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 language | English |
|---|---|
| Pages (from-to) | 695-701 |
| Number of pages | 7 |
| Journal | British Journal of Pharmacology |
| Volume | 120 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 1997 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - NO/PGI2-independent vasorelaxation and the cytochrome P450 pathway in rabbit carotid artery
AU - Dong, Hui
AU - Waldron, Gareth J.
AU - Galipeau, Denise
AU - Cole, William C.
AU - Triggle, Christopher
PY - 1997
Y1 - 1997
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.
KW - Acetylcholine
KW - Ca-activated K channels
KW - Carotid artery
KW - Cytochrome P450
KW - Endothelium-derived hyperpolarizing factor (EDHF)
KW - NO
KW - Vasorelaxation
UR - http://www.scopus.com/inward/record.url?scp=0030614947&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030614947&partnerID=8YFLogxK
U2 - 10.1038/sj.bjp.0700945
DO - 10.1038/sj.bjp.0700945
M3 - Article
C2 - 9051310
AN - SCOPUS:0030614947
VL - 120
SP - 695
EP - 701
JO - British Journal of Pharmacology
JF - British Journal of Pharmacology
SN - 0007-1188
IS - 4
ER -