Role of alpha-adrenoceptors and prostacyclin in the enhanced adrenergic reactivity of goat cerebral arteries after ischemia-reperfusion.

To analyze ischemia-reperfusion effects on the cerebrovascular adrenergic response, the left middle cerebral artery (MCA) of anesthetized goats was occluded for 120 min and reperfused for 60 min. Isolated segments from the left (ischemic) and right (control) MCA exhibited isometric constriction in response to noradrenaline (10(-8)-10(-4)M, in the presence of beta-adrenoceptors blockade), phenylephrine (alpha(1)-adrenoceptors agonist, 10(-8)-10(-4)M), B-HT-920 (alpha(2)-adrenoceptors agonist, 10(-7) - 3 x 10(-3)M) or tyramine (indirect sympatheticomimetic amine, 10(-8)-10(-4)M), but this constriction was greater in ischemic arteries. The cyclooxygenase (COX) inhibitor meclofenamate (10(-5)M) augmented the response to noradrenaline only in control arteries. The prostacyclin (PGI(2)) synthesis inhibitor tranylcypromine (TCP, 10(-5)M) increased the response to noradrenaline in control arteries and reduced it in ischemic arteries. The thromboxane A(2) (TXA(2)) synthase inhibitor furegrelate (10(-6)M) did not modify the noradrenaline effect in both types of arteries, whereas the TXA(2) receptor antagonist SQ 29 548 (10(-5)M) and the COX-2 inhibitor NS-398 (10(-6)M) decreased the response to noradrenaline only in ischemic arteries. PGI(2) caused a small relaxation in control arteries and a small contraction in ischemic arteries. alpha-Adrenoceptors and COX-2 protein expression and the metabolite of PGI(2) were augmented in ischemic arteries. Therefore, ischemia-reperfusion may increase the cerebrovascular responsiveness to noradrenaline, through upregulation of alpha-adrenoceptors and increased COX-2-derived PGI(2) exerting a vasoconstrictor action. After ischemia-reperfusion, noradrenaline might increase PGI(2) production thus contributing to adrenergic vasoconstriction and/or PGI(2) would potentiate the noradrenaline effects.

Effects of endothelin-1 on the relaxation of rat coronary arteries.

To analyze the effects of endothelin-1 on the b-adrenergic response of the coronary circulation, 2-mm-long segments of coronary arteries from rats were prepared for isometric tension recording in organ baths. The relaxation to isoproterenol (3 x 10(-8) M), field electrical stimulation (4 Hz, 0.1-millisecond duration, 10 seconds), acetylcholine (3 x 10(-8) M), and sodium nitroprusside (10(-9) M) was recorded in arteries precontracted with U46619 (10(-7) to 5 x 10(-7) M) before and after treatment with endothelin-1 (3 3 10210 and 1029 M). The relaxation to isoproterenol was increased by treatment with endothelin-1 and with the endothelin ET(B) antagonist BQ788 (10(-6) M) but not with the endothelin ET(A) antagonist BQ123 (10(-6) M) or with the blocker of protein kinase C chelerythrine (10(-5) M). In the presence of BQ788, BQ123, or chelerythrine, endothelin-1 did not modify the relaxation to isoproterenol. Treatment with endothelin-1 did not modify the relaxation to electrical stimulation, acetylcholine, or sodium nitroprusside. These results suggest that endothelin-1 may potentiate coronary beta-adrenergic vasodilatation, at least in part due to stimulation of endothelin ET(A) receptors and activation of protein kinase C.

Role of angiotensin II in the response to endothelin-1 of goat cerebral arteries after ischemia-reperfusion.

As angiotensin II may underlie the deleterious effects of some vascular diseases, we have examined the role of this peptide on the cerbrovascular endothelin-1 action after ischemia-reperfusion. In anesthetized goats, 1 hour-occlusion followed by 1 hour-reperfusion of the left middle cerebral artery (MCA) was induced, and then segments 3-mm in length from branches of the right MCA (control) and the left MCA (ischemic) were obtained for isometric tension recording. Endothelin-1 (10(-11)-10(-7) M) produced a contraction that was higher in ischemic than in control arteries, and in control but not in ischemic arteries this contraction was potentiated by angiotensin II (10(-7) M). Losartan (3 x 10(-6) M), antagonist of AT1 receptors, did not affect the response to endothelin-1 in control arteries, but reduced it both in ischemic arteries and angiotensin II-treated control arteries. PD123,319 (3 x 10(-6) M), antagonist of AT2 receptors, or the inhibitor of nitric oxide synthesis L-NAME (10(-4) M) did not alter the arterial effects of endothelin-1. Therefore, angiotensin II may potentiate the constriction to endothelin-1 in normal cerebral arteries by activating AT1 receptors. The observed cerebrovascular increased response to endothelin-1 after ischemia-reperfusion might be related in part to activation of AT1 receptors under this condition.

Coronary response to diadenosine pentaphosphate after ischaemia-reperfusion in the isolated rat heart.

AIMS: Diadenosine polyphosphates are vasoactive mediators that may be released from platelet granules and which may be present at higher concentrations during coronary ischaemia-reperfusion. The objective of this study was to analyse their effects in such conditions. METHODS AND RESULTS: Rat hearts were perfused in a Langendorff preparation and the response to diadenosine pentaphosphate (Ap5A, 10(-7)-10(-5) M) was recorded. In control hearts, Ap5A produced a small, transient coronary vasoconstriction followed by marked vasodilatation, as well as a reduction in the left ventricular developed pressure dP/dt and heart rate, both at the basal coronary resting tone or after pre-contracting coronary arteries with 9,11-dideoxy-11alpha, 9alpha-epoxymethanoprostaglandin F2alpha (U46619). After ischaemia-reperfusion, the vasoconstriction in response to Ap5A was augmented and vasodilatation diminished, both in hearts with basal or increased vascular tone. The pyridoxal derivative P(2) purinoceptor antagonist, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS, 3 x 10(-6) M), inhibited this vasoconstriction, while the antagonist of purinergic P(2Y) receptors, Reactive Blue 2 (2 x 10(-6) M), inhibited the vasodilatation, both before and after ischaemia-reperfusion. The antagonist of nitric oxide synthesis N-omega-nitro-L- arginine methyl ester (L-NAME, 10(-4) M) did not modify the response to Ap5A, whereas the cyclooxygenase inhibitor, meclofenamate (2 x 10(-6) M), reduced contraction and increased the relaxation in response to Ap5A after ischaemia-reperfusion but not under control conditions. CONCLUSION: Ischaemia-reperfusion reduces the vasodilatory response to Ap5A and increases the vasoconstriction provoked due to a reduced influence of purinergic P(2Y) receptors and/or to the production of vasoconstrictor prostanoids.

Response to endothelin-1 in arteries from human colorectal tumours: role of endothelin receptors.

To examine the reaction of tumour arteries to endothelin-1, we obtained arteries supplying blood flow to colorectal tumours from patients, as well as mesenteric arteries supplying the normal colon tissue from the same patients and mesenteric arteries from patients without a colorectal tumour pathology. The contraction in response to endothelin-1 and the relaxation produced by bradykinin was recorded in each of these arteries. Accordingly, the sensitivity to endothelin-1 but not the maximal response, was higher in the arteries supplying colorectal tumours than in mesenteric arteries supplying normal colon or in mesenteric arteries from patients with no tumour pathology. The contraction produced by endothelin-1 was not modified by exposure to L-NAME or meclofenamate in arteries supplying both the tumour and the normal colon. The endothelin ET(A) andET(B) receptors were expressed similarly in arteries supplying the tumour or normal colon. However, the antagonist of the endothelin ET(B) receptors BQ788 (10(-6) M) decreased the contractions in the arteries supplying the tumour but not in those supplying the normal colon. By contrast, the antagonist of endothelin ET(A) receptors BQ123 (10(-6) M) reduced the contraction equally in both these types of arteries. Likewise, in arteries precontracted with U46619, the relaxation in response to bradykinin was similar in all three types of arteries. Together, these results suggest that the arteries supplying human colorectal tumours are more sensitive to endothelin-1, which could be due to the enhanced activity of endothelin ET(B) receptors in the absence of any change in the modulatory effect of nitric oxide or prostanoids in the arterial response to this peptide.

Endothelin-1 potentiation of coronary artery contraction after ischemia-reperfusion.

Hearts from Sprague-Dawley rats were perfused at constant flow and then exposed to 30 min global zero-flow ischemia followed by 15 min reperfusion. After ischemia-reperfusion, coronary arteries were dissected from the heart and segments 2 mm long were prepared for isometric tension recording in organ baths. Stimulation of the arteries with 5-hydroxytryptamine (10(-6) M) produced contraction, which was potentiated by treatment with endothelin-1 (3x10(-10); 10(-9) M). This potentiation was lower in the arteries from hearts after ischemia-reperfusion (for 3x10(-10) M, 15+/-5%; P>0.05; for 10(-9) M, 37+/-7%, P<0.01, n=5) than after control (for 3x10(-10) M, 34+/-4%; P<0.01; for 10(-9) M, 50+/-6%, P<0.01, n=5), and the potentiation was reduced by the inhibitor of nitric oxide synthesis l-NAME (10(-4) M), the antagonist of endothelin ET(A) receptors BQ123 (10(-6) M) and the antagonist of endothelin ET(B) receptors BQ788 (10(-6) M), but not by the cyclooxygenase inhibitor meclofenamate (10(-5) M). These results suggest that endothelin-1 at low concentrations potentiates coronary vasoconstriction, and this effect is reduced after ischemia-reperfusion, mediated by endothelin ET(A) and ET(B) receptors and dependent on nitric oxide release.

Adrenergic response of splanchnic arteries from cirrhotic patients: role of nitric oxide, prostanoids, and reactive oxygen species.

Peripheral and splanchnic vasodilatation in cirrhotic patients has been related to hyporesponsiveness to vasoconstrictors, but studies to examine the vascular adrenergic response provide contradictory results. Hepatic arteries from cirrhotic patients undergoing liver transplantation and mesenteric arteries from liver donors were obtained. Segments 3 mm long from these arteries were mounted in organ baths for testing isometric adrenergic response. The concentration-dependent contraction to noradrenaline (10(-8) to 10(-4) M) was similar in hepatic and mesenteric arteries, and prazosin (alpha 1-adrenergic antagonist, 10(-6) M), but not yohimbine (alpha 2-adrenergic antagonist, 10(-6) M), produced a rightward parallel displacement of this contraction in both types of arteries. Phenylephrine (alpha 1-adrenergic agonist, 10(-8) to 10(-4) M) and clonidine (alpha 2-adrenergic agonist, 10(-8) to 10(-4) M) also produced concentration-dependent contractions that were comparable in hepatic and mesenteric arteries. The inhibitor of cyclooxygenase meclofenamate (10(-5) M), but not the inhibitor of nitric oxide synthesis N(w)-nitro-l-arginine methyl ester (l-NAME, 10(-4) M), potentiated the response to noradrenaline in hepatic arteries; neither inhibitor affected the response to noradrenaline in mesenteric arteries. Diphenyleneiodonium (DPI; 5 x 10(-6) M), but neither catalase (1000 U/ml) nor tiron (10(-4) M), decreased the maximal contraction for noradrenaline similarly in hepatic and mesenteric arteries. Therefore, it is suggested that, in splanchnic arteries from cirrhotic patients, the adrenergic response and the relative contribution of alpha 1- and alpha 2-adrenoceptors in this response is preserved, and prostanoids, but not nitric oxide, may blunt that response. Products dependent on NAD(P)H oxidase might contribute to the adrenergic response in splanchnic arteries from control and cirrhotic patients.

Apelin effects in human splanchnic arteries. Role of nitric oxide and prostanoids.

Apelin effects were examined in human splanchnic arteries from liver donors (normal arteries) and from liver recipients. Segments 3 mm long were obtained from mesenteric arteries taken from liver donors (normal arteries), and from hepatic arteries taken from cirrhotic patients undergoing liver transplantation (liver recipients), and the segments were mounted in organ baths for isometric tension recording. In arteries under resting conditions, apelin (10(-10)-10(-6) M) caused no effect in any of the arteries tested. In arteries precontracted with the thromboxane A(2) analogue U46619 (10(-7)-10(-6) M), apelin (10(-10)-10(-6) M) produced concentration-dependent relaxation that was lower in hepatic than in mesenteric arteries, whereas sodium nitroprusside (10(-8)-10(-4) M) produced a similar relaxation in both types of arteries. The inhibitor of nitric oxide synthesis N(w)-nitro-L-arginine methyl ester (L-NAME, 10(-4) M) diminished the relaxation to apelin in mesenteric but not in hepatic arteries. The inhibitor of cyclooxygenase meclofenamate (10(-5) M) did not affect the relaxation provoked by apelin in both types of arteries. Therefore, apelin may produce relaxation in normal human splanchnic arteries, and this relaxation may be mediated in part by nitric oxide without involvement of prostanoids. This relaxation as well as the role of nitric oxide may be decreased in splanchnic arteries from cirrhotic patients.

Endothelium-dependent relaxation of isolated splanchnic arteries from cirrhotic patients: Role of reactive oxygen species.

AIM: To examine the endothelium-dependent relaxation of splanchnic arteries during cirrhosis as well as the role of reactive oxygen species in this relaxation using hepatic arteries from cirrhotic patients undergoing liver transplantation and mesenteric arteries from liver donors. METHODS: Arterial segments 3 mm long were mounted in organ baths for isometric tension recording and precontracted with the thromboxane A(2) analog U46619 (10(-7)-10(-6) M). RESULTS: The relaxation to acetylcholine (10(-8)-10(-4) M), but not to sodium nitroprusside (10(-8)-10(-4) M) was lower in hepatic arteries. The inhibitor of nitric oxide synthesis, N(omega)-nitro-l-arginine methyl ester (l-NAME, 10(-4) M), the inhibitor of cyclooxygenase, meclofenamate (10(-5) M), or l-NAME (10(-4) M) + meclofenamate (10(-5) M) diminished the relaxation to acetylcholine only in mesenteric arteries. l-NAME (10(-4) M) + meclofenamate (10(-5) M) combined with charybdotoxin (10(-7) M) + apamine (10(-6) M) inhibited the relaxation toacetylcholine in both types of arteries, and this inhibition was greater than with l-NAME + meclofenamate. The scavenger of hydrogen peroxide, catalase (1000 U/mL), the superoxide dismutase mimetic, tiron (10(-2) M) or the inhibitor of NAD(P)H oxidase, diphenyleneiodonium (5 x 10(-6) M), but not the inhibitor of superoxide dismutase, diethyldithiocarbamate (10(-3) M) potentiated the acetylcholine-induced relaxation only in hepatic arteries. l-NAME did inhibit the relaxation to acetylcholine in hepatic arteries pretreated with catalase or tiron. CONCLUSIONS: Cirrhosis may decrease endothelial release and/or bioavailability of nitric oxide and prostacyclin in splanchnic arteries, which might be caused partly by increased production of reactive oxygen species.

Goat cerebrovascular reactivity to ADP after ischemia-reperfusion. Role of nitric oxide, prostanoids and reactive oxygen species.

To analyze the cerebrovascular effects of ischemia-reperfusion, cerebrovascular reactivity to ADP was studied after inducing 60-min occlusion followed by 60-min reperfusion of the left middle cerebral artery (MCA) in anesthetized goats. In 12 goats, at the end of reperfusion, left MCA resistance was decreased by 19%, and reactive hyperemia to 5- and 10-s occlusions as well as the cerebral vasodilatation to ADP (0.03-0.3 microg) but not to sodium nitroprusside (0.3-3 microg) was decreased. In 28 animals, killed at the end of reperfusion, segments 3-mm long were obtained from the left (ischemic) and right (control) MCA, prepared for isometric tension recording, and precontracted with the thromboxane A2 analogue U46619. The relaxation to ADP (10(-8) to 10(-5) M) but not to sodium nitroprusside (10(-8) to 10(-4) M) was lower in ischemic arteries. L-NAME (inhibitor of nitric oxide synthesis, 10(-4) M), charybdotoxin (10(-7) M)+apamin (10(-6) M) (blockers of KCa), or catalase (1000 U/ml) reduced the relaxation to ADP only in control arteries. Charybdotoxin+apamin further augmented the L-NAME-induced reduction in the relaxation to ADP in control arteries. The inhibitor of cyclooxygenase meclofenamate (10(-5) M) increased the relaxation to ADP only in ischemic arteries. The superoxide dismutase mimetic tiron (10(-2) M) increased the ADP-induced relaxation only in ischemic arteries. Therefore, it is suggested that ischemia-reperfusion produces cerebrovascular endothelial dysfunction, which may be associated with decreased nitric oxide bioavailability, decreased release of an EDHF, and increased production of vasoconstrictor prostanoids. All these alterations may be related in part with an increased production of superoxide anion.