in A2AAR-mediated relaxation, and the deletion of the A 2AAR leads to contraction through Cyp4a.

AB - We hypothesized that A2A adenosine receptor (A2AAR) activation causes vasorelaxation through cytochrome P-450 (CYP) epoxygenases and endothelium-derived hyperpolarizing factors, whereas lack of A2AAR activation promotes vasoconstriction through Cyp4a in the mouse aorta. Adenosine 5′-N-ethylcarboxamide (NECA; 10-6 M), an adenosine analog, caused relaxation in wild-type A2AAR (A2AAR+/+; +33.99 ± 4.70%, P < 0.05) versus contraction in A2AAR knockout (A2AAR-/-; -27.52 ± 4.11%) mouse aortae. An A2AAR-specific antagonist (SCH-58261; 1μ-M) changed the NECA (10-6 M) relaxation response to contraction (-35.82 ± 4.69%, P < 0.05) in A2AAR+/+ aortae, whereas no effect was noted in A2AAR-/- aortae. Significant contraction was seen in the absence of the endothelium in A2AAR+/+ (-2.58 ± 2.25%) aortae compared with endothelium-intact aortae. An endothelial nitric oxide synthase inhibitor (N-nitro-L-arginine methyl ester; 100 μ-M) and a cyclooxygenase inhibitor (indomethacin; 10 μ-M) failed to block NECA-induced relaxation in A2AAR+/+ aortae. A selective inhibitor of CYP epoxygenases (methylsulfonyl-propargyloxyphenylhexanamide; 10 -μM) changed NECA-mediated relaxation (-22.74 ± 5.11% at 10 -6 M) and CGS-21680-mediated relaxation (-18.54 ± 6.06% at 10-6 M) to contraction in A2AAR+/+ aortae, whereas no response was noted in A2AAR-/- aortae. Furthermore, an epoxyeicosatrienoic acid (EET) antagonist [14,15-epoxyeicosa- 5(Z)-enoic acid; 10 μ-M] was able to block NECA-induced relaxation in A 2AAR+/+ aortae, whereas ω-hydroxylase inhibitors (10 μ-M dibromo-dodecenyl-methylsulfimide and 10 -μM HET-0016) changed contraction into relaxation in A2AAR-/- aorta. Cyp2c29 protein was upregulated in A2AAR+/+ aortae, whereas Cyp4a was upregulated in A2AAR-/- aortae. Higher levels of dihydroxyeicosatrienoic acids (DHETs; 14,15-DHET, 11,12-DHET, and 8,9-DHET, P < 0.05) were found in A2AAR+/+ versus A 2AAR-/- aortae. EET levels were not significantly different between A2AAR+/+ and A2AAR -/- aortae. It is concluded that CYP epoxygenases play an important role in A2AAR-mediated relaxation, and the deletion of the A 2AAR leads to contraction through Cyp4a.

KW - Adenosine

KW - Cytochrome P-450s

KW - Dihydroxyeicosatrienoic acids

KW - Epoxyeicosatrienoic acids

KW - Vasoconstriction

KW - Vasodilation

UR - http://www.scopus.com/inward/record.url?scp=57049160502&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=57049160502&partnerID=8YFLogxK

U2 - 10.1152/ajpheart.01333.2007

DO - 10.1152/ajpheart.01333.2007

M3 - Article

VL - 295

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 5

ER -