Adrenergic, purinergic, and endothelial mediators and modulators of norepinephrine-induced mesenteric autoregulatory escape.

We evaluated the effects of potential factors in autoregulatory escape from norepinephrine-induced vasoconstriction in rat anterior mesenteric artery. We determined mesenteric artery blood flow velocity with a pulsed Doppler, sonic flowmeter, and systemic arterial blood pressure with a transducer. A 4-min norepinephrine infusion (0.125-1.0 x 10(-8) M/min) intravenously evoked a dose-dependent, initial vasoconstriction that was followed by rapid escape of blood flow toward or above the control value during sustained norepinephrine administration. Neonatal capsaicin treatment enhanced vasoconstrictor responses to norepinephrine but failed to affect escape parameters. Propranolol decreased norepinephrine-induced escape dose dependently. Adenosine deaminase attenuated escape, and the combination of this enzyme plus propranolol nearly abolished escape from norepinephrine-induced vasoconstriction. Methylene blue also diminished autoregulatory escape. These findings suggest that norepinephrine-induced autoregulatory escape involves simultaneous beta-adrenoceptor, purinergic, and endothelial mediation. Norepinephrine-evoked mesenteric vasoconstriction appears to involve predominantly alpha 2-adrenoceptors and is modulated by peptidergic sensory nerves and adenosine.

Multifactorial mediation of post norepinephrine induced intestinal hyperemia.

We evaluated several potential endogenous mediators of post norepinephrine induced hyperemia in the mesenteric circulation. Hyperemia was elicited in the anesthetized rat anterior mesenteric artery after cessation of intravenous norepinephrine infusion at 0.125 to 1.0 x 10(-8) M/min x 4 min. Arterial blood pressure was measured with a transducer, and the velocity of arterial blood flow was determined with pulsed Doppler velocimetry. Conductance at the height of mesenteric hyperemia, the post norepinephrine hyperemia volume, and the autoregulatory escape volume were calculated from recorded measurements. The higher doses of norepinephrine increased both the hyperemia volume and peak conductance in both control and capsaicin pretreated rats. Hyperemic parameters were significantly diminished by pretreatment with either yohimbine, propranolol, adenosine deaminase, or methylene blue. Combining adenosine deaminase with propranolol further reduced peak conductance and the hyperemia volume compared with enzyme pretreatment alone. The magnitude of hyperemia was related to the escape volume but not to the extent of norepinephrine induced vasoconstriction. We conclude that post norepinephrine induced hyperemia in the rat mesenteric circulation is modulated by alpha 2 and beta 2 adrenergic receptor activation, adenosine release, and endothelial factors.

Adenosine modulates reactive hyperemia in rat gut.

Intestinal reactive hyperemia is an abrupt blood flow increase following release from anterior mesenteric arterial occlusion. We investigated the role of adenosine in reactive hyperemia. In anesthetized rats, mesenteric arterial velocity of blood flow was determined with pulsed Doppler velocimetry and arterial pressure with a transducer. Three indices quantifying reactive hyperemias obtained following 30, 60, and 120 s arterial occlusions included duration, the volume of blood flow exceeding preocclusion blood flow, and the percentage increase in conductance. In six rat groups (half fasted and half with intrajejunal bile-oleate solutions), hyperemia parameters were determined before and after administration of either adenosine deaminase (ADA) or two adenosine receptor antagonists, namely 8-phenyltheophylline (8-PT) and 1,3-dipropyl-7-methylxanthine (DPMX). In fasted gut the three agents had variable effectiveness against reactive hyperemia, although 8-PT was the most consistent inhibitor. Instillation of intrajejunal lipid evoked a stable hyperemia and increased duration and blood flow volume after each occlusive period. ADA and 8-PT were more effective against reactive hyperemia in fed gut than in fasted gut. Our findings suggest that adenosine is a vasodilator metabolite modulating mesenteric reactive hyperemia, especially during enhanced intestinal metabolic activity.

Microcirculatory and motor effects of endogenous nitric oxide in the rat gut.

The aim of the study was to determine the role of endogenous nitric oxide (NO) as the mediator of intestinal blood flow and motility. Experiments were performed on anesthetized rats. Blood flow in the jejunum was determined by Laser-Doppler flowmeter. Motility was monitored on the basis of changes in intrajejunal pressure. Systemic arterial pressure was also recorded. To investigate the potential role of nitric oxide in the regulation of basal intestinal blood flow and motility the NO synthase inhibitor NG-nitro-L-arginine (L-NNA) was given systemically. Intravenous bolus of L-NNA (15 mg/kg) reduced basal intestinal blood flow and increased both intestinal motility and arterial pressure in the dose-dependent manner. To test the specificity of the NO synthase blockade we administered L-arginine alone or in combination with L-NNA. Pretreatment with L-arginine (100.0 mg/kg i.v.) alone had no major influence but when combined with L-NNA it reversed the intestinal circulatory and motor effects of L-NNA. The results of these studies suggest that endogenous NO exerts a tonic relaxatory influence on the smooth muscle of the intestinal vessels and intestinal wall.

Bradykinin-induced mesenteric vasodilation is mediated by B2-subtype receptors and nitric oxide.

We investigated mechanisms mediating bradykinin (BK)-induced anterior mesenteric artery (AMA) vasodilation in anesthetized rats. The velocity of blood flowing (VBF) in the AMA was measured with pulsed Doppler velocimetry, and arterial pressure (BP) was measured with a pressure transducer. Drugs were infused through an intra-aortic catheter positioned proximal to the AMA origin. AMA conductance (C) was calculated from mean VBF/BP and expressed as percent of control C. BK infusion (10-1,000 ng.kg-1.min-1) increased C significantly (Cmax = 201 +/- 18%, ED50 = 100 ng.kg-1.min-1, P < 0.01 for all doses). A B2-subtype receptor antagonist, D-Arg,[Hyp3,Thi5.8,D-Phe7]BK, administered at 10(5) ng.kg-1.min-1 before or during BK infusion, inhibited the vasodilation by 73 +/- 7 and 103 +/- 7%, respectively. A nitric oxide (NO) synthesis inhibitor, NG-nitro-L-arginine, administered at 5.0 mg/kg 15 min before BK, inhibited the hyperemia by 61 +/- 8%. Neither a B1-receptor antagonist nor intrajejunal capsaicin inhibited BK-induced vasodilation. BK-evoked, dose-dependent, mesenteric vasodilation in rats appears to be mediated partly by B2-receptors and endogenous NO generation.

Peptidergic nerves mediate post-nerve stimulation hyperemia in rat gut.

Cessation of perivascular nerve stimulation (NS) elicits a transient increase in intestinal blood flow above the prestimulatory value. This enhancement of blood flow constitutes the phenomenon of post-nerve stimulation hyperemia (PSH). We investigated the involvement of peptidergic sensory nerves in intestinal PSH. In anesthetized rats the velocity of blood flowing through the anterior mesenteric artery (VBF) was measured with a pulsed Doppler velocimeter. PSH was induced by 4 min of postganglionic electrical NS (5 Hz). PSH was abolished by distal periarterial application of tetrodotoxin and intra-arterial lidocaine, which suggests a peripheral sensory nervous mechanism for PSH. The increase in conductance at peak PSH was blocked by pretreatment with the selective, primary afferent neurotoxin capsaicin administered as 1) subcutaneous injection in neonatal life, 2) topical application to periarterial nerves, or 3) injection into the jejunal lumen. In rats pretreated with reserpine, NS evoked a hyperemic response, which was blocked by capsaicin. Treatment with adenosine deaminase inhibited PSH considerably less than capsaicin, suggesting a lesser role for adenosine in PSH. Our findings support the hypothesis that postganglionic NS activates both adrenergic and peptidergic nerves and that the latter release vasodilator peptides in the gut during PSH.

Effects of peptide YY on pancreatic blood flow and oxygen consumption.

Peptide YY (PYY) is a recently discovered polypeptide which has been proposed as physiological inhibitor of pancreatic exocrine secretion. The purpose of this study was to evaluate the effects of exogenous PYY on pancreatic blood flow and oxygen consumption. In anesthetized dogs, the superior pancreatico-duodenal artery blood flow (SPBF), pancreatic microcirculatory blood flow (PBF) and pancreatic oxygen consumption (PVO2) were determined. Control values for SPBF, PVO2 and PBF averaged 43.3 ml/min, 1.8 ml/min, and 57.5 ml/min/100g of tissue, respectively. Following iv injection of PYY at doses of 200 and 400 pmol/kg the values of SPBF decreased by 12 +/- 1% and 22 +/- 2%, respectively. PVO2 was reduced by those doses of PYY by 9 +/- 1 and 17 +/- 3%, respectively. PBF was also reduced by 16 +/- 2 and 34 +/- 2%, respectively after those doses of PYY. Pretreatment with phentolamine reversed the blood flow and PVO2 responses to PYY because SPBF, PVO2 and PBF were significantly increased above the control level. However, after additional pretreatment with propranolol the pancreatic vascular and metabolic responses to PYY were abolished. The above pancreatic responses to PYY were also significantly reduced after acute adrenalectomy. The experimental data indicate that adrenergic pathway is involved in the mechanism of action of PYY on the pancreatic circulation.

Capsaicin-sensitive nerves modulate reactive hyperemia in rat gut.

Reactive hyperemia (RH) is a local, vascular response that occurs following release from mechanical occlusion of an artery, with restoration of intra-arterial pressure. The mechanism of this postocclusion hyperemia in the gut has not been identified, although metabolic, myogenic, and neurogenic mediators of this response have been proposed. The present study was conducted to evaluate a possible modulatory role for sensory innervation of the intestinal vasculature in RH, using acute and chronic treatment with capsaicin applied in different ways. In anesthetized rats, the velocity of flowing blood in the gut was determined continuously with a pulsed Doppler velocimeter, and arterial pressure was determined with a transducer. The increase in calculated intestinal vascular conductance at the height of RH (Ch), the excess volume of blood accumulating during RH, and the duration of the hyperemia were also used to quantify RH after occluding the anterior mesenteric artery for 30, 60, and 120 sec. In the initial control group of rats, the maximal increases in the velocity of flowing blood during RH were 61 +/- 4%, 90 +/- 7%, and 129 +/- 10% of control, conductances were increased to 192 +/- 5%, 222 +/- 12%, and 267 +/- 15% of control, volumes were 3.5 +/- 0.6 ml, 7.2 +/- 0.4 ml, and 16.2 +/- 1.8 ml, and durations of hyperemia were 78 +/- 5 sec, 93 +/- 6 sec, and 178 +/- 7 sec, respectively, after each elapsed period of occlusion. Acute treatment with periarterial capsaicin significantly decreased peak conductances in RH by 15-35% for all occlusions tested and reduced both volume and duration values. Rats treated with capsaicin in neonatal life exhibited reduced Ch values, as did adult rats treated chronically with capsaicin. Both periarterial and intrajejunal treatment with capsaicin decreased the duration of RH. Hexamethonium increased both Ch and the duration of RH and tended to reverse reductions in these parameters caused by capsaicin. These results suggest that sensory innervation of the intestinal vasculature exerts a modulatory influence in the regulation of intestinal RH.

Capsaicin-sensitive nerves mediate inhibitory junction potentials and dilatation in guinea-pig mesenteric artery.

1. The present study examined the effects of repetitive nerve stimulation on membrane potential and on contractile responses to noradrenaline in the guinea-pig inferior mesenteric artery and its distal branches. 2. Repetitive stimulation of perivascular nerves evoked slow inhibitory junction potentials (IJPs) and dilator responses. Individual nerve shocks elicited excitatory junction potentials (EJP)s. 3. Stimulation-evoked IJPs were abolished in the presence of tetrodotoxin (0.3 microM) or a low-Ca2+ (0.5 mM) superfusion solution. 4. The amplitudes and durations of IJPs were dependent on the frequency and duration of repetitive nerve stimulation. Nerve stimulation delivered at 5 Hz for 5 s induced IJPs which had an average amplitude of 2 mV and an average duration of 130 s. When the time interval between successive stimulation periods was less than 4 min, the amplitudes of IJPs were reduced in a time-dependent manner. 5. Stimulation-evoked IJPs were unaffected following endothelium removal. Furthermore, stimulation-evoked IJPs were not affected by atropine (1 microM), indomethacin (20 microM), prazosin (0.5 microM), phentolamine (10 microM), propranolol (0.5 microM) or alpha,beta-methylene ATP (0.2 microM). 6. Pre-treatment of arteries with guanethidine (30 microM) or 6-hydroxydopamine (0.4 mM) abolished stimulation-evoked EJPs but had no effect on stimulation-evoked IJPs. 7. In a similar manner to repetitive nerve stimulation, capsaicin (10 microM) itself induced membrane hyperpolarization and dilatation in mesenteric arteries. Moreover, following application of capsaicin (10 microM), stimulation-evoked IJPs and dilator responses were abolished. 8. EJPs evoked during stimulation-induced IJPs were reduced in amplitude, compared to EJPs evoked under resting conditions. 9. These findings suggest that, in addition to an excitatory sympathetic innervation, mesenteric arteries receive an inhibitory, capsaicin-sensitive innervation which is activated by low-frequency repetitive stimulation.

Comparison of neurotransmission with nerve trunk and transmural field stimulation in guinea-pig mesenteric artery.

1. Intracellular electrical and contractile responses to sympathetic nerve trunk stimulation (NTS) and transmural electrical field stimulation (TMS) were compared in the guinea-pig mesenteric artery in vitro. 2. Step increases in voltage with NTS gave rise to excitatory junctional potentials (EJPs) which reached a plateau amplitude of 5-10 mV, whereas with TMS larger amplitude EJPs and sometimes action potentials were obtained. 3. EJPs of equal amplitude (1-7 mV) elicited with TMS and NTS had the same rise time, duration and decay half-time. 4. Slow depolarization obtained with repetitive stimulation was significantly greater in amplitude with TMS than with NTS. 5. Equal amplitude EJPs were obtained throughout the preparation with NTS. With TMS, the amplitude of responses declined substantially with distance from the stimulating electrodes. 6. Tetrodotoxin (TTX) completely blocked EJPs, slow depolarization and contraction with NTS; however, with TMS a TTX-resistant component was observed. The TTX-resistant response to TMS was abolished in the presence of a low-Ca2+ superfusion solution but was not affected by endothelium removal. 7. Phentolamine or prazosin abolished slow depolarization but not EJPs with NTS or TMS. Prazosin abolished contraction with NTS and reduced but did not abolish contraction with TMS. 8. alpha, beta-Methylene ATP abolished EJPs with NTS, whereas with TMS only EJPs obtained with low stimulus intensities were abolished. alpha, beta-Methylene ATP did not block contraction with either NTS or TMS. 9. Combined TTX, prazosin and alpha, beta-methylene ATP abolished EJPs initiated with TMS at all but the highest stimulus intensities (12-20 V, 0.3 ms duration). 10. It is concluded that responses obtained with NTS can be reliably attributed to the release of transmitter by the conduction of action potentials in paravascular nerves, whereas activation by TMS is a more complex phenomenon dependent upon stimulus strength and probably involving multiple forms of activation.

Adrenergic modulation of reactive hyperemia in rat gut.

The hypothesis was tested that peripheral, adrenergic nerves modulate reactive hyperemia (RH) in the intestinal circulation. In anesthetized rats, anterior mesenteric arterial occlusion for 30-120 s elicited subsequent RH responses, including 63-118% increases in the velocity of arterial blood flow, even greater increases in conductance, and durations of 64-139 s. The longer the period of arterial occlusion, the greater the magnitude of RH. Electrical stimulation of postganglionic, sympathetic nerves reduced RH responses in a frequency-dependent manner. RH responses were enhanced by pretreatment with hexamethonium and phenoxybenzamine and were diminished by pretreatment with propranolol. Propranolol also prevented the enhanced RH responses caused by hexamethonium and phenoxybenzamine. Reserpine prevented the enhanced RH responses to hexamethonium, but bilateral adrenalectomy did not. These findings support the hypothesis that peripheral sympathetic nerves modulate RH in rat gut, with alpha-adrenergic receptors restricting and beta-adrenergic receptors enhancing the hyperemia.

Capsaicin-sensitive nerves modulate resting blood flow and vascular tone in rat gut.

Acute and chronic treatments with capsaicin were used to evaluate the role of afferent neurons in the regulation of intestinal blood flow. Experiments were performed on anesthetized rats, in which mean intestinal blood flow was determined with a pulsed Doppler flowmeter, mean systemic arterial pressure was determined with a transducer, and intestinal vascular conductance (C) was calculated from these measurements. Acute administration of periarterial capsaicin (0.5 mg) induced biphasic intestinal vascular responses. An early hyperemic response occurred with a maximal increase in blood flow of 31% at 5 min, followed by a decrease in blood flow of 17% at 30 min. Arterial pressure was decreased by the application of capsaicin, initially by 10%. There was an early increase of 49% in conductance, followed by a 15% decrease, compared with control values. When 1 or 4 mg capsaicin was instilled into the lumen of the jejunum there was a response pattern similar to that observed after periarterial application of capsaicin. Intrajejunal capsaicin (4 mg) increased blood flow by 51%, followed by a decrease of 16%. Mean mesenteric artery conductance was increased by 32% initially and subsequently was decreased by 21%, in response to acute intrajejunal administration of capsaicin. Both mean blood flow and conductance were increased (44% and 76%, respectively) in adult rats chronically pretreated with capsaicin (170 mg total dose) when compared with vehicle-treated controls. However, in rats pretreated neonatally with capsaicin (50 mg/kg) and allowed to mature, basal flood flow was lower than in control animals but C was not different from control littermates.(ABSTRACT TRUNCATED AT 250 WORDS)

[Role of adenosine in functional and reactive intestinal hyperemia]. / Rola adenozyny w czynnym i reaktywnym przekrwieniu jelit.

This study was designed to evaluate the role of adenosine and adenosine receptors in the reactive (RH) and functional hyperemia (FH) in rat gut. Experiments were performed on anesthetized rats. Mesenteric blood flow was measured with a pulsed Doppler flowmeter. We also determined the duration of reactive hyperemia, excess volume of blood flow above control value and maximal increase in mesenteric vascular conductance during both hyperemic responses. Data were collected following release from occlusions lasting 30, 60 and 120 sec. Functional hyperemia was induced by perfusion of the gut with a solution. Studied parameters were obtain before and after adenosine deaminase (ADA) and two adenosine receptor antagonists: 8-phenyltheophylline (8-PT) and 1.3-dipropyl-7-methyl-xanthine (DPMX). In fasted rats ADA and 8-PT reduced of RH after each period of occlusion and DPMX was ineffective in reducing any parameter of RH. In fed rats control mesenteric blood flow was increased. ADA, 8-PT, and DPMX were more effective inhibitors of RH and FH. Above findings suggest that adenosine play a role in the modulation of RH and FH acting on A2 subtype receptors.

Sensory nerves mediate neurogenic escape in rat gut.

We investigated the involvement of primary sensory nerves in intestinal autoregulatory escape induced by postganglionic nerve stimulation (NS) in anesthetized rats. Anterior mesenteric artery (AMA) blood flow velocity (BF) was measured with a pulsed Doppler flowmeter. Periarterial NS elicited an abrupt fall in BF, which was followed by a recovery in BF toward the basal value, despite sustained NS. This recovery from NS constituted the neurogenic escape phenomenon. Vasoconstrictor responses to NS were abolished by periarterial application of tetrodotoxin. Acute, surgical interruption of proximal periarterial nerves had no effect on BF responses to distal NS, suggesting a peripheral rather than a central nervous mechanism for the escape phenomenon. Escape from NS-induced vasoconstriction was significantly inhibited by prior administration of the selective sensory neurotoxin capsaicin as either subcutaneous injection in neonatal life, acute application to periarterial nerves, or acute injection into the jejunal lumen. In rats pretreated 24 h with reserpine, NS provoked a vasodilator response that was inhibited by intrajejunal capsaicin. Increases in arterial blood pressure (BP) and heart rate observed during NS were blocked by periarterial (but not intrajejunal) application of capsaicin. Transmural electrical field stimulation elicited significantly greater nerve-induced contractions in AMA rings from control rats. Our findings support the hypothesis that postganglionic NS activates both vasoconstrictor sympathetic nerve branches and vasodilator afferent C-fibers. The latter nerves release vasodilator peptides in the periphery during continuous low frequency NS that appear to be essential for autoregulatory escaped in our model.

Comparison of the frequency dependence of venous and arterial responses to sympathetic nerve stimulation in guinea-pigs.

1. Intracellular potentials and measurements of contractions were recorded in adjacent veins and arteries in the colonic mesentery of the guinea-pig in vitro during stimulation of post-ganglionic nerve trunks. 2. Repetitive stimulation (0.5-5 Hz) of lumbar colonic nerve trunks produced frequency-dependent slow depolarizations in all venous and in 92% of arterial smooth muscle cells. Excitatory junction potentials were observed for each nerve shock in arteries, but not in veins. 3. Low-frequency stimulations produced slow depolarizations of greater amplitude and longer duration in veins than in arteries. The frequencies at which half-maximal depolarizations and contractions occurred were always lower for veins than for arteries. 4. The alpha 1-adrenergic antagonist prazosin (5 X 10(-7) M) reduced the mean arterial slow depolarizations by 82% and reduced mean venous slow depolarizations by 58% for 5 Hz stimulations. Arterial contractions were completely inhibited by prazosin but venous contractions were incompletely reduced in a frequency-dependent manner. 5. These findings suggest that functional differences in activation between mesenteric veins and arteries during sympathetic stimulation are a result of differences in neuromuscular transmission.