NO/PGI2-independent vasorelaxation and the cytochrome P450 pathway in rabbit carotid artery

Hui Dong, Gareth J. Waldron, Denise Galipeau, William C. Cole, Christopher Triggle

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

1. The nature and cellular mechanisms that are responsible for endothelium-dependent relaxations resistant to indomethacin and N(G)-nitro-L-arginine methyl ester (L-NAME) were investigated in phenylephrine (PE) precontracted isolated carotid arteries from the rabbit. 2. In the presence of the cyclo-oxygenase inhibitor, indomethacin (10 μM), acetylcholine (ACh) induced a concentration- and endothelium-dependent relaxation of PE-induced tone which was more potent than the calcium ionophore A23187 with pD2 values of 7.03 ± 0.12 (n = 8) and 6.37 ± 0.12 (n = 6), respectively. The ACh-induced response was abolished by removal of the endothelium, but was not altered when indomethacin was omitted (pD2 value 7.00 ± 0.10 and maximal relaxation 99 ± 3%, n = 6). Bradykinin and histamine (0.01-100 μM) had no effect either upon resting or PE-induced tone (n = 5). 3. In the presence of indomethacin plus the NO synthase inhibitor, L-NAME (30 μM), the response to A23187 was abolished. However, the response to ACh was not abolished, although it was significantly inhibited with the pD2 value and the maximal relaxation decreasing to 6.48 ± 0.10 and 67 ± 3%, respectively (for both P < 0.01, n = 8). The L-NAME/indomethacin insensitive vasorelaxation to ACh was completely abolished by preconstriction of the tissues with potassium chloride (40 mM, n = 8). 4. The Ca2+-activated K+ (K(Ca)) channel blockers, tetrabutylammonium (TBA, 1 mM, n = 5) and charybdotoxin (CTX, 0.1 μM, n = 5), completely inhibited the nitric oxide (NO) and prostacyclin (PGI2)-independent relaxation response to ACh. However, iberiotoxin (ITX, 0.1 μM, n = 8) or apamin (1-3 μM, n = 6) only partially inhibited the relaxation. 5. Inhibitors of the cytochrome P450 mono-oxygenase, SKF-525A (1-10 μM, n = 6), clotrimazole (1 μM, n = 5) and 17-octadecynoic acid (17-ODYA, 3 μM, n = 7) also reduced the NO/PGI2-independent relaxation response to ACh. 6. In endothelium-denuded rings of rabbit carotid arteries, the relaxation response to exogenous NO was not altered by either K(Ca) channel blockade with apamin (1 μM, n = 5) or CTX (0.1 μM, n = 5), or by the cytochrome P450 mono-oxygenase blockers SKF-525A (10 μM, n = 4) and clotrimazole (10 μM, n = 5). However, the NO-induced response was shifted to the right by LY83583 (10 μM, n = 4), a guanylyl cyclase inhibitor, with the pD2 value decreasing from 6.95 ± 0.14 to 6.04 ± 0.09 (P < 0.01). 7. ACh (0.01-100 μM) induced a concentration-dependent relaxation of PE-induced tone in endothelium-denuded arterial segments sandwiched with endothelium-intact donor segments. This relaxation to ACh was largely unaffected by indomathacin (10 μM) plus L-NAME (30 μM), but abolished by the combination of indomethacin, L-NAME and TBA (1 mM, n = 5). 8 These data suggest that in the rabbit carotid artery: (a) ACh can induce the release of both NO and EDHF, whereas A23187 only evokes the release of NO from the endothelium, (b) the diffusible EDHF released by ACh may be a cytochrome P450-derived arachidonic acid metabolite, and (c) EDHF-induced relaxation involves the opening of at least two types of K(Ca) channels, whereas NO mediates vasorelaxation via a guanosine 3':5'-cyclic monophosphate (cyclic GMP)-mediated pathway, in which a cytochrome P450 pathway and K(Ca) channels do not seem to be involved.

Original languageEnglish
Pages (from-to)695-701
Number of pages7
JournalBritish Journal of Pharmacology
Volume120
Issue number4
DOIs
Publication statusPublished - 1997
Externally publishedYes

Fingerprint

Epoprostenol
Carotid Arteries
Vasodilation
Cytochrome P-450 Enzyme System
Acetylcholine
Nitric Oxide
Rabbits
NG-Nitroarginine Methyl Ester
Endothelium
Indomethacin
Phenylephrine
Calcimycin
Proadifen
Clotrimazole
Apamin
Oxygenases
6-anilino-5,8-quinolinedione
Charybdotoxin
Cyclooxygenase Inhibitors
Potassium Chloride

Keywords

  • Acetylcholine
  • Ca-activated K channels
  • Carotid artery
  • Cytochrome P450
  • Endothelium-derived hyperpolarizing factor (EDHF)
  • NO
  • Vasorelaxation

ASJC Scopus subject areas

  • Pharmacology

Cite this

NO/PGI2-independent vasorelaxation and the cytochrome P450 pathway in rabbit carotid artery. / Dong, Hui; Waldron, Gareth J.; Galipeau, Denise; Cole, William C.; Triggle, Christopher.

In: British Journal of Pharmacology, Vol. 120, No. 4, 1997, p. 695-701.

Research output: Contribution to journalArticle

Dong, Hui ; Waldron, Gareth J. ; Galipeau, Denise ; Cole, William C. ; Triggle, Christopher. / NO/PGI2-independent vasorelaxation and the cytochrome P450 pathway in rabbit carotid artery. In: British Journal of Pharmacology. 1997 ; Vol. 120, No. 4. pp. 695-701.
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abstract = "1. The nature and cellular mechanisms that are responsible for endothelium-dependent relaxations resistant to indomethacin and N(G)-nitro-L-arginine methyl ester (L-NAME) were investigated in phenylephrine (PE) precontracted isolated carotid arteries from the rabbit. 2. In the presence of the cyclo-oxygenase inhibitor, indomethacin (10 μM), acetylcholine (ACh) induced a concentration- and endothelium-dependent relaxation of PE-induced tone which was more potent than the calcium ionophore A23187 with pD2 values of 7.03 ± 0.12 (n = 8) and 6.37 ± 0.12 (n = 6), respectively. The ACh-induced response was abolished by removal of the endothelium, but was not altered when indomethacin was omitted (pD2 value 7.00 ± 0.10 and maximal relaxation 99 ± 3{\%}, n = 6). Bradykinin and histamine (0.01-100 μM) had no effect either upon resting or PE-induced tone (n = 5). 3. In the presence of indomethacin plus the NO synthase inhibitor, L-NAME (30 μM), the response to A23187 was abolished. However, the response to ACh was not abolished, although it was significantly inhibited with the pD2 value and the maximal relaxation decreasing to 6.48 ± 0.10 and 67 ± 3{\%}, respectively (for both P < 0.01, n = 8). The L-NAME/indomethacin insensitive vasorelaxation to ACh was completely abolished by preconstriction of the tissues with potassium chloride (40 mM, n = 8). 4. The Ca2+-activated K+ (K(Ca)) channel blockers, tetrabutylammonium (TBA, 1 mM, n = 5) and charybdotoxin (CTX, 0.1 μM, n = 5), completely inhibited the nitric oxide (NO) and prostacyclin (PGI2)-independent relaxation response to ACh. However, iberiotoxin (ITX, 0.1 μM, n = 8) or apamin (1-3 μM, n = 6) only partially inhibited the relaxation. 5. Inhibitors of the cytochrome P450 mono-oxygenase, SKF-525A (1-10 μM, n = 6), clotrimazole (1 μM, n = 5) and 17-octadecynoic acid (17-ODYA, 3 μM, n = 7) also reduced the NO/PGI2-independent relaxation response to ACh. 6. In endothelium-denuded rings of rabbit carotid arteries, the relaxation response to exogenous NO was not altered by either K(Ca) channel blockade with apamin (1 μM, n = 5) or CTX (0.1 μM, n = 5), or by the cytochrome P450 mono-oxygenase blockers SKF-525A (10 μM, n = 4) and clotrimazole (10 μM, n = 5). However, the NO-induced response was shifted to the right by LY83583 (10 μM, n = 4), a guanylyl cyclase inhibitor, with the pD2 value decreasing from 6.95 ± 0.14 to 6.04 ± 0.09 (P < 0.01). 7. ACh (0.01-100 μM) induced a concentration-dependent relaxation of PE-induced tone in endothelium-denuded arterial segments sandwiched with endothelium-intact donor segments. This relaxation to ACh was largely unaffected by indomathacin (10 μM) plus L-NAME (30 μM), but abolished by the combination of indomethacin, L-NAME and TBA (1 mM, n = 5). 8 These data suggest that in the rabbit carotid artery: (a) ACh can induce the release of both NO and EDHF, whereas A23187 only evokes the release of NO from the endothelium, (b) the diffusible EDHF released by ACh may be a cytochrome P450-derived arachidonic acid metabolite, and (c) EDHF-induced relaxation involves the opening of at least two types of K(Ca) channels, whereas NO mediates vasorelaxation via a guanosine 3':5'-cyclic monophosphate (cyclic GMP)-mediated pathway, in which a cytochrome P450 pathway and K(Ca) channels do not seem to be involved.",
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AU - Dong, Hui

AU - Waldron, Gareth J.

AU - Galipeau, Denise

AU - Cole, William C.

AU - Triggle, Christopher

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N2 - 1. The nature and cellular mechanisms that are responsible for endothelium-dependent relaxations resistant to indomethacin and N(G)-nitro-L-arginine methyl ester (L-NAME) were investigated in phenylephrine (PE) precontracted isolated carotid arteries from the rabbit. 2. In the presence of the cyclo-oxygenase inhibitor, indomethacin (10 μM), acetylcholine (ACh) induced a concentration- and endothelium-dependent relaxation of PE-induced tone which was more potent than the calcium ionophore A23187 with pD2 values of 7.03 ± 0.12 (n = 8) and 6.37 ± 0.12 (n = 6), respectively. The ACh-induced response was abolished by removal of the endothelium, but was not altered when indomethacin was omitted (pD2 value 7.00 ± 0.10 and maximal relaxation 99 ± 3%, n = 6). Bradykinin and histamine (0.01-100 μM) had no effect either upon resting or PE-induced tone (n = 5). 3. In the presence of indomethacin plus the NO synthase inhibitor, L-NAME (30 μM), the response to A23187 was abolished. However, the response to ACh was not abolished, although it was significantly inhibited with the pD2 value and the maximal relaxation decreasing to 6.48 ± 0.10 and 67 ± 3%, respectively (for both P < 0.01, n = 8). The L-NAME/indomethacin insensitive vasorelaxation to ACh was completely abolished by preconstriction of the tissues with potassium chloride (40 mM, n = 8). 4. The Ca2+-activated K+ (K(Ca)) channel blockers, tetrabutylammonium (TBA, 1 mM, n = 5) and charybdotoxin (CTX, 0.1 μM, n = 5), completely inhibited the nitric oxide (NO) and prostacyclin (PGI2)-independent relaxation response to ACh. However, iberiotoxin (ITX, 0.1 μM, n = 8) or apamin (1-3 μM, n = 6) only partially inhibited the relaxation. 5. Inhibitors of the cytochrome P450 mono-oxygenase, SKF-525A (1-10 μM, n = 6), clotrimazole (1 μM, n = 5) and 17-octadecynoic acid (17-ODYA, 3 μM, n = 7) also reduced the NO/PGI2-independent relaxation response to ACh. 6. In endothelium-denuded rings of rabbit carotid arteries, the relaxation response to exogenous NO was not altered by either K(Ca) channel blockade with apamin (1 μM, n = 5) or CTX (0.1 μM, n = 5), or by the cytochrome P450 mono-oxygenase blockers SKF-525A (10 μM, n = 4) and clotrimazole (10 μM, n = 5). However, the NO-induced response was shifted to the right by LY83583 (10 μM, n = 4), a guanylyl cyclase inhibitor, with the pD2 value decreasing from 6.95 ± 0.14 to 6.04 ± 0.09 (P < 0.01). 7. ACh (0.01-100 μM) induced a concentration-dependent relaxation of PE-induced tone in endothelium-denuded arterial segments sandwiched with endothelium-intact donor segments. This relaxation to ACh was largely unaffected by indomathacin (10 μM) plus L-NAME (30 μM), but abolished by the combination of indomethacin, L-NAME and TBA (1 mM, n = 5). 8 These data suggest that in the rabbit carotid artery: (a) ACh can induce the release of both NO and EDHF, whereas A23187 only evokes the release of NO from the endothelium, (b) the diffusible EDHF released by ACh may be a cytochrome P450-derived arachidonic acid metabolite, and (c) EDHF-induced relaxation involves the opening of at least two types of K(Ca) channels, whereas NO mediates vasorelaxation via a guanosine 3':5'-cyclic monophosphate (cyclic GMP)-mediated pathway, in which a cytochrome P450 pathway and K(Ca) channels do not seem to be involved.

AB - 1. The nature and cellular mechanisms that are responsible for endothelium-dependent relaxations resistant to indomethacin and N(G)-nitro-L-arginine methyl ester (L-NAME) were investigated in phenylephrine (PE) precontracted isolated carotid arteries from the rabbit. 2. In the presence of the cyclo-oxygenase inhibitor, indomethacin (10 μM), acetylcholine (ACh) induced a concentration- and endothelium-dependent relaxation of PE-induced tone which was more potent than the calcium ionophore A23187 with pD2 values of 7.03 ± 0.12 (n = 8) and 6.37 ± 0.12 (n = 6), respectively. The ACh-induced response was abolished by removal of the endothelium, but was not altered when indomethacin was omitted (pD2 value 7.00 ± 0.10 and maximal relaxation 99 ± 3%, n = 6). Bradykinin and histamine (0.01-100 μM) had no effect either upon resting or PE-induced tone (n = 5). 3. In the presence of indomethacin plus the NO synthase inhibitor, L-NAME (30 μM), the response to A23187 was abolished. However, the response to ACh was not abolished, although it was significantly inhibited with the pD2 value and the maximal relaxation decreasing to 6.48 ± 0.10 and 67 ± 3%, respectively (for both P < 0.01, n = 8). The L-NAME/indomethacin insensitive vasorelaxation to ACh was completely abolished by preconstriction of the tissues with potassium chloride (40 mM, n = 8). 4. The Ca2+-activated K+ (K(Ca)) channel blockers, tetrabutylammonium (TBA, 1 mM, n = 5) and charybdotoxin (CTX, 0.1 μM, n = 5), completely inhibited the nitric oxide (NO) and prostacyclin (PGI2)-independent relaxation response to ACh. However, iberiotoxin (ITX, 0.1 μM, n = 8) or apamin (1-3 μM, n = 6) only partially inhibited the relaxation. 5. Inhibitors of the cytochrome P450 mono-oxygenase, SKF-525A (1-10 μM, n = 6), clotrimazole (1 μM, n = 5) and 17-octadecynoic acid (17-ODYA, 3 μM, n = 7) also reduced the NO/PGI2-independent relaxation response to ACh. 6. In endothelium-denuded rings of rabbit carotid arteries, the relaxation response to exogenous NO was not altered by either K(Ca) channel blockade with apamin (1 μM, n = 5) or CTX (0.1 μM, n = 5), or by the cytochrome P450 mono-oxygenase blockers SKF-525A (10 μM, n = 4) and clotrimazole (10 μM, n = 5). However, the NO-induced response was shifted to the right by LY83583 (10 μM, n = 4), a guanylyl cyclase inhibitor, with the pD2 value decreasing from 6.95 ± 0.14 to 6.04 ± 0.09 (P < 0.01). 7. ACh (0.01-100 μM) induced a concentration-dependent relaxation of PE-induced tone in endothelium-denuded arterial segments sandwiched with endothelium-intact donor segments. This relaxation to ACh was largely unaffected by indomathacin (10 μM) plus L-NAME (30 μM), but abolished by the combination of indomethacin, L-NAME and TBA (1 mM, n = 5). 8 These data suggest that in the rabbit carotid artery: (a) ACh can induce the release of both NO and EDHF, whereas A23187 only evokes the release of NO from the endothelium, (b) the diffusible EDHF released by ACh may be a cytochrome P450-derived arachidonic acid metabolite, and (c) EDHF-induced relaxation involves the opening of at least two types of K(Ca) channels, whereas NO mediates vasorelaxation via a guanosine 3':5'-cyclic monophosphate (cyclic GMP)-mediated pathway, in which a cytochrome P450 pathway and K(Ca) channels do not seem to be involved.

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KW - Ca-activated K channels

KW - Carotid artery

KW - Cytochrome P450

KW - Endothelium-derived hyperpolarizing factor (EDHF)

KW - NO

KW - Vasorelaxation

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