Mesenteric Vasoconstrictor Responses Research Articles

Effects of chronic nitric oxide synthase inhibition on the cardiovascular responses to cannabinoids in vivo and in vitro

Since the vasorelaxant potency of the endocannabinoid anandamide is enhanced in perfused mesenteric vascular beds from rats made hypertensive by chronic inhibition of NO synthase (L-NAME in drinking water), we hypothesized that in vivo, anandamide-induced vasodilatation would be similarly enhanced in L-NAME-treated animals. Male Sprague-Dawley rats were given L-NAME in drinking water (7.5 mg kg(-1) day(-1)) for 4 weeks. Relaxant effects of anandamide were measured in perfused mesenteric vascular beds and in isolated small mesenteric arteries. Renal, mesenteric and hindquarters haemodynamic responses to anandamide, methanandamide, the synthetic cannabinoid agonist WIN-55212-2 and the cannabinoid receptor antagonist AM251 were assessed in conscious, chronically-instrumented rats. Vasorelaxant responses to anandamide were enhanced in the perfused mesentery but not in isolated mesenteric resistance vessels. In vivo, anandamide caused vasodilatation only in the hindquarters vascular bed and only in control rats. Methanandamide caused a late-onset (40 min after administration) tachycardia, mesenteric and hindquarters vasoconstriction, and renal vasodilatation, which did not differ between control and L-NAME-treated rats. AM251 had no effect on resting blood pressure in control or L-NAME-treated rats and WIN55212-2 caused pressor and renal and mesenteric vasoconstrictor responses, with hindquarters vasodilatation in both groups of animals. The results provide no in vivo evidence for enhanced vasodilator responses to cannabinoids, or up-regulation of endocannabinoids or their receptor activity, following chronic NO synthase inhibition.

Induction of insulin resistance by high‐sucrose feeding does not raise mean arterial blood pressure but impairs haemodynamic responses to insulin in rats

1. This study was undertaken to further investigate the effects of a sucrose-enriched diet on vascular function and insulin sensitivity in rats. 2. Male Sprague-Dawley rats were randomized to receive a sucrose- or regular rat chow-diet for 4 weeks. A first group of sucrose- and chow-fed rats was instrumented with pulsed Doppler flow probes and intravascular catheters to determine blood pressure, heart rate, regional blood flows and insulin sensitivity in conscious rats. Insulin sensitivity was assessed by the euglycemic hyperinsulinemic clamp technique. Glucose transport activity was examined in isolated muscles by using the glucose analogue [(3)H]-2-deoxy-D-glucose. A second group of sucrose- and chow-fed rats was used to obtain information regarding nitric oxide synthase (NOS) isozymes protein expression in muscles, and determine endothelin content in vascular tissues isolated from both dietary groups. 3. Sucrose feeding was found to induce insulin resistance, but had no effect on resting blood pressure, heart rate, or regional haemodynamics. This insulin resistance was accompanied by alteration in the vascular responses to insulin. Insulin-mediated skeletal muscle vasodilation was impaired, whereas the mesenteric vasoconstrictor response was potentiated in sucrose-fed rats. A reduction in eNOS protein content in muscle and an increase in vascular endothelin peptide were noted in these animals. Moreover, a reduction in insulin-simulated glucose transport activity was also noted in muscles isolated from sucrose-fed rats. 4. Together these data suggest that a cluster of metabolic and haemodynamic abnormalities occur in response to the intake of simple sugars in rats.

Neuropeptide Y Y1 receptor mediated mesenteric vasoconstriction in the pig in vivo

The object of the present study was to investigate the effects of the sympathetic cotransmitter neuropeptide Y (NPY), and the closely related gut hormone peptide YY (PYY), on splanchnic blood flow regulation in the anaesthetized pig in vivo. Systemic injections of NPY, PYY and the NPY Y 1 receptor agonist [Leu 31Pro 34]NPY (470 pmol kg −1 each) evoked pressor and mesenteric vasoconstrictor responses that were largely abolished by the selective NPY Y 1 receptor antagonist H 409/22 (60 nmol kg −1 min −1). In contrast, the NPY Y 2 receptor agonist N-acetyl[Leu 28Leu 31]NPY(24-36) (1.1 nmol kg −1), a dose of which potently evoked splenic NPY Y 2 receptor mediated (not affected by H 409/22) vasoconstriction, did not evoke any mesenteric vascular response. Mesenteric vascular responses to angiotensin II (10 pmol kg −1), α,β-methylene ATP (10 nmol kg −1) and the α 1-adrenoceptor agonist phenylephrine (15 nmol kg −1), were not inhibited by H 409/22. It is concluded that NPY and PYY evokes porcine mesenteric vasoconstriction mediated by the NPY Y 1 receptor subtype, as demonstrated by selective and specific inhibition exerted by the NPY Y 1 receptor antagonist H 409/22, in vivo.

Mesenteric vasoconstrictor response to 5-hydroxytryptamine in the in situ blood autoperfused rat mesentery: involvement of 5-HT 2B and/or 5-HT 2C receptor activation

Using a number of agonist and antagonist compounds, we attempted to characterize the responses and receptors involved in the effects of 5-hydroxytryptamine (5-HT) in the in situ blood perfused rat mesentery. An intra-arterial (i.a.) bolus injection of 5-HT increased mesenteric perfusion pressure in a dose-dependent way but did not change the systemic blood pressure. The selective 5-HT 2 receptor agonists α-methyl-5-HT, 1-(3-chlorophenyl)piperazine dihydrochloride ( m-CPP) and (±)-1-(4-iodo-2,5-dimethoxyphenyl)-2-aminopropane hydrochloride (DOI), caused a local vasoconstrictor effect in the autoperfused vascular mesenteric bed. Intra-arterial injection of 5-carboxamidotryptamine (5-CT) and 1-( m-chlorophenyl)-biguanide ( m-CPBG) did not modify the mesenteric perfusion pressure. The vasoconstrictor effect elicited by 5-HT and α-methyl-5-HT was significantly decreased by ritanserin and by a selective 5-HT 2B/2C receptor antagonist, N-3-pyridinyl-3,5-dihydro-5-methyl-benzo[1,2- b:4,5- b′]dipyrrole-1(2 H)-carboxamide hydrochloride (SB 206553), but was not modified by prazosin, propranolol, indomethacin or enalapril pretreatment. Our data suggest that the vasoconstrictor serotonergic response induced in the in situ autoperfused rat mesenteric vascular bed is mainly mediated by activation of 5-HT 2B and/or 5-HT 2C receptors.

Cytochrome P450 metabolites of arachidonic acid may be important mediators in angiotensin II-induced vasoconstriction in the rat mesentery in vivo

We have investigated the role of cytochrome P450 (CYP-450) metabolites of arachidonic acid in the modulation of vascular reactivity to angiotensin II in vivo using an in situ blood-perfused mesenteric preparation in anaesthetized spontaneously hypertensive rats (SHR). Miconazole, a non-selective inhibitor of CYP-450 that inhibits both hydroxylation and epoxidation, substantially suppressed mesenteric vasoconstrictor responses to angiotensin II in SHR, but had no effect on responses to noradrenaline or sympathetic nerve stimulation. In normotensive Wistar–Kyoto (WKY) rats, miconazole caused only a modest suppression of vasoconstrictor responses to angiotensin II. N-Methylsulphonyl-12,12-dibromododec-11-enamide (DDMS), a new selective inhibitor of CYP-450 ω-hydroxylase activity, decreased mean intra-arterial blood pressure and significantly attenuated mesenteric angiotensin II-induced vasoconstrictor responses in SHR. Isolated mesenteric vessels were able to metabolize 14C-labelled arachidonic acid to hydroxyeicosatetraenoic acids (HETEs) in vitro, and this was substantially inhibited by DDMS. The results from the present studies combined with the existing evidence that angiotensin II stimulates the release of 20-HETE, a CYP-450 metabolite of arachidonic acid, suggest that CYP-450-derived HETEs may be important mediators in angiotensin II-induced vasoconstriction. However, the development of more sensitive assays for the detection in vivo of 20-HETE in mesenteric vessels would be required to confirm these findings.

Cytochrome P450 metabolites of arachidonic acid may be important mediators in angiotensin II-induced vasoconstriction in the rat mesentery in vivo

We have investigated the role of cytochrome P450 (CYP-450) metabolites of arachidonic acid in the modulation of vascular reactivity to angiotensin II in vivo using an in situ blood-perfused mesenteric preparation in anaesthetized spontaneously hypertensive rats (SHR). Miconazole, a non-selective inhibitor of CYP-450 that inhibits both hydroxylation and epoxidation, substantially suppressed mesenteric vasoconstrictor responses to angiotensin II in SHR, but had no effect on responses to noradrenaline or sympathetic nerve stimulation. In normotensive Wistar-Kyoto (WKY) rats, miconazole caused only a modest suppression of vasoconstrictor responses to angiotensin II. N-Methylsulphonyl-12, 12-dibromododec-11-enamide (DDMS), a new selective inhibitor of CYP-450 omega-hydroxylase activity, decreased mean intra-arterial blood pressure and significantly attenuated mesenteric angiotensin II-induced vasoconstrictor responses in SHR. Isolated mesenteric vessels were able to metabolize (14)C-labelled arachidonic acid to hydroxyeicosatetraenoic acids (HETEs) in vitro, and this was substantially inhibited by DDMS. The results from the present studies combined with the existing evidence that angiotensin II stimulates the release of 20-HETE, a CYP-450 metabolite of arachidonic acid, suggest that CYP-450-derived HETEs may be important mediators in angiotensin II-induced vasoconstriction. However, the development of more sensitive assays for the detection in vivo of 20-HETE in mesenteric vessels would be required to confirm these findings.

Peroxynitrite-mediated attenuation of α- and β-adrenoceptor agonist-induced vascular responses in vivo

Peroxynitrite is produced by vascular endothelial and smooth muscle cells in response to inflammation, induces vascular relaxation, and alters vascular responses to endothelial-derived relaxing factors. The present study examined the changes in mean arterial pressure and hindquarter, renal, and mesenteric vascular resistances produced by the systemic administration of (i) the catecholamines epinephrine or norepinephrine, (ii) the α 1-adrenoceptor agonist phenylephrine, (iii) the β-adrenoceptor agonist isoproterenol or (iv) [Argδ] vasopressin in pentobarbital-anesthetized rats prior to and following the systemic administration of peroxynitrite. The systemic administration of peroxynitrite significantly inhibited (i) epinephrine-induced pressor and renal and mesenteric vasoconstrictor responses, (ii) norepinephrine-induced pressor and hindquarter, renal, and mesenteric vasoconstrictor responses, (iii) phenylephrine-induced hindquarter and mesenteric vasoconstrictor responses, and (iv) isoproterenol-induced depressor and hindquarter and renal vasodilator responses. In comparison, the systemic administration of peroxynitrite had no effect on arginine vasopressin-induced pressor or vasoconstrictor responses. These results demonstrate selective and consequential attenuation of the hemodynamic effects produced by α- and β-adrenoceptor agonists, suggesting that selective impairment of adrenoceptors by peroxynitrite may play a critical role in the hemodynamic dysfunction associated with inflammatory conditions.

Neuropeptide Y and mesenteric sympathetic vasoconstriction in pregnant and non-pregnant Wistar-Kyoto rats.

1. We have investigated the role of neuropeptide Y (NPY) in sympathetic vasoconstriction and its possible contribution to decreased mesenteric vasoconstrictor responses to electrical field stimulation (EFS) in pregnancy in vivo using an in situ bloodperfused mesenteric preparation in 18-20 day pregnant and age-matched non-pregnant Wistar-Kyoto (WKY) rats. 2. Mean blood pressure (BP) was decreased in pregnant compared with non-pregnant rats. Mesenteric basal perfusion pressure (PP) was not significantly reduced in pregnancy. BIBP 3226, a specific NPY Y1 receptor antagonist, did not affect mean BP or mesenteric basal PP in pregnant or non-pregnant animals. 3. Mesenteric vasoconstrictor responses to EFS were blunted in pregnant compared with non-pregnant controls. BIBP 3226 significantly suppressed mesenteric vasoconstrictor responses to EFS in both pregnant and non-pregnant animals. Decreased mesenteric responses to EFS in pregnancy were still evident following NPY Y1 receptor blockade. 4. These results suggest that endogenous NPY plays a role in rat mesenteric sympathetic vasoconstriction in vivo. However, NPY is unlikely to be responsible for the impairment of EFS-induced mesenteric vasoconstriction in pregnant WKY rats.

Differential effects of endotoxaemia on pressor and vasoconstrictor actions of angiotensin II and arginine vasopressin in conscious rats.

1. Regional haemodynamic responses to arginine vasopressin (AVP; 0.5, 1.0, 5.0 pmol i.v.) and angiotensin II (AII; 5.0, 10.0, 50.0 pmol i.v.) were measured in conscious Long Evans rats at various times (0, 2, 6 and 24 h) during infusion of lipopolysaccharide (LPS, 150 microg kg(-1) h(-1), i.v., n=9) or saline (n=9). Additional experiments were performed in vasopressin-deficient (Brattleboro) rats infused with LPS (n=7) or saline (n=8) to determine whether or not, in the absence of circulating vasopressin, responses to the exogenous peptides differed from those in Long Evans rats. 2. In the Long Evans rats, during the 24 h infusion of LPS, there was a changing haemodynamic profile with renal vasodilatation from 2 h onwards, additional mesenteric vasodilatation at 6 h, and a modest hypotension (reduction in mean arterial blood pressure (MAP) from 103+/-1 to 98+/-2 mmHg) associated with renal and hindquarters vasodilatation at 24 h. 3. In the Brattleboro rats, the changes in regional haemodynamics during LPS infusion were more profound than in the Long Evans rats. At 2 h and 6 h, there was a marked fall in MAP (from 103+/-3 mmHg; to 65+/-3 mmHg at 2 h, and to 82+/-4 mmHg at 6 h) associated with vasodilatation in all three vascular beds. After 24 h infusion of LPS, the hypotension was less although still significant (from 103+/-3 mmHg; to 93+/-4 mmHg, a change of 10+/-4 mmHg), and there was renal and hindquarters vasodilatation, but mesenteric vasoconstriction. 4. During infusion of LPS, at each time point studied, and in both strains of rat, pressor responses to AII and AVP were reduced, but the changes were less marked at 6 h than at 2 h or 24 h. The reduced pressor responses were not accompanied by generalized reductions in the regional vasoconstrictor responses. Thus, in the Long Evans rats, the renal vasoconstrictor responses to both peptides were enhanced (at 6 h and 24 h for AVP; at all times for AII), whereas the mesenteric vasoconstrictor response to AVP was unchanged at 2 h, enhanced at 6 h and reduced at 24 h. The mesenteric vasoconstrictor response to AII was reduced at 2 h, normal at 6 h and reduced at 24 h. The small hindquarters vasoconstrictor responses to both peptides were reduced at 2 h and 6 h, but normal at 24 h. 5. In the Brattleboro rats, the renal vasoconstrictor responses to both peptides were reduced at 2 h and enhanced at 6 h and 24 h, whereas the mesenteric vasoconstrictor response to AVP was normal at 2 h and 6 h, and reduced at 24 h. The response to AII was reduced at 2 h, normal at 6 h and reduced again at 24 h. There were no reproducible hindquarters vasoconstrictions to AVP in the Brattleboro rats. The small hindquarters vasoconstrictor responses to AII were unchanged at 2 h and enhanced at 6 h and 24 h. 6. In isolated perfused mesenteric vascular beds, removed after 24 h of LPS infusion in vivo, there was an increase in the potency of AVP in both strains (Long Evans, ED50 saline: 56.9+/-15.0 pmol, ED50 LPS: 20.4+/-4.8 pmol, Brattleboro, ED50 saline: 38.6+/-4.2, ED50 LPS: 19.6+/-2.9 pmol), but no change in the responses to AII. 7. These findings indicate that a reduced pressor response to a vasoconstrictor challenge during LPS infusion is not necessarily associated with a reduced regional vasoconstriction. The data obtained in the Brattleboro rats indicate a potentially important role for vasopressin in maintaining haemodynamic status during LPS infusion in Long Evans rats. However, it is unlikely that the responses to exogenous AVP (or AII) are influenced by changes in the background level of endogenous vasopressin, since the patterns of change were similar in Long Evans and Brattleboro rats. 8. The results obtained in isolated perfused mesenteric vascular beds differed from those in vivo, possibly due to the conditions pertaining with in vitro perfusion.

Effects of the antihypertensive agent, cicletanine, on noradrenaline release and vasoconstriction in perfused mesenteric artery of SHR.

1. The mechanism by which cicletanine (CIC) exerts its antihypertensive effects has not been fully elucidated. The present study was undertaken to examine the effects of in vivo and in vitro treatment with CIC on the pressor response and noradrenaline (NA) overflow during periarterial nerve stimulation (PNS) in perfused mesenteric arterial beds isolated from spontaneously hypertensive rats (SHR). 2. CIC at a dose of 50 mg kg(-1) day(-1) was administered orally to both SHR and normotensive Wistar-Kyoto rats (WKY) from the 6th to 10th week of age. At the 10th week, the isolated mesenteric arterial bed was perfused with Krebs-Henseleit buffer and changes in perfusion pressure and NA overflow during PNS were measured. 3. Chronic treatment with CIC suppressed the age-related elevation of systemic blood pressure in SHR but not in WKY. 4. The PNS (20 Hz)-induced mesenteric vasoconstrictor response and NA overflow were greater in SHR than in WKY. In the vasculature of SHR chronic treatment with CIC resulted in a significant attenuation of the vasoconstriction and the NA overflow during PNS, whereas it did not alter vasoconstrictor responses to bolus injections of KCl and phenylephrine. 5. Treatment with 30 microM CIC in vitro diminished the PNS-induced vasoconstriction and NA overflow but not the NA- and KCl-induced vasoconstriction in the vasculature of untreated SHR. 6. In the vasculature of SHR PNS-induced NA overflow was attenuated by prostaglandin E2 (0.05 microM), whereas it was augmented by the cyclo-oxygenase inhibitor diclofenac-Na (30 microM). In the presence of diclofenac, in vitro treatment with CIC did not attenuate the NA overflow during PNS. 7. The results suggest that the antihypertensive effect of CIC in SHR is partially due to the presynaptic inhibition of NA release during sympathetic nerve activation. Transjunctional inhibition of NA release by prostaglandins may contribute to the inhibitory action of CIC on NA release in the vasculature of SHR.

Synergistic vascular effects of dietary sodium supplementation and angiotensin II administration.

The hypertensinogenic action of dietary sodium supplementation was investigated in angiotensin II (ANG II)-treated rats. We hypothesized that high-sodium diet would potentiate ANG II-induced vasoconstriction and hypertension, including the development of structural vascular changes, through synergistic action with ANG II in stimulating sympathetic activity and vascular growth. Mesenteric vasoconstrictor responses to ANG II, norepinephrine (NE), arginine vasopressin (AVP), and periarterial nerve stimulation were measured in Sprague-Dawley rats after 7-10 days of the following treatments: 200 ng.kg-1.min-1 ip ANG II (n = 12), 4% NaCl diet (n = 11), ANG II + 4% NaCl diet (n = 7), and 0.7% NaCl diet (controls) (n = 15). Additional rats received 50 ng.kg-1.min-1 sc ANG II (n = 8), 2% NaCl diet (n = 9), ANG II + 2% NaCl diet (n = 6), or 0.7% NaCl diet (controls) (n = 10) for 12 wk. Systolic blood pressure (SBP) values were measured weekly for 4 wk and then every other week for 8 wk. Then, the wall-to-lumen ratio (W/L) of mesenteric resistance arteries (< 150 microns OD) was measured after in situ perfusion-fixation. After 7-10 days of treatment, there were no significant changes in SBP in any of the groups. High-sodium diet increased vasoconstrictor responses to ANG II (P < 0.01) and nerve stimulation (P < 0.02), but not to NE or AVP, and in combination with ANG II treatment further potentiated vasoconstrictor responses to ANG II (synergism). After 12 wk of treatment, ANG II increased W/L of small resistance arteries by 11% (P < 0.05) without a significant rise in SBP. ANG II and 2% NaCl diet in combination raised SBP by 36 mmHg (P < 0.01) and increased small artery W/L by 28% (P < 0.001) compared with values obtained in control rats. To test the specificity of the interaction between ANG II and high-sodium diet, all the experiments were repeated during phenylephrine (PE, 10 micrograms.kg-1.min-1 sc) treatment of rats. PE by itself or in combination with high-sodium diet had no effect on the measured parameters. Thus short-term administration of high-sodium diet appears to potentiate vasoconstrictor responses to ANG II by facilitating sympathetic neurotransmission, and long-term administration of high-sodium diet raises SBP by potentiating the trophic vascular effects of ANG II. The interaction appears to be specific to ANG II and is occurring on the vascular level.

Effects of 17 beta-oestradiol on vascular responses in the in situ blood-perfused mesentery of Wistar-Kyoto rats.

1. To determine whether endogenous oestrogen plays a role in pregnancy induced decreased vascular reactivity we have examined the effects of 17 beta-oestradiol on vasoconstrictor responses to various stimuli using an in situ blood-perfused mesenteric vascular preparation in Wistar-Kyoto (WKY) rats. 2. Daily administration of 17 beta-oestradiol (500 micrograms/kg, s.c.) for 15 days significantly enhanced mesenteric vasoconstrictor responses to noradrenaline (NA), without affecting responses to the electrical stimulation of sympathetic nerves (ES) and angiotensin II (AngII). 3. Nitric oxide (NO) synthesis inhibition by nitro-L-arginine methyl ester (L-NAME; 100 mg/kg, i.v.) significantly potentiated mesenteric vasoconstrictor responses to all stimuli in both 17 beta-oestradiol-treated and control animals. The difference in NA responses between groups was diminished following NO synthesis inhibition. 4. These findings do not support the hypothesis that increased endogenous oestrogen plays a role in decreased mesenteric vascular reactivity in pregnancy. However, responses to oestrogen may be dose-dependent and enhancement of vasoconstrictor responses to NA may be relevant to oral contraceptive-induced hypertension.

Characterization of α-adrenoceptor activity in the preterm piglet mesentery

To characterize neonatal mesenteric α-adrenoceptor populations, an extracorporeal perfusion circuit was established to control intestinal blood flow in prematurely delivered (by cesarean section at 90% of gestational age) piglets. Activation of α 1-adrenoceptors was documented by observing dose-dependent increases in mesenteric perfusion pressure after intramesenteric arterial injection of methoxamine; α 2-adrenoceptor activity was confirmed by finding similar increases in mesenteric perfusion pressure after intramesenteric arterial injections of BHT 933. Peripheral intravenous injections of WB 4101 (a competitive α 1A-adrenoceptor antagonist), but not clorethylclonidine (CEC, an α 1B-adrenoceptor antagonist), significantly blunted ( P < .05, ANOVA) the mesenteric vasoconstrictor responses to methoxamine. The mesenteric vasoconstrictor response to BHT 933 (an α 2-adrenoceptor agonist) also was blunted by WB 4101, but not by CEC. These data suggest that α 1A- and α 2-adrenoceptors can be activated in the small intestinal mesentery of piglets well before they reach full-term maturation, although receptor specificity has not been fully established.

Characterization of alpha-adrenoceptor activity in term-newborn piglet mesentery.

For further characterization of neonatal mesenteric alpha 1-adrenoceptor populations, an extracorporeal perfusion circuit was established to control intestinal blood flow in 0-2 day old piglets. Activation of alpha 1-adrenoceptors was first documented by observing dose-dependent increases in mesenteric perfusion pressure after intra-mesenteric arterial injection of methoxamine and noradrenaline. Peripheral intravenous injections of WB 4101 (a competitive alpha 1A-adrenoceptor antagonist), but not clorethylclonidine (CEC, an alpha 1B-adrenoceptor antagonist), significantly (P < 0.05, analysis of variance) blunted mesenteric vasoconstrictor responses to those agonists. That the mesenteric vasoconstrictor response to mesenteric plexus stimulation was unaltered by CEC, but was muted by both WB 4101 and SK&F 104856 (a post-junctional alpha 1- and alpha 2-adrenoceptor antagonist) suggests that pre- and post-junctional alpha 1A-adrenoceptors are present and functional at birth.

The peroxynitrite product 3-nitro- l-tyrosine attenuates the hemodynamic responses to angiotensin II in vivo

Peroxynitrite is a potent oxidant formed endogenously by the near diffusion-limited reaction of nitric oxide with superoxide anion. Peroxynitrite specifically adds a nitro group to the ortho position of the phenolic ring of free and protein-associated tyrosines to form the stable product 3-nitro- l-tyrosine. Systemic administration of 3-nitro- l-tyrosine markedly inhibits the subsequent hemodynamic responses to α 1- and β-adrenoceptor agonists in anesthetized rats. Angiotensin II is an important modulator of vascular tone. The vasoconstrictor effects of this hormone are known to involve the release of catecholamines from sympathetic tissues. In the present study, we examined whether 3-nitro- l-tyrosine (2.5 μmol/kg i.v.) would attenuate the hemodynamic responses produced by angiotensin II (0.1–1.0 μg/kg i.v.). Angiotensin II produced increases in mean arterial pressure, and renal and mesenteric vascular resistances, but no changes in hindquarter vascular resistance. The pressor and renal and mesenteric vasoconstrictor responses produced by angiotensin II were significantly attenuated 30–60 min following the administration of 3-nitro- l-tyrosine. Further attenuation of these responses was evident 120–180 min following the administration of 3-nitro- l-tyrosine. The α 1-adrenoceptor antagonist prazosin also diminished the pressor and renal and mesenteric vasoconstrictor responses produced by angiotensin II. These results demonstrate that 3-nitro- l-tyrosine inhibits the hemodynamic responses to angiotensin II, possibly through the inhibition of α 1-adrenoceptor-mediated events. The effect of 3-nitro- l-tyrosine on the hemodynamic action of angiotensin II raises the possibility that 3-nitro- l-tyrosine may be involved in the pathogenesis of the hemodynamic disturbances associated with inflammatory conditions, such as atherosclerosis, ischemia-reperfusion, and sepsis, where formation of peroxynitrite is favored.

Endotoxin-induced inhibition of mesenteric vasodilator responses to acetylcholine, bradykinin, and post-occlusion hyperemia in anesthetized rats.

The effects of endotoxin (20 mg kg-1 i.p.) on the mesenteric vascular responses to acetylcholine, bradykinin, sodium nitroprusside, and to transient occlusion of the superior mesenteric artery were examined in rats anesthetized with pentobarbitone. Mesenteric vasodilator responses to close arterial injections of acetylcholine and bradykinin were reduced at 1.5 h after endotoxin and almost abolished by 4 h; responses to sodium nitroprusside were unaffected. Occlusion of the superior mesenteric artery for 30, 60, or 120 s produced, on release of the occlusion, a time-dependent vasodilator response in the mesenteric circulation (post-occlusion hyperemia). This hyperemia was markedly reduced by nitro-L-arginine methyl ester (L-NAME); L-NAME did not modify acetylcholine-induced vasodilation. Endotoxin-pretreatment did not modify mesenteric post-occlusion hyperemia 1.5 h after administration but markedly reduced the response by 2.5 h. The administration of L-NAME to endotoxin-treated rats did not further attenuate the hyperemic responses. Mesenteric vasoconstrictor responses to phenylephrine were not modified by endotoxin, although systemic pressor responses to this agent were impaired. We concluded that endotoxin impairs endothelium and nitric oxide-dependent vasodilator responses in the mesenteric circulation.

Baroreceptor modulation of regional haemodynamic responses to acute stress in rat

We examined the effects of chronic sinoaortic denervation (SAD) on regional haemodynamic responses to acute environmental stress in rats. In conscious male intact ( n = 12) and SAD (2 weeks before study, n = 7) rats, arterial pressure and blood flow velocities (pulsed Doppler probes) in the subdiaphragmatic aorta, superior mesenteric artery and distal aorta (hindquarters) were simultaneously recorded. In response to air jet stress, intact rats showed modest increases in arterial pressure that were accompanied by vasoconstriction in the mesentery and vasodilatation in the hindquarters. These regional haemodynamic changes were almost balanced, as indicated by the lack of change in the subdiaphragmatic aortic conductance. SAD markedly enhanced the pressor and mesenteric vasoconstrictor responses and blunted the hindquarters vasodilatation. After acute β-adrenoceptor blockade with propranolol, the stress-induced hindquarters vasodilatation was strongly reduced in the intact rats and was reversed into vasoconstriction in the SAD rats. These results point to an opposing interaction between centrally-induced sympathoexcitation and arterial baroreceptor reflex activation during stress. This probably favours the hyperaemic response in the skeletal muscles at a lower metabolic cost.

Effect of Maturation on α-Adrenoceptor Activity in Newborn Piglet Mesentery

To characterize the mesenteric α1- and α2-adrenoceptor populations in newborn piglets, an extracorporeal circuit was established to control intestinal blood flow in 0- to 2-day old and 10- to 14-day old animals. In both groups, α-adrenoceptor activation was first documented by observing dose-dependent increases in mesenteric perfusion pressure after intramesenteric arterial injection of α-adrenoceptor agonists. In the 10-to 14-day old piglets, mesenteric vasoconstrictor responses to α1-adrenoceptor agonists (methoxamine and norepinephrine) and an α2-adrenoceptor agonist (BHT-933) were each blunted (P< 0.05, analysis of variance) by peripheral intravenous injections of prazosin (an α1-adrenoceptor antagonist) and yohimbine (an α2-adrenoceptor antagonist), respectively. The mesenteric vasoconstrictor responses to those agonists were not significantly attenuated by prazosin or yohimbine in 0- to 2-day old animals, nor were they blunted by YM-12617 (α1-adrenoceptor antagonist) or idazoxan (α2-adrenoceptor antagonist)—compounds that are structurally unrelated to prazosin and yohimbine, respectively. In addition, mesenteric vasoconstrictor responses to other known vasoconstrictor agents—angiotensin II, neuropeptide Y, and a thromboxane A2mimic (U-46619)—were not effected in either age group by prazosin or yohimbine, implying these agents act independently of α-adrenoceptor mechanisms. These data suggest that (1) there exists functional mesenteric α1- and α2-adrenoceptor-like activity in 10- to 14-day old piglets that, in 0- to 2-day old animals, is not specifically expressed; and (2) mesenteric α-adrenoceptor function becomes more selective as newborn piglets mature.

INCREASING RESTING VASCULAR TONE IN CONSCIOUS DOGS DOES NOT ALTER THE MESENTERIC VASOCONSTRICTOR RESPONSES TO ANP

1. There is considerable in vitro evidence that, at high concentrations, atrial natriuretic peptide (ANP) acts directly on pre-constricted blood vessels to cause vasorelaxation. Previously, we have seen vasoconstriction rather than vasodilatation in conscious dogs at physiological levels of the peptide. It is possible that the low resting vascular tone in our conscious, unstressed animals prevented the manifestation of the relaxant properties of ANP in vivo. 2. In the present study in conscious, instrumented dogs, we studied the mesenteric vascular responses to 10 min infusions of ANP (10, 25, 50 and 100 ng/kg per min, i.v.) when resting vascular tone was enhanced with a continuous infusion of AVP (75 pg/kg per min, i.v.) and compared these with responses in the normal condition (no added AVP). 3. Mesenteric vascular resistance was increased by ANP (10, 25, 50 and 100 ng/kg per min) by 9 +/- 2, 20 +/- 6, 29 +/- 7 and 32 +/- 9%, respectively. Increased resting vascular tone did not alter the mesenteric vasoconstrictor response to ANP. Thus, the discrepancy between in vitro (vasorelaxation) and in vivo (vasoconstriction) findings may be the result of the widely different concentrations of ANP used, rather than the state of resting vascular tone.

Enhancement of the haemodynamic effects of NG‐monomethyl‐l‐arginine by transforming growth factor‐β1 in conscious, normal, but not endotoxaemic, rats

1. Male, Long Evans rats (350-450 g) were chronically instrumented for the measurement of regional haemodynamics, and the effects of TGF-beta 1 (25 micrograms kg-1 i.v. bolus) were assessed during infusion of saline (n = 9) or lipopolysaccharide (LPS, 150 micrograms kg-1 h-1; n = 12). In the same animals, responses to NG-monomethyl-L-arginine (L-NMMA 10 mg kg-1 bolus; 10 mg kg-1 h-1 infusion) were determined 18 h after administration of TGF-beta 1. In a separate experiment, the effects of the endothelin antagonist, SB 209670 (10 micrograms kg-1 min-1) on responses to TGF-beta 1 and to L-NMMA subsequently, were determined. 2. In the absence of LPS, TGF-beta 1 had slow-onset bradycardic and pressor effects accompanied by mesenteric and hindquarters, but not renal, vasoconstriction. Eighteen hours after TGF-beta 1, these effects had gone, but the bradycardic, pressor, and mesenteric vasoconstrictor responses to L-NMMA were enhanced. The haemodynamic changes following TGF-beta 1, and the augmentation of the subsequent responses to L-NMMA, were inhibited by SB 209670. These results are consistent with TGF-beta 1 stimulating the synthesis and release of endothelin, and an involvement of the latter in responses to L-NMMA. 3. The pressor effects of TGF-beta 1 were similar in LPS-infused and saline-infused animals, but in the former group the mesenteric vasoconstriction was enhanced and the hindquarters vasoconstriction diminished. Since, in the absence of TGF-beta 1, LPS-infused animals showed a developing hindquarters vasodilatation and mesenteric vasoconstriction, it is feasible that, in the presence of TGF-beta 1 and LPS together, the haemodynamic profile represented an amalgam of the individual effects of the two interventions, rather than a specific effect of TGF-beta 1 on the haemodynamic sequelae of endotoxaemia. 4. In the presence of LPS, haemodynamic responses to L-NMMA were suppressed, and TGF-beta 1 generally did not affect this suppression. A possible explanation of this observation is that LPS increased circulating endothelin levels, and thus resulted in desensitization to the effects of endothelin released following administration of L-NMMA.