Acetylcholine-mediated vasodilation in the forearm circulation of patients with heart failure: indirect evidence for the role of endothelium-derived hyperpolarizing factor

doi:10.1016/S0002-9149(01)01466-7

The American Journal of Cardiology

Volume 87, Issue 9, 1 May 2001, Pages 1089-1092

https://doi.org/10.1016/S0002-9149(01)01466-7 Get rights and content

Abstract

Vasomotor responses to intraarterial administration of acetylcholine are mediated by release of nitric oxide, prostaglandins, and an unidentified hyperpolarizing factor from vascular endothelial cells. The contribution of endothelium-derived hyperpolarizing factor (EDHF) to the vasodilatory response to acetylcholine in the skeletal muscle circulation of patients with congestive heart failure (CHF) has not been previously characterized. Accordingly, to specifically assess the role of EDHF, the regional vascular effects of sequential administration of acetylcholine and nitroglycerin in the brachial artery were determined in the forearm circulation with strain-gauge venous occlusion plethysmography in patients with CHF and in normal subjects during combined systemic inhibition of cyclooxygenase activity with indomethacin and regional inhibition of nitric oxide synthase activity with $l$ -N^G-monomethylarginine ( $l$ -NMMA). After administration of indomethacin, infusion of $l$ -NMMA significantly decreased the forearm blood flow response to acetylcholine in normal subjects (5.4 ± 1.2 to 3.5 ± 0.6 ml/min/100 ml, p < 0.05) but not in patients with CHF (5.7 ± 1.3 to 5.7 ± 1.4 ml/min/100 ml). Infusion of $l$ -NMMA did not change forearm blood flow responses to nitroglycerin in either group. The presence of a noncyclooxygenase, non-nitric-oxide relaxing factor indicates that EDHF, rather than nitric oxide, may be the predominant endothelium-derived substance mediating vasodilation in response to acetylcholine in patients with CHF.

Section snippets

Study group

Thirteen men and 1 woman (mean age, 50 ± 12 years [range 32 to 65]) with chronic stable CHF due to nonischemic cardiomyopathy were studied. The mean left ventricular ejection fraction was 22 ± 4%. Nine patients were in New York Heart Association functional class II and 5 patients were in functional class III. Cardiovascular medications, which included diuretics, angiotensin-converting enzyme inhibitors, and digoxin in all patients, were withheld 24 hours before the study. Patients with edema,

Results

Resting forearm blood flows did not differ between patients with CHF and normal subjects before infusion of $l$ -NMMA (2.1 ± 0.3 vs 2.8 ± 0.3 ml/min/100 ml) or after infusion of $l$ -NMMA (2.1 ± 0.3 vs 2.1 ± 0.3 ml/min/100 ml). $l$ -NMMA significantly decreased resting forearm blood flow in normal subjects (p <0.05) but not in patients with CHF.

To achieve a comparable degree of vasodilation in the 2 groups, mean doses of acetylcholine and nitroglycerin administered to patients with CHF were

Discussion

The present study demonstrates that during systemic inhibition of cyclooxygenase with indomethacin, regional inhibition of nitric oxide synthase with $l$ -NMMA significantly decreased the vasodilatory response to acetylcholine in normal subjects but not in patients with CHF. The presence of a noncyclooxygenase, non-nitric-oxide relaxing factor indicates that EDHF, rather than nitric oxide, may be the predominant endothelium-derived substance mediating vasodilation in response to acetylcholine in

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In addition, neurovascular coupling, as well as the basal tone of the retinal vasculature, which is determined by the summation of both metabolic and myogenic regulatory mechanisms, also play a role (Lecrux and Hamel, 2011; Pournaras et al., 2008; Flammer and Orgül, 1998). The DTM, on the other hand, uses reactive hyperaemia after a period of ischemia and, therefore, its output is strongly dependent on the balance between endothelial NO synthase (eNOS) and cyclooxygenase (COX) inhibition which attenuates the acetylcholine (Ach)-induced, endothelium- dependent, cutaneous vasodilatation (Katz and Krum, 2001). Other responsible factors also include a non-NO, non-prostanoid-dependent pathway, potentially attributable to the endothelium-derived hyperpolarizing factor (Ludmer et al., 1986) (EDHF).
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Thus, the NO signaling and/or EDHF system might represent a sex-specific target for preventive and therapeutic strategies. For example, altered EDHF responses may contribute to [9], or compensate for [10] ED in a sex-specific matter. It has been shown that EDHF contributes to sex-related differences in blood pressure control.
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This review aims to give an overview of the recent advances in understanding the importance of sex specific observations in endothelial maintenance, both in healthy and diseased conditions. The female endothelium is highlighted in the context of polycystic ovary syndrome and pre-eclampsia. Furthermore, sex differences are explored in chronic kidney disease, which is currently appreciated as one of public health priorities.
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In cardiovascular diseases, endothelial dysfunctions have been reported, where the release of NO, one of the important endothelium-derived vasodilators is compromised [15–17]. In many studies, the EDH-type response has been proposed to be enhanced in diseases, acting as a ‘back up’ system to compensate the loss of NO in the endothelium [15,18–20]. Studies in young, healthy human subjects, rat aorta, and rat cerebral arteries have shown that males exhibit a greater oxidative stress compared to females [5,21–23].
The present study examined whether vascular function, expression and activity of NADPH oxidases differ between sexes in porcine isolated coronary arteries (PCAs) using selective Nox inhibitors, ML-171 and VAS2870. Vascular responses of distal PCAs were examined under myographic conditions in the presence of a range of inhibitors. Nox activity in PCA homogenates was assessed using lucigenin-enhanced chemiluminescence. Protein expression of Nox1, Nox2 and Nox4 was compared using Western immunoblotting. The presence of ML-171 or DPI had no effect on the bradykinin-induced vasorelaxation in PCAs from females. In males, DPI shifted the EC₅₀ 2.8-fold to the right. In the presence of L-NAME and indomethacin, DPI and ML-171 had no effect in females, but enhanced the bradykinin-induced vasorelaxation in males. ML-171 had no effect on the forskolin-induced vasorelaxation but decreased the potency of U46619-induced tone in both sexes in the absence or presence of endothelium. VAS2870 had no effect on the bradykinin-induced vasorelaxation in both sexes but reduces the EDH-type response in males only. Nox activity was reduced by DPI and ML-171, but not VAS2870 in PCAs from both sexes. Protein expression of Nox1 and Nox2 in PCAs was higher in males compared to females whereas Nox4 was higher in females. Inhibition of Nox with ML-171 enhances while VAS2870 reduces the EDH-type response in PCAs from males but not females. This indicates that Nox-generated ROS play a role in the EDH-type response in males with differences attributed to the differential expression of Nox isoforms. This may underlie the greater oxidative stress observed in males.
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The fact that l-NAME blunted the response to ACh in the MI group suggests that either 1) NO function was not completely obliterated in the MI animals, or that 2) MI may blunt the ACh-induced release of other factors, such as endothelium-derived hyperpolarizing factor (EDHF). Indeed, Katz and Krum (2001) recently suggested that EDHF, rather than NO, played a larger role in the vasodilatory response to ACh in patients with heart failure. Because our focus was on the influence of endothelial dysfunction on functional vasodilation, we did not fully investigate the mechanisms of endothelial dysfunction in these animals.
Diminished bioavailability of nitric oxide (NO) may impair skeletal muscle arteriolar function after myocardial infarction (MI). We tested the hypotheses that chronic MI induced would diminish 1) endothelial function in large (resting diameter ~ 75 μm) feed arterioles, and 2) functional dilation in feed arterioles, but not smaller arcade (~ 25 μm) or transverse (~ 15 μm) arterioles, in the spinotrapezius muscle of female Sprague–Dawley rats. Additionally, we hypothesized that blockade of NO production with N^G-nitro-l-arginine methyl ester (l-NAME; 30 mg/kg i.v.) would have a greater blunting effect on control rats than MI rats. Endothelial function of the feed arterioles was assessed with an infusion of acetylcholine (1.5 μg i.v.) after pretreatment with indomethacin (5 mg/kg i.p.). MI blunted the response to acetylcholine in feed arterioles (p = 0.037), but did not affect resting or post-contraction diameter at any branching order. l-NAME had similar effects on MI and SHAM rats; the response to acetylcholine was blunted in feed arterioles (p = 0.003), resting diameter was diminished in arcade arterioles (p = 0.003), and post-contraction diameter was diminished in both arcade arterioles (p = 0.03) and transverse arterioles (p = 0.05). In conclusion, despite endothelial dysfunction in feed arterioles, functional dilation was not affected by MI in any branching order studied. l-NAME had similar effects on MI and SHAM rats that were branch order-dependent. These branch-order effects should be considered in future studies of the control of blood flow.
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These findings suggest that EETs sustain endothelium-dependent vasodilation in hypertension. A similar compensatory role for EETs may exist in patients with congestive heart failure (Katz and Krum, 2001). Forearm blood flow responses to ACH were decreased by COX and NOS inhibition in the normal subjects but not in patients with congestive heart failure.
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^☆: This study was supported by an investigatorship award from the American Heart Association, New York City Affiliate, New York, New York; by Grant NHLBI R29 HL51433 from the National Institutes of Health, Bethesda, Maryland; by the National Heart Foundation of Australia Clinical Research Program, and Grant 5 M01RR00645, Division of Research Resources, General Clinical Research Program, National Institutes of Health, Bethesda, Maryland.

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Acetylcholine-mediated vasodilation in the forearm circulation of patients with heart failure: indirect evidence for the role of endothelium-derived hyperpolarizing factor☆

Abstract

Section snippets

Study group

Results

Discussion

Biochem Pharmacol

J Am Coll Cardiol

Methods Enzymol

Life Sci

J Am Coll Cardiol

Am J Hypertens

Endothelium-derived hyperpolarizing factor and endothelium-dependent relaxations

Am J Respir Cell & Molec Biol

The importance of the hyperpolarizing mechanism increases as the vessel size decreases in endothelium-dependent relaxations in rat mesenteric circulation

J Cardiovasc Pharmacol

Heterogeneous distribution of endothelium-dependent relaxations resistant to NG-nitro-l-arginine in rats

Am J Physiol

Endothelium-dependent vasodilation is attenuated in patients with heart failure

Circulation

Heterogeneous distribution of endothelium-dependent relaxations resistant to NG-nitro- $l$ -arginine in rats