(Circulation. 1996;94:3341-3347.)
© 1996 American Heart Association, Inc.
Articles |
Nitric Oxide Attenuates the Release of Endothelium-Derived Hyperpolarizing Factor
the Zentrum der Physiologie, Klinikum der Johann Wolfgang Goethe-Universitat, Frankfurt am Main, Germany.
Correspondence to Dr Johann Bauersachs, Zentrum der Physiologie, Klinikum der Johann Wolfgang Goethe-Universitat, Theodor-Stern-Kai 7, D-60590 Frankfurt am Main, Germany.
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Abstract |
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Background The contribution of the endothelium-derived hyperpolarizing factor (EDHF), proposed to be a cytochrome P450–derived metabolite of arachidonic acid, to endothelium-dependent dilatation under physiological conditions has yet to be established, because its effect can be detected only after inhibition of NO synthase and cyclooxygenase. The possibility that NO exerts a feedback inhibition on EDHF formation was studied in isolated perfused arterial segments.
Methods and Results Under combined blockade of NO synthase and cyclooxygenase, the EDHF-mediated vasodilatation elicited by receptor-dependent agonists in rabbit carotid and porcine coronary arteries was significantly attenuated by the NO donors C87-3786 and CAS 1609. The endothelium-independent dilatation elicited by isoproterenol was not altered by either NO donor. In NG-nitro-L-arginine–treated carotid artery segments, C87-3786 significantly attenuated the acetylcholine-induced increase in 6-keto-prostaglandin F1
release, which was taken as an index of arachidonic acid liberation. In parallel experiments using cultured human endothelial cells, C87-3786 attenuated the Ca2+ response to bradykinin. The release of EDHF from a luminally perfused porcine coronary artery was detected by recording the membrane potential of downstream-situated cultured rat aortic smooth muscle cells. The NO donor C87-3786 had no effect on the hyperpolarization elicited by preformed EDHF but markedly inhibited its release from bradykinin-stimulated donor segments.
Conclusions These findings indicate that under physiological conditions, the production of EDHF is damped by NO. Therefore, it follows that when NO synthesis is impaired, alleviation of this intrinsic inhibition may, at least in part, maintain endothelial vasodilator function.
Key Words: endothelium-derived factors • vasodilation • calcium • acetylcholine • bradykinin
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Introduction |
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In addition to NO and PGI2, endothelium-dependent vasodilatation is mediated by the release of a third endothelium-derived factor, which causes hyperpolarization of the underlying smooth muscle cells (for review, see References 1 and 2). The chemical nature of this EDHF has been only partly characterized. However, in line with a previous hypothesis,3 4 5 several recent studies confirmed that in the coronary macrocirculation and microcirculation6 7 8 9 as well as in rabbit carotid arteries,10 a cytochrome P450–derived arachidonic acid metabolite displays the characteristics of EDHF.
The contribution of EDHF to endothelium-dependent vasodilatation under physiological conditions is difficult to assess, because EDHF-mediated dilatations can be discerned only after inhibition of NOS. Although treatment with specific NOS inhibitors at most slightly attenuates the maximal endothelium-dependent dilatations in the rat coronary microcirculation7 and in rat mesenteric,11 porcine coronary,6 12 and canine coronary13 14 arteries, these observations do not necessarily imply a major role for EDHF in endothelium-dependent vasodilatation in these vascular beds. In fact, although it has been shown that EDHF generates a maximal dilatation after NOS blockade, NO accounts almost entirely for agonist-induced dilatations in porcine epicardial arteries under physiological conditions.15 Thus, it is conceivable that NO exerts a feedback inhibition on EDHF formation. Therefore, we investigated the effects of NO donors on EDHF-mediated dilator responses in perfused segments of rabbit carotid and porcine coronary arteries as well as on EDHF release and EDHF-mediated hyperpolarization. In addition, the effect of an NO donor on bradykinin-stimulated Ca2+ signaling, an initiating event in EDHF generation,16 17 was investigated in cultured human endothelial cells.
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Methods |
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Materials
Acetylcholine, phenylephrine, isoproterenol, tetrabutylammonium, and nystatin were purchased from Sigma Chemical Co; bradykinin from Bachem Biochemica GmbH; HEPES and L-NNA from Serva; M-119 medium from GIBCO; penicillin, streptomycin, L-glutamine, glutathione, and L(+)-ascorbic acid (Biotect protection medium) from Biochrom; diclofenac (Voltaren injection solution) from CIBA-Geigy; fura 2-AM and ionomycin from Calbiochem-Novabiochem GmbH; pluronic F-127 from Molecular Probes; oxyhemoglobin from Calzyme Laboratories; and pentobarbital sodium (Nembutal) from Sanofi. U46619 (9,11-dideoxy-11
,9-epoxymethano-prostaglandin F2) was provided by Upjohn and C87-3786 (N-[cis-2,6-dimethylpiperidino]-N-nitroso-2-aminoacetonitril) and CAS 1609 (4-hydroxymethyl-furoxan-3-carboxamide) by Hoechst-Casella. NS 2028 (1H-[1,2,4]oxadiazolo-[4,3-a]-6-bromo-quinoxazin-1-one) was provided by NeuroSearch and superoxide dismutase (Peroxinorm) by Grunenthal GmbH. NO gas was prepared by the reaction of FeSO4 with sodium nitrite as described.18
Cell Culture
Endothelial cells
Human umbilical vein endothelial cells isolated from umbilical cords as described19 were seeded on quartz coverslips in culture dishes containing medium M-119 and 20% heat-inactivated FCS (Vitromex) supplemented with penicillin (50 U/mL), streptomycin (50 µg/mL), L-glutamine (1 mmol/L), glutathione (5 mg/mL), and L(+)-ascorbic acid (5 mg/mL). [Ca2+]i was estimated in cells grown on coverslips for 24 to 48 hours.
Smooth muscle cells
Rat aortic smooth muscle cells were isolated and cultured as described.20 Confluent cultures of smooth muscle cells were passaged by use of trypsin-EDTA (1 mmol/L), and experiments were performed with cells between passages 12 and 16.
Vessel Preparation
Rabbit carotid arteries
New Zealand White rabbits of either sex (1.5 to 2.5 kg body wt) were anesthetized with pentobarbital sodium (60 mg/kg IV) and exsanguinated by cuts through both the aorta and vena cava. Both carotid arteries were dissected, cleaned of adventitial adipose and connective tissue, and cut into segments 10 mm long for diameter registration and 25 mm long for determination of 6-keto-PGF1 release.
Porcine coronary arteries
Porcine hearts were obtained from a local slaughterhouse, placed immediately into ice-cold Krebs-Henseleit solution (see below), and transported to the laboratory. The coronary arteries were dissected, cleaned of adventitial adipose and connective tissue, and cut into segments 20 mm long for diameter registration and 50 mm long for the bioassay of EDHF.
Diameter Registration
Diameter registration was performed as described.21 Briefly, rabbit carotid artery segments were cannulated at both ends and placed into organ chambers containing Tyrode's solution of the following composition (mmol/L): Na+ 144.3, K+ 4.0, Cl- 138.6, Ca2+ 1.7, Mg2+ 1.0, HCO3- 11.9, and glucose 10.0 and containing the cyclooxygenase inhibitor diclofenac (1 µmol/L). The extraluminal solution was gassed with 95% O2/5% CO2 to give a PO2 of >300 mm Hg, and the luminal perfusate was gassed with 20% O2/5% CO2/75% N2 to give a PO2 of 140 mm Hg (37°C, pH 7.4). Perfusion routes for the chamber perfusion and the vessel lumen were separate, and drugs were administered to either route independently. The perfusion pressure was adjusted to 50 mm Hg, and vessels were gradually stretched to their in situ length during an initial equilibration period (60 minutes). Thereafter, the vessel lumen was perfused with Tyrode's solution (0.5 mL/min), and the outer vessel diameter was recorded continuously by a photoelectric device. Under these conditions, the carotid artery diameter was determined to be 1667±19 µm (n=68). The carotid arteries were then preconstricted with phenylephrine (1 to 3 µmol/L) to achieve similar constriction levels (292±17 µm) in either the presence or absence of L-NNA or of the NO donors.
In the case of porcine coronary arteries (resting diameter, 3560±375 µm [n=10]), preconstriction (643±89 µm) was achieved by the addition of the thromboxane mimetic U46619 (0.1 to 0.3 µmol/L) to the intraluminal perfusate.
Detection of EDHF Release
The release of EDHF was detected by recording changes in membrane potential of cultured rat aortic smooth muscle cells exposed to the effluate of a perfused porcine coronary artery. The transit time between the coronary artery and the detector smooth muscle cells was 
2 seconds. Briefly, a segment of porcine coronary artery was cannulated at both ends, mounted in an organ chamber, and perfused intraluminally (1 mL/min) with physiological salt solution of the following composition (in mmol/L): NaCl 140, KCl 4.7, MgCl2 1, CaCl2 1.3, glucose 5, and HEPES 10 (pH 7.4, 37°C) containing the NOS inhibitor L-NNA (100 µmol/L) and diclofenac (1 µmol/L). Effluate from the segment superfused the cultured smooth muscle cells, the membrane potential of which was recorded by the slow whole-cell configuration22 of the patch-clamp technique.23 The patch pipettes had an input resistance of 8 to 10 M
when filled with KCl solution containing (in mmol/L) KCl 140, MgCl2 1, CaCl2 1.3, HEPES 10, and glucose 5 (pH 7.4). Gigaohm seals were established by gentle suction. The slow whole-cell configuration was obtained with nystatin (100 µg/mL) in the pipette. This nystatin concentration provides a low-resistance access to the cytosol to measure intracellular potentials under current-clamp conditions. The electrical contact with the cytosol was established within 1 to 2 minutes of seal formation. The cell membrane potentials, measured in the current-clamp mode, were recorded continuously. Only detector smooth muscle cells that had a stable resting membrane potential for >2 minutes and exhibited no further change in the input resistance were used in the bioassay system.
Radioimmunoassay for 6-Keto-PGF1
In a separate series of experiments, the concentration of 6-keto-PGF1, the stable hydrolysis product of PGI2, in the effluate of perfused (0.166 mL/min) rabbit carotid arteries (collected for 6 minutes before and after application of acetylcholine 1 µmol/L in the presence of L-NNA 100 µmol/L) was measured by a specific radioimmunoassay.24
Measurement of [Ca2+]i
For the measurement of [Ca2+]i, endothelial cells were loaded with the fluorescent Ca2+-sensitive dye fura 2 by incubation with 3 µmol/L fura 2-AM and 0.025% (wt/vol) Pluronic F-127 at 37°C for 60 minutes. Thereafter, the coverslips were washed in HEPES-modified Tyrode's solution of the following composition (mmol/L): Na+ 136, K+ 4.0, Cl- 138.5, Ca2+ 1.0, Mg2+ 0.5, HEPES 9.5, and glucose 5. [Ca2+]i was determined fluorometrically in thermostatically controlled cuvettes as described previously.25
Statistics
Dilator responses are given as percentage dilatation relative to the preconstriction level. All data in the figures and text are expressed as mean±SEM of n experiments with segments from different arteries. Statistical analysis was performed by one-way ANOVA followed by a Bonferroni t test or by the Mann-Whitney test for unpaired data, where appropriate, with values of P<.05 considered statistically significant.
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Results |
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Effect of NO Donors on EDHF-Mediated Dilatation
In rabbit carotid artery segments preconstricted with phenylephrine (1 to 3 µmol/L), acetylcholine (0.03 to 3 µmol/L) elicited a concentration-dependent dilatation (Fig 1
). In the presence of the NOS inhibitor L-NNA (100 µmol/L), the concentration-response curve was displaced to the right (EC50 of 59 nmol/L versus 330 nmol/L), but the maximal dilatation remained virtually unchanged (Fig 1). However, the L-NNA/diclofenac–resistant dilatation was completely abolished after pretreatment with tetrabutylammonium (1 mmol/L), an inhibitor of Ca2+-activated K+ channels (Fig 1). Superoxide dismutase (30 nmol/L), on the other hand, failed to affect the L-NNA/diclofenac–resistant dilatation (18±2% versus 25±8% and 79±7% versus 85±7% dilatation at 0.1 µmol/L and 1 µmol/L acetylcholine, respectively, n=5).
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) and presence (
) of L-NNA (100 µmol/L) and in the presence of L-NNA and tetrabutylammonium (1 mmol/L, 
). Results are expressed as the mean±SEM of five separate experiments. *P<.05, **P<.01 vs respective control.This L-NNA/diclofenac–insensitive dilatation, which is generally attributed to EDHF, was investigated in the presence of two different NO donors, C87-3786 (3 µmol/L) and CAS 1609 (3 µmol/L),26 27 which were applied in a concentration that in another experimental series abrogated the supplemental L-NNA–induced constriction (253±28 µm for phenylephrine 1 µmol/L alone, 346±29 µm for phenylephrine in the presence of L-NNA, and 264±15 µm for phenylephrine in the presence of L-NNA and the NO donors, n=5 to 10).
C87-3786 and CAS 1609 significantly attenuated the acetylcholine-induced, EDHF-mediated dilatations attributed to the release of EDHF (Fig 2A and 2B). In contrast, in the absence of L-NNA, C87-3786 (10 µmol/L) had no effect on the presumably NO-mediated vasodilatation elicited by acetylcholine (Fig 2C). Similarly, the endothelium-independent vasodilatation induced by isoproterenol was preserved in the presence of the NO donor (39±6% versus 43±8% dilatation and 59±8% versus 64±3% dilatation at 0.1 µmol/L and 10 µmol/L isoproterenol, respectively, n=4).
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) or (B) CAS1609 (3 µmol/L, 
) did not affect the acetylcholine-induced dilatation (Pretreatment with the potent competitive inhibitor of soluble guanylyl cyclase NS 2028 (1 µmol/L), which completely abolished the dilator response to C87-3786 and the associated increase in vascular cGMP concentration (from 2.1±0.4 to 0.37±0.07 pmol/mg protein, n=5, P<.05, unpublished observation), affected neither the L-NNA/diclofenac–resistant dilatation nor the inhibitory effect of the NO donor C87-3786 (NS 2028, 22±3% versus NS 2028 plus C87-3786, 10±2% dilatation at 0.1 µmol/L acetylcholine and 82±6% versus 49±4% dilatation at 1 µmol/L acetylcholine, n=5).
In perfused segments of porcine epicardial arteries preconstricted with U46619 (0.1 to 0.3 µmol/L) and pretreated with L-NNA (100 µmol/L), addition of bradykinin (1 to 100 nmol/L) to the luminal perfusate elicited a concentration-dependent dilatation. Pretreatment with C87-3786 (3 µmol/L) significantly attenuated the bradykinin-induced dilator response (Fig 3).
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Effect of C87-3786 on EDHF Release From Porcine Coronary Arteries and EDHF-Mediated Hyperpolarizations
To elucidate the interaction between NO and EDHF, the effects of C87-3786 were analyzed in a patch-clamp system for the detection of EDHF. In this system, the interference of NO with EDHF release from the donor endothelium could be investigated separately from the effects of NO on the membrane potential and EDHF-induced hyperpolarization of the detector cells (Fig 4).
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In the presence of both diclofenac and L-NNA, stimulation of the donor porcine epicardial artery with bradykinin (100 nmol/L) led to a hyperpolarization of the detector smooth muscle cells of 13.7±1.6 mV. Pretreatment of the detector cells with tetrabutylammonium (1 mmol/L) abrogated this hyperpolarization (R.P. et al, unpublished observations, 1996). Addition of C87-3786 (10 or 100 µmol/L) to the perfusate significantly reduced the EDHF-mediated hyperpolarizations (Fig 4
). In contrast, direct application of the NO donor to the detector smooth muscle cells did not affect EDHF-mediated hyperpolarizations after bradykinin stimulation (Fig 4). Comparable results were obtained with cultured porcine coronary artery smooth muscle cells.
Pretreatment with the inhibitor of soluble guanylyl cyclase NS 2028 (1 µmol/L) failed to affect EDHF-mediated hyperpolarizations and the inhibitory effect of C87-3786 on EDHF-mediated hyperpolarizations (Fig 4B).
Neither superoxide dismutase (100 nmol/L) nor oxyhemoglobin (0.1 µmol/L) had any effect on the duration or the amplitude of the EDHF-mediated hyperpolarizations (-12.8±2 and -11.2±1.5 mV, respectively, n=6). On the other hand, oxyhemoglobin strongly attenuated the hyperpolarizing effect of NO applied as a bolus (1 nmol) to the superfusate proximal to the detector cells (from -7.2±1.1 to -1.6±1.2 mV, n=4, P<.05).
No differences in resting membrane potential were observed with the various treatments (n=6 for each treatment): control, -43±2 mV; C87-3786 (10 µmol/L, distal application), -44±4 mV; C87-3786 (10 µmol/L, proximal application), -44±3 mV; C87-3786 (100 µmol/L, proximal application), -43±4 mV; NS 2028 (1 µmol/L, proximal application), -42±3 mV; NS 2028 and C87-3786 (1 µmol/L and 10 µmol/L, proximal application), -43±3 mV; superoxide dismutase (100 µmol/L, proximal application), -45±2 mV; and oxyhemoglobin (0.1 µmol/L, proximal application), -43±3 mV.
Modulation of Ca2+ Signaling by C87-3786
In human umbilical vein endothelial cells pretreated with L-NNA (100 µmol/L), stimulation with bradykinin (10 nmol/L) induced a transient peak increase in [Ca2+]i from 109±8 to 418±35 nmol/L (n=9). Pretreatment with C87-3786 (100 µmol/L, 15 minutes) significantly attenuated the peak bradykinin-induced [Ca2+]i increase without affecting basal values (Fig 5).
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Effect of C87-3786 on 6-Keto-PGF1 Release From Rabbit Carotid Arteries
The liberation of arachidonic acid, the putative precursor of EDHF, was assessed by the accumulation of the stable arachidonic acid derivative 6-keto-PGF1 in the effluate from carotid artery segments not exposed to diclofenac. In the presence of L-NNA (100 µmol/L), perfusion with acetylcholine (1 µmol/L) elicited an increase in 6-keto-PGF1 release from 92±8 to 298±85 pg/mL (n=5). The acetylcholine-induced increase in 6-keto-PGF1 was significantly attenuated in the presence of C87-3786 (10 µmol/L) (Fig 6).
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Discussion |
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The results of the present study demonstrate that exogenously applied NO attenuates EDHF-mediated dilatations in rabbit carotid and porcine coronary arteries. This inhibitory effect resulted from interference with the synthesis and/or release of EDHF rather than its mechanism of action. Indeed, patch-clamp recordings in cultured vascular smooth muscle cells revealed that NO donors, which directly affected neither resting membrane potential nor EDHF-induced hyperpolarization, markedly attenuated EDHF release from a donor segment.
Although EDHF-mediated dilatation has been described in various vascular beds, it appears to play a particularly significant role in the coronary macrocirculation and microcirculation.7 12 28 However, such a physiological function has been attributed to EDHF mainly by analogy in experiments in which the production of other endothelium-dependent autacoids was inhibited. Therefore, it is unclear whether EDHF is a constitutive component of the agonist-induced endothelium-dependent dilatation or is synthesized only after suppression of NO and PGI2 release. In the latter case, exogenous application of NO would be expected to suppress EDHF-mediated dilatations. Our results substantiate this hypothesis in that the NO donors C87-3786 and CAS 1609 attenuated the agonist-induced dilator response attributed to EDHF in rabbit carotid and porcine coronary arteries under conditions of combined NOS and cyclooxygenase blockade. The NO donors used throughout the present study were administered at a concentration that just reversed the L-NNA–induced constriction. We thereby minimized the risk of detecting nonspecific effects related to excessively high concentrations of NO by applying physiologically relevant NO levels.
In designing the patch-clamp experiments, we took into account the possibility that NO may be able to interact with EDHF at the level of its generation as well as at its site of action. With regard to the interference at the site of action, EDHF-induced hyperpolarization is thought to be mediated by the opening of Ca2+-activated K+ channels.6 12 Although NO has been reported to directly activate Ca2+-activated K+ channels in rabbit aortic smooth muscle cells29 and to induce hyperpolarization in coronary arteries under certain experimental conditions,30 these findings are somewhat controversial.31 32 33 In the patch-clamp system for the detection of EDHF, the experiments using oxyhemoglobin as a scavenger for NO revealed that in the presence of L-NNA, NO synthesis was effectively abrogated. Together with the lack of effect of superoxide dismutase, these results can be taken as evidence that, as reported for bovine coronary arteries,34 the L-NNA–resistant response was not mediated by NO formed either by NOS or by any other pathway. The direct application of the NO donor C87-3786 to the smooth muscle cells did not alter resting membrane potential, indicating that at this concentration of NO, the results obtained were not complicated by a direct effect on the detector cells. Thus, the marked reduction in EDHF-mediated hyperpolarization observed after inclusion of C87-3786 in the donor perfusate suggests that NO inhibits the synthesis and/or release of EDHF from endothelial cells.
EDHF formation in response to receptor-dependent agonists appears to be strictly dependent on an increase in the intracellular concentration of Ca2+16 17 and is likely to involve the formation of a Ca2+-calmodulin complex.35 Moreover, a close association between the increase in [Ca2+]i, arachidonic acid release, and EDHF-mediated relaxations in porcine coronary arteries has recently been described.36
A negative feedback inhibition of EDHF production by NO may involve alterations in Ca2+ signaling in endothelial cells, since inhibition of NOS enhances the Ca2+ response to receptor-dependent agonists.37 The observed attenuation of the Ca2+ response to bradykinin by the NO donor C87-3786 in human endothelial cells pretreated with L-NNA therefore provides an explanation for the reduction of EDHF formation by NO. This hypothesis is further supported by the finding that C87-3786 reduces the release of 6-keto-PGF1 from native endothelial cells. Since phospholipase A2, the rate-limiting enzyme for the liberation of arachidonic acid from phospholipids, is the only strictly Ca2+-dependent enzyme in the biosynthetic cascade of 6-keto-PGF1, it is conceivable that the NO-mediated decrease in [Ca2+]i results in an inhibition of EDHF formation.
Although it could be expected that an NO-induced decrease in [Ca2+]i would affect NO synthesis, it should be borne in mind that the effect on [Ca2+]i is relatively small and is more likely to result in a marked inhibition of an enzyme with a low Ca2+ sensitivity (eg, phospholipase A2) than the highly Ca2+-sensitive NOS. This supposition is reinforced by the fact that in endothelial cells the [Ca2+]i threshold for prostacyclin formation is markedly greater than that required for NO synthesis.38 The mechanisms by which NO is able to attenuate endothelial Ca2+ signaling and EDHF release remain unclear; however, the results obtained with the specific guanylyl cyclase inhibitor NS 2028 indicate that NO exerts its effects independently of an increase in vascular cGMP concentration. This is consistent with a cGMP-independent effect of NO on [Ca2+]i in fibroblasts.39
Besides its effects on [Ca2+]i, NO may attenuate the formation of EDHF by interfering with the EDHF-generating enzyme(s). Indeed, inhibition of hepatic microsomal P450 by NO has been attributed to an interaction with the heme domain.40 However, inhibition of P450 enzymes required relatively high concentrations of NO, ie, levels similar to that produced by the inducible, Ca2+-independent NOS. In endothelial cells pretreated with the P450-inducing compound ß-naphthoflavone, P450 activity was attenuated by NO donors (I.F. and A. Mulsch, PhD, unpublished observations, 1996).
We observed a strong attenuating effect of a relatively low concentration of an NO donor on arachidonic acid liberation, as indexed by a reduction in 6-keto-PGF1 formation in rabbit carotid arteries. This observation suggests that NO may act at a different point of the signal transduction cascade, namely, the liberation of the putative substrate for "EDHF synthase."
The observed attenuation of EDHF-mediated dilatation by NO may imply that alleviation of this intrinsic inhibitory pathway under conditions of impaired NO release leads to an enhanced formation of EDHF. Therefore, in pathophysiological states such as hypercholesterolemia, hypertension, arteriosclerosis, and diabetes, which are associated with decreased bioavailability of endothelium-derived NO,41 EDHF formation may be of much greater importance than under physiological conditions. Indeed, in a rabbit model of hypercholesterolemia, endothelium-dependent hyperpolarization, mediated via opening of Ca2+-activated K+ channels, maintains endothelium-dependent dilatation in response to acetylcholine.42 Moreover, in the same model, an enhanced synthesis of cytochrome P450–derived epoxyeicosatrienoic acids, which activate Ca2+-activated K+ channels,43 has been described.44
In conclusion, our data indicate that physiologically relevant concentrations of NO attenuate EDHF-mediated dilatations via a decreased formation of this hyperpolarizing factor. Under conditions of impaired NO synthesis, however, EDHF formation is unimpeded and may maintain endothelial vasodilator function.
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Selected Abbreviations and Acronyms |
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Acknowledgments |
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This work was supported by the Deutsche Forschungsgemeinschaft (Bu 436/4-3, He 1587/5-1). The authors wish to thank Isabel Winter, Michaela Staechele, and Edeltraut Thielen for expert technical assistance.
Received June 27, 1996; revision received July 29, 1996; accepted August 7, 1996.
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References |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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1. Garland CJ, Plane F, Kemp BK, Cocks TM. Endothelium-dependent hyperpolarization: a role in the control of vascular tone. Trends Pharmacol Sci. 1995;16:23-30.[Medline] [Order article via Infotrieve]
2.
Cohen RA, Vanhoutte PM. Endothelium-dependent hyperpolarization: beyond nitric oxide and cyclic GMP. Circulation. 1995;92:3337-3349.
3. Rubanyi GM, Vanhoutte PM. Nature of endothelium-derived relaxing factor: are there two relaxing mediators? Circ Res. 1987;61:61-67.
4.
Rubanyi GM, McKinney M, Vanhoutte PM. Biphasic release of endothelium-derived relaxing factor(s) by acetylcholine from perfused canine femoral arteries: characterization of muscarinic receptors. J Pharmacol Exp Ther. 1987;240:802-808.
5. Komori K, Vanhoutte PM. Endothelium-derived hyperpolarizing factor. Blood Vessels. 1990;27:238-245.[Medline] [Order article via Infotrieve]
6. Hecker M, Bara A, Bauersachs J, Busse R. Characterization of endothelium-derived hyperpolarizing factor as a cytochrome P450-derived arachidonic acid metabolite in mammals. J Physiol (Lond). 1994;4812:407-414.
7. Bauersachs J, Hecker M, Busse R. Display of the characteristics of endothelium-derived hyperpolarizing factor by a cytochrome P450-derived arachidonic acid metabolite in the coronary microcirculation. Br J Pharmacol. 1994;113:1548-1553.[Medline] [Order article via Infotrieve]
8. Fulton D, McGiff JC, Quilley J. Role of K+ channels in the vasodilator response to bradykinin in the rat heart. Br J Pharmacol. 1994;113:954-958.[Medline] [Order article via Infotrieve]
9. Fulton D, Mahboubi K, McGiff JC, Quilley J. Cytochrome P450-dependent effects of bradykinin in the rat heart. Br J Pharmacol. 1995;114:99-102.[Medline] [Order article via Infotrieve]
10. Lischke V, Busse R, Hecker M. Selective inhibition by barbiturates of the synthesis of endothelium-derived hyperpolarizing factor in the rabbit carotid artery. Br J Pharmacol. 1995;115:969-974.[Medline] [Order article via Infotrieve]
11.
Nagao T, Illiano S, Vanhoutte PM. Heterogenous distribution of endothelium-dependent relaxations resistant to NG-nitro-L-arginine in rats. Am J Physiol. 1992;263:H1090-H1094.
12.
Nagao T, Vanhoutte PM. Hyperpolarization as a mechanism for endothelium-dependent relaxation in the porcine coronary artery. J Physiol (Lond). 1992;445:355-367.
13.
Mombouli J-V, Illiano S, Nagao T, Scott-Burden T, Vanhoutte PM. Potentiation of endothelium-dependent relaxations to bradykinin by angiotensin I converting enzyme inhibitors in canine coronary artery involves both endothelium-derived relaxing and hyperpolarizing factors. Circ Res. 1992;71:137-144.
14. Mombouli J-V, Nakashima M, Hamra M, Vanhoutte PM. Endothelium-dependent relaxation and hyperpolarization evoked by bradykinin in canine coronary arteries: enhancement by exercise-training. Br J Pharmacol. 1996;117:413-418.[Medline] [Order article via Infotrieve]
15. Kilpatrick EV, Cocks TM. Evidence for differential roles of nitric oxide (NO) and hyperpolarization in endothelium-dependent relaxation of pig isolated coronary artery. Br J Pharmacol. 1994;112:557-565.[Medline] [Order article via Infotrieve]
16.
Busse R, Fichtner H, Luckhoff A, Kohlhardt M. Hyperpolarization and increased free calcium in acetylcholine-stimulated endothelial cells. Am J Physiol. 1988;255:H965-H969.
17.
Chen G, Suzuki H. Calcium dependency of the endothelium-dependent hyperpolarization in smooth muscle cells of the rabbit carotid artery. J Physiol (Lond). 1990;421:521-534.
18. Vedernikov JP, Graser T, Vanin AF. Similar endothelium-independent arterial relaxation by carbon monoxide and nitric oxide. Biomed Biochim Acta. 1989;48:595-599.
19. Busse R, Lamontagne D. Endothelium-derived bradykinin is responsible for the increase in calcium produced by angiotensin-converting enzyme inhibitors in human endothelial cells. Naunyn Schmiedebergs Arch Pharmacol. 1991;344:126-129.[Medline] [Order article via Infotrieve]
20.
Schini VB, Catovsky B, Schray-Utz B, Busse R, Vanhoutte PM. Insulin-like growth factor I inhibits induction of nitric oxide synthase in vascular smooth muscle cells. Circ Res. 1994;74:24-32.
21. Busse R, Pohl U, Kellner C, Klemm U. Endothelial cells are involved in the vasodilatory response to hypoxia. Pflugers Arch. 1983;397:78-80.[Medline] [Order article via Infotrieve]
22.
Horn R, Marty A. Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol. 1988;92:145-159.
23. Hamill OP, Marty A, Neher E, Sakman B, Sigworth FJ. Improved patch clamp techniques for high resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981;391:85-100.[Medline] [Order article via Infotrieve]
24. Lamontagne D, Konig A, Bassenge E, Busse R. Prostacyclin and nitric oxide contribute to the vasodilator action of acetylcholine and bradykinin in the intact rabbit coronary bed. J Cardiovasc Pharmacol. 1992;20:652-657.[Medline] [Order article via Infotrieve]
25.
Fleming I, Hecker M, Busse R. Intracellular alkalinization induced by bradykinin sustains activation of the constitutive nitric oxide synthase in endothelial cells. Circ Res. 1994;74:1220-1226.
26. Mulsch A, Hecker M, Mordvintcev PI, Vanin AF, Busse R. Enzymic and nonenzymic release of NO accounts for the vasodilator activity of the metabolites of CAS 936, a novel long-acting sydnonimine derivative. Naunyn Schmiedebergs Arch Pharmacol. 1993;347:92-100.[Medline] [Order article via Infotrieve]
27. Hecker M, Vorhoff W, Bara A, Mordvintcev PI, Busse R. Characterization of furoxans as a new class of tolerance-resistant nitrovasodilators. Naunyn Schmiedebergs Arch Pharmacol. 1995;351:426-432.[Medline] [Order article via Infotrieve]
28. Nakashima M, Mombouli J-V, Taylor AA, Vanhoutte PM. Endothelium-dependent hyperpolarization caused by bradykinin in human coronary arteries. J Clin Invest. 1993;92:2867-2871.
29. Bolotina VM, Najibi S, Palacino JJ, Pagano PJ, Cohen RA. Nitric oxide directly activates calcium-dependent potassium channels in vascular smooth muscle. Nature. 1994;368:850-853.[Medline] [Order article via Infotrieve]
30.
Parkington HC, Tare M, Tonta MA, Coleman HA. Stretch revealed three components in the hyperpolarization of guinea-pig coronary artery in response to acetylcholine. J Physiol (Lond). 1993;465:459-476.
31. Taylor SG, Southerton JS, Weston AH, Baker JRJ. Endothelium-dependent effects of acetylcholine in rat aorta: a comparison with sodium nitroprusside and cromakalin. Br J Pharmacol. 1988;94:853-863.[Medline] [Order article via Infotrieve]
32. Rand VE, Garland CJ. Endothelium-dependent relaxation to acetylcholine in the rabbit basilar artery: importance of membrane hyperpolarization. Br J Pharmacol. 1992;106:143-150.[Medline] [Order article via Infotrieve]
33. Komori K, Lorenz RR, Vanhoutte PM. Nitric oxide, ACh, and electrical and mechanical properties of canine arterial smooth muscle. Am J Physiol. 1988;255:143-150.
34. Drummond GR, Cocks TM. Evidence for mediation by endothelium-derived hyperpolarizing factor of relaxation to bradykinin in the bovine isolated coronary independently of voltage-operated Ca2+ channels. Br J Pharmacol. 1996;117:1035-1040.[Medline] [Order article via Infotrieve]
35. Illiano S, Nagao T, Vanhoutte PM. Calmidazolium, a calmodulin inhibitor, inhibits endothelium-dependent relaxations resistant to nitro-L-arginine in the canine coronary artery. Br J Pharmacol. 1992;107:387-392.[Medline] [Order article via Infotrieve]
36.
Weintraub NL, Stephenson AH, Sprague RS, McMurdo L, Lonigro AJ. Relationship of arachidonic acid release to porcine coronary artery relaxation. Hypertension. 1995;26:684-690.
37.
Shin WS, Sasaki T, Kato M, Hara K, Seko A, Yang W-D, Shimamoto N, Sugimoto T, Toyo-oka T. Autocrine and paracrine effects of endothelium-derived relaxing factor on intracellular Ca2+ of endothelial cells and vascular smooth muscle cells. J Biol Chem. 1992;267:20377-20382.
38. Parsaee H, McEwan JR, Joseph S, MacDermot J. Differential sensitivities of the prostacyclin and nitric oxide biosynthetic pathway to cytosolic calcium in bovine aortic endothelial cells. Br J Pharmacol. 1992;107:1013-1019.[Medline] [Order article via Infotrieve]
39.
Garg CG, Hassid A. Nitric oxide decreases cytosolic free calcium in Balb/c 3T3 fibroblasts by a cyclic GMP-independent mechanism. J Biol Chem. 1991;266:9-12.
40.
Khatsenko OG, Gross SS, Rifkind AB, Vane JR. Nitric oxide is a mediator of the decrease in cytochrome P450-dependent metabolism caused by immunostimulants. Proc Natl Acad Sci U S A. 1993;90:11147-11151.
41. Busse R, Fleming I. Endothelial dysfunction in atherosclerosis. J Vasc Res. 1996;33:181-194.[Medline] [Order article via Infotrieve]
42.
Najibi S, Cowan CL, Palacino JJ, Cohen RA. Enhanced role of potassium channels in relaxations to acetylcholine in hypercholesterolemic rabbit carotid artery. Am J Physiol. 1994;266:H2061-H2067.
43. Hu S, Kim HS. Activation of K+ channel in vascular smooth muscles by cytochrome P450 metabolites of arachidonic acids. Eur J Pharmacol. 1993;230:215-221.[Medline] [Order article via Infotrieve]
44.
Pfister SL, Falck JR, Campbell WB. Enhanced synthesis of epoxyeicosatrienoic acids by cholesterol-fed rabbit aorta. Am J Physiol. 1991;261:H843-H852.
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|
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|
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|
|
|
|
|
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|
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|
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|
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|
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A. Huang, Y. Wu, D. Sun, A. Koller, and G. Kaley Effect of estrogen on flow-induced dilation in NO deficiency: role of prostaglandins and EDHF J Appl Physiol, December 1, 2001; 91(6): 2561 - 2566. [Abstract] [Full Text] [PDF]
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J. H Traverse, Y. Chen, M. Crampton, S. Voss, and R. J Bache Increased extravascular forces limit endothelium-dependent and -independent coronary vasodilation in congestive heart failure Cardiovasc Res, December 1, 2001; 52(3): 454 - 461. [Abstract] [Full Text] [PDF]
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H. L. Xu, R. A. Santizo, H. M. Koenig, and D. A. Pelligrino Chronic estrogen depletion alters adenosine diphosphate-induced pial arteriolar dilation in female rats Am J Physiol Heart Circ Physiol, November 1, 2001; 281(5): H2105 - H2112. [Abstract] [Full Text] [PDF]
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I. Fleming Cytochrome P450 and Vascular Homeostasis Circ. Res., October 26, 2001; 89(9): 753 - 762. [Abstract] [Full Text] [PDF]
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W. M. Chilian and R. Koshida EDHF and NO: Different Pathways for Production--Similar Actions Circ. Res., October 12, 2001; 89(8): 648 - 649. [Full Text] [PDF]
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C. L. Oltman, N. L. Kane, J. L. Fudge, N. L. Weintraub, and K. C. Dellsperger Endothelium-derived hyperpolarizing factor in coronary microcirculation: responses to arachidonic acid Am J Physiol Heart Circ Physiol, October 1, 2001; 281(4): H1553 - H1560. [Abstract] [Full Text] [PDF]
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S. P. Marrelli Mechanisms of endothelial P2Y1- and P2Y2-mediated vasodilatation involve differential [Ca2+]i responses Am J Physiol Heart Circ Physiol, October 1, 2001; 281(4): H1759 - H1766. [Abstract] [Full Text] [PDF]
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G.-W. He and Z.-G. Liu Comparison of Nitric Oxide Release and Endothelium-Derived Hyperpolarizing Factor-Mediated Hyperpolarization Between Human Radial and Internal Mammary Arteries Circulation, September 18, 2001; 104 (2009): I-344 - I-349. [Abstract] [Full Text] [PDF]
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W. B. Campbell and D. R. Harder Prologue: EDHF-what is it? Am J Physiol Heart Circ Physiol, June 1, 2001; 280(6): H2413 - H2416. [Full Text] [PDF]
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Y. Wu, A. Huang, D. Sun, J. R. Falck, A. Koller, and G. Kaley Gender-specific compensation for the lack of NO in the mediation of flow-induced arteriolar dilation Am J Physiol Heart Circ Physiol, June 1, 2001; 280(6): H2456 - H2461. [Abstract] [Full Text] [PDF]
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A. Huang, D. Sun, M. A. Carroll, H. Jiang, C. J. Smith, J. A. Connetta, J. R. Falck, E. G. Shesely, A. Koller, and G. Kaley EDHF mediates flow-induced dilation in skeletal muscle arterioles of female eNOS-KO mice Am J Physiol Heart Circ Physiol, June 1, 2001; 280(6): H2462 - H2469. [Abstract] [Full Text] [PDF]
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J. P. J. Halcox, S. Narayanan, L. Cramer-Joyce, R. Mincemoyer, and A. A. Quyyumi Characterization of endothelium-derived hyperpolarizing factor in the human forearm microcirculation Am J Physiol Heart Circ Physiol, June 1, 2001; 280(6): H2470 - H2477. [Abstract] [Full Text] [PDF]
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R. Rastaldo, N. Paolocci, A. Chiribiri, C. Penna, D. Gattullo, and P. Pagliaro Cytochrome P-450 metabolite of arachidonic acid mediates bradykinin-induced negative inotropic effect Am J Physiol Heart Circ Physiol, June 1, 2001; 280(6): H2823 - H2832. [Abstract] [Full Text] [PDF]
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M. L. H. Honing, P. Smits, P. J. Morrison, J. C. Burnett Jr, and T. J. Rabelink C-Type Natriuretic Peptide-Induced Vasodilation Is Dependent On Hyperpolarization in Human Forearm Resistance Vessels Hypertension, April 1, 2001; 37(4): 1179 - 1183. [Abstract] [Full Text] [PDF]
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I. Fleming, U. R. Michaelis, D. Bredenkotter, B. Fisslthaler, F. Dehghani, R. P. Brandes, and R. Busse Endothelium-Derived Hyperpolarizing Factor Synthase (Cytochrome P450 2C9) Is a Functionally Significant Source of Reactive Oxygen Species in Coronary Arteries Circ. Res., January 19, 2001; 88(1): 44 - 51. [Abstract] [Full Text] [PDF]
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C. G. Sobey Potassium Channel Function in Vascular Disease Arterioscler Thromb Vasc Biol, January 1, 2001; 21(1): 28 - 38. [Abstract] [Full Text] [PDF]
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S. G. Clark and L. C. Fuchs BKCa channels compensate for loss of NOS-dependent coronary artery relaxation in cardiomyopathy Am J Physiol Heart Circ Physiol, December 1, 2000; 279(6): H2598 - H2603. [Abstract] [Full Text] [PDF]
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R. W. Jeremy and H. McCarron Effect of hypercholesterolemia on Ca2+-dependent K+ channel-mediated vasodilatation in vivo Am J Physiol Heart Circ Physiol, October 1, 2000; 279(4): H1600 - H1608. [Abstract] [Full Text] [PDF]
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K. Goto, K. Fujii, U. Onaka, I. Abe, and M. Fujishima Angiotensin-Converting Enzyme Inhibitor Prevents Age-Related Endothelial Dysfunction Hypertension, October 1, 2000; 36(4): 581 - 587. [Abstract] [Full Text] [PDF]
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 R. P. Brandes, F.-H. Schmitz-Winnenthal, M. Feletou, A. Godecke, P. L. Huang, P. M. Vanhoutte, I. Fleming, and R. Busse An endothelium-derived hyperpolarizing factor distinct from NO and prostacyclin is a major endothelium-dependent vasodilator in resistance vessels of wild-type and endothelial NO synthase knockout mice PNAS, August 15, 2000; 97(17): 9747 - 9752. [Abstract] [Full Text] [PDF]
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Y. Nishikawa, D. W. Stepp, and W. M. Chilian Nitric oxide exerts feedback inhibition on EDHF-induced coronary arteriolar dilation in vivo Am J Physiol Heart Circ Physiol, August 1, 2000; 279(2): H459 - H465. [Abstract] [Full Text] [PDF]
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E. Thorin, M. Lucas, P. Cernacek, and J. Dupuis Role of ETA receptors in the regulation of vascular reactivity in rats with congestive heart failure Am J Physiol Heart Circ Physiol, August 1, 2000; 279(2): H844 - H851. [Abstract] [Full Text] [PDF]
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B. Fisslthaler, N. Hinsch, T. Chataigneau, R. Popp, L. Kiss, R. Busse, and I. Fleming Nifedipine Increases Cytochrome P4502C Expression and Endothelium-Derived Hyperpolarizing Factor-Mediated Responses in Coronary Arteries Hypertension, August 1, 2000; 36(2): 270 - 275. [Abstract] [Full Text] [PDF]
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B. Vanheel and J. Van de Voorde EDHF and residual NO: different factors Cardiovasc Res, June 1, 2000; 46(3): 370 - 375. [Full Text] [PDF]
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E. Thorin, D. Meerkin, O. F. Bertrand, P. Paiement, M. Joyal, and R. Bonan Influence of Postangioplasty {beta}-Irradiation on Endothelial Function in Porcine Coronary Arteries Circulation, March 28, 2000; 101(12): 1430 - 1435. [Abstract] [Full Text] [PDF]
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F. H. de Man, A. W. E. Weverling-Rijnsburger, A. van der Laarse, A. H. M. Smelt, J. W. Jukema, and G. J. Blauw Not Acute but Chronic Hypertriglyceridemia Is Associated With Impaired Endothelium-Dependent Vasodilation : Reversal After Lipid-Lowering Therapy by Atorvastatin Arterioscler Thromb Vasc Biol, March 1, 2000; 20(3): 744 - 750. [Abstract] [Full Text] [PDF]
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A. Huang, D. Sun, C. J. Smith, J. A. Connetta, E. G. Shesely, A. Koller, and G. Kaley In eNOS knockout mice skeletal muscle arteriolar dilation to acetylcholine is mediated by EDHF Am J Physiol Heart Circ Physiol, March 1, 2000; 278(3): H762 - H768. [Abstract] [Full Text] [PDF]
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D. A. Pelligrino, R. A. Santizo, and Q. Wang Miconazole represses CO2-induced pial arteriolar dilation only under selected circumstances Am J Physiol Heart Circ Physiol, October 1, 1999; 277(4): H1484 - H1490. [Abstract] [Full Text] [PDF]
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S. Taddei, L. Ghiadoni, A. Virdis, S. Buralli, and A. Salvetti Vasodilation to Bradykinin Is Mediated by an Ouabain-Sensitive Pathway as a Compensatory Mechanism for Impaired Nitric Oxide Availability in Essential Hypertensive Patients Circulation, September 28, 1999; 100(13): 1400 - 1405. [Abstract] [Full Text] [PDF]
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Y. Nishikawa, D. W. Stepp, and W. M. Chilian In vivo location and mechanism of EDHF-mediated vasodilation in canine coronary microcirculation Am J Physiol Heart Circ Physiol, September 1, 1999; 277(3): H1252 - H1259. [Abstract] [Full Text] [PDF]
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D. Sun, A. Huang, C. J. Smith, C. J. Stackpole, J. A. Connetta, E. G. Shesely, A. Koller, and G. Kaley Enhanced Release of Prostaglandins Contributes to Flow-Induced Arteriolar Dilation in eNOS Knockout Mice Circ. Res., August 6, 1999; 85(3): 288 - 293. [Abstract] [Full Text] [PDF]
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J. E. da Silva-Santos and J. Assreuy Long-Lasting Changes of Rat Blood Pressure to Vasoconstrictors and Vasodilators Induced by Nitric Oxide Donor Infusion: Involvement of Potassium Channels J. Pharmacol. Exp. Ther., July 1, 1999; 290(1): 380 - 387. [Abstract] [Full Text]
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A. S Izzard and A. M Heagerty Impaired flow-dependent dilatation in distal mesenteric arteries from the spontaneously hypertensive rat J. Physiol., July 1, 1999; 518(1): 239 - 245. [Abstract] [Full Text] [PDF]
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M. Malmsjo, A. Bergdahl, X.-H. Zhao, X.-Y. Sun, T. Hedner, L. Edvinsson, and D. Erlinge Enhanced acetylcholine and P2Y-receptor stimulated vascular EDHF-dilatation in congestive heart failure Cardiovasc Res, July 1, 1999; 43(1): 200 - 209. [Abstract] [Full Text] [PDF]
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J. Kalliovalkama, P. Jolma, J.-P. Tolvanen, M. Kahonen, N. Hutri-Kahonen, X. Wu, P. Holm, and I. Porsti Arterial function in nitric oxide-deficient hypertension: influence of long-term angiotensin II receptor antagonism Cardiovasc Res, June 1, 1999; 42(3): 773 - 782. [Abstract] [Full Text] [PDF]
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K. Kusterer, T. Pohl, H.-P. Fortmeyer, W. Marz, H. Scharnagl, A. Oldenburg, S. Angermuller, I. Fleming, K. H. Usadel, and R. Busse Chronic selective hypertriglyceridemia impairs endothelium-dependent vasodilatation in rats Cardiovasc Res, June 1, 1999; 42(3): 783 - 793. [Abstract] [Full Text] [PDF]
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