Functional characterization of the muscarinic receptors involved in endothelium-dependent relaxation in isolated canine uterine artery.

Acetylcholine interacts with endothelial muscarinic receptors releasing nitric oxide and causing vasodilatation. To identify the receptor subtype responsible for acetylcholine-induced relaxation in canine uterine artery, the usual organ bath method for in vitro investigation on isolated blood vessels was applied. Using a range of muscarinic receptor antagonists such as atropine (nonselective), pirenzepine (M(1)-selective), methoctramine (M(2)-selective) and p-fluoro-hexahydro-sila-difenidol (p-FHHSiD) (M(1)/M(3)) and determining pA2 value of those antagonists through Shild analysis, we aimed at establishing a precise receptor mechanism underlying acetylcholine-induced relaxation in isolated canine uterine artery. The relaxation of uterine arterial rings in response to acetylcholine in the presence or absence of selective muscarinic receptors antagonists was calculated using concentration response curves. Acetylcholine induced concentration-dependent and endothelium-dependent relaxation of arterial rings precontracted with phenylephrine (pEC(50) = 6.90 +/- 0.02). Muscarinic receptors antagonists atropine, pirenzepine, methoctramine and p-FHHSiD competitively antagonized the response to acetylcholine and obtained pA(2) values were 9.91 +/- 0.06, 6.60 +/- 0.04, 6.21 +/- 0.08 and 8.05 +/- 0.1, respectively. This study showed that acetylcholine induced endothelium-dependent relaxation of canine uterine artery by stimulation of muscarinic receptors localized on the endothelial cells. On the basis of differential antagonist affinity, we suggest that the muscarinic receptors involved in the acetylcholine-induced relaxation of canine uterine artery are predominantly of M(3) subtype.

Responses of the human submandibular artery to ACh and VIP.

Endothelial vasodilatory substances may play a central role in the local regulation of vascular tone. We hypothesized that these substances can mediate endothelium-dependent vasodilatory responses to acetylcholine (ACh) and vasoactive intestinal peptide (VIP) in the human submandibular artery. We evaluated the contributions of endothelial vasodilatory substances to vessel relaxation in response to ACh and VIP, using different inhibitors of endothelial vasodilation, the nitric oxide synthase inhibitor, the cyclo-oxygenase inhibitor, indomethacin, the potassium channel blocker, and 4-aminopyridine. ACh and VIP caused an endothelium- and concentration-dependent relaxation in this artery. ACh relaxation was completely blocked after the concomitant addition of N(G)-nitro-L-arginine and indomethacin. The vasorelaxant effect of ACh was not influenced by 4-aminopyridine. VIP relaxation was almost completely abolished by 4-aminopyridine, and was partly inhibited by N(G)-nitro-L-arginine, but was not affected by indomethacin. Thus, in the human submandibular artery, ACh and VIP produced endothelium-dependent vasodilation with different underlying mechanisms: release of nitric oxide (NO) and cyclo-oxygenase products for ACh, and release of NO and endothelium-derived hyperpolarizing factor for VIP.

The effects of acute and chronic lithium treatment on rat submandibular salivation.

OBJECTIVE: Acute and chronic actions of lithium on salivation induced by agonists associated with receptor-linked hydrolysis of membrane inositol phospholipids (carbachol and phenylephrine) and by agonist linked to activation of adenylate cyclase (isoproterenol) were investigated. MATERIAL AND METHODS: In anaesthetized rats, submandibular salivation induced by intravenous injection of carbachol, phenylephrine and isoproterenol, was measured and expressed as volume of fluid (microl) elicited per 100 mg wet weight of each gland per minute. The experiments were repeated after acute and chronic treatment of lithium (7 mg kg(-1)). The results were analysed with unpaired t-test. RESULTS: Chronic, but not acute lithium treatment significantly decreases carbachol- and phenylephrine-induced salivation while isoproterenol-induced salivation was not changed neither after acute nor after chronic administration of lithium. CONCLUSION: The results suggest that hyposalivation during chronic lithium therapy could be mediated by alterations in the phosphatidylinositol cycle and a consequent lack of inositol after agonist stimulation.

The relaxant effect of vasoactive intestinal polypeptide in the isolated canine uterine artery: the role of endothelium.

The purpose of this study was to examine the effect of vasoactive intestinal polypeptide (VIP) on the uterine artery obtained from non-pregnant dogs. VIP (3 x 10(-9)-3 x 10(-7) M) induced concentration-dependent relaxation in canine uterine arteries with intact endothelium, pre-contracted with 10(-5) M phenylephrine (pEC(50) = 7.52 +/- 0.02, maximal response was 82.19 +/- 2.15%, n = 36). The administration of the cyclooxygenase inhibitor indomethacin (10(-5) M) or 4-aminopyridine (4-AP), a blocker of potassium channels (10(-5) M), did not modify the relaxation induced by VIP. Contrary to this, N(G)-nitro-L-arginine (L-NOARG) (10(-5) M) inhibited relaxation is evoked by VIP. Indomethacin applied with L-NOARG did not provoke further inhibition of VIP-induced relaxation. In the presence of both L-NOARG and L-NOARG + indomethacin, 4-AP led to the further inhibition of VIP-induced relaxation of canine uterine artery. It is concluded that VIP induces endothelium-dependent relaxation of uterine arteries of non-pregnant dogs, which can be entirely explained by the production of nitric oxide (NO) from the endothelial cells. We proposed that when NO synthesis is inhibited, VIP induces further relaxation, independent of the edothelium-derived relaxing factors, probably through activation of K(+) channels.

Endothelium-dependent relaxation of canine uterine artery in response to acetylcholine: the possible involvement of alternative pathways.

The effect of acetylcholine on the isolated, pre-contracted, uterine artery of non-pregnant dog was investigated. Acetylcholine-induced concentration-dependent relaxation of isolated canine uterine artery with endothelium (pEC50 = 6.48 +/-0.01, n = 37) and was without effect on arterial segments denuded of endothelium. Indomethacin, 4-aminopyridine (10-5 m) and pre-contraction with K+-rich Krebs-Ringer bicarbonate solution had no effect on acetylcholine-induced relaxation. NG-nitro-l-arginine (l-NOARG) (10-5 m) inhibited relaxation evoked by acetylcholine. Indomethacin applied with l-NOARG led to further inhibition of acetylcholine-induced relaxation. In the presence of both l-NOARG and indomethacin, 4-aminopiridine did not provoke further inhibition of acetylcholine-induced relaxation of canine uterine artery. It is concluded that the acetylcholine-induced relaxation of canine uterine artery is probably mediated by endothelial production of nitric oxide (NO). However, if NO-synthase is inhibited, acetylcholine-induced vasorelaxation may be, in part, mediated through activation of cyclooxygenase pathway.

Acetylcholine-induced contractions in the perforating branch of the human internal mammary artery: protective role of the vascular endothelium.

The effect of acetylcholine (ACh) on the isolated, nonprecontracted perforating branch of the human internal mammary artery (HIMA) was investigated. ACh induced concentration-dependent contractions of nonprecontracted rings with denuded endothelium (pEC(50) = 6.72 +/- 0.02, E(max) = 88.8% of contractions induced by phenylephrine, 10(-5) mol/l) and was without effect on arterial segments with intact endothelium. An inhibitor of nitric oxide synthase, N(G)-monomethyl-L-arginine (L-NMMA), or indometacin, a cyclooxygenase inhibitor, had no effect on acetylcholine-induced contractions of rings of the perforating branch of HIMA with denuded endothelium (pEC(50) = 6.76 +/- 0.03 and 6.62 +/- 0.05, respectively). In the presence of indometacin, ACh did not evoke contractions of arterial segments with intact endothelium. In contrast, in the same type of preparations ACh induced contractions in the presence of L-NMMA (E(max) = 34%). The muscarinic receptor antagonists atropine (no selectivity), pirenzepine (M(1)), methoctramine (M(2)), and p-fluoro-hexahydro-sila-difenidol (M(1)/M(3)) competitively antagonized the response to ACh. The pA(2) values were 9.60 +/- 0.10, 6.99 +/- 0.02, 6.37 +/- 0.17, and 8.02 +/- 0.06, respectively. In conclusion, the results obtained indicate that secretion of nitric oxide from vascular endothelium may protect the perforating branch of HIMA against the contractile effects of ACh. On the basis of differential antagonist affinity, it can be suggested that the muscarinic receptors involved in the ACh-induced contractions of the isolated perforating branch of the HIMA are predominantly of the M(3) subtype.

Muscarinic receptor subtypes mediating vasorelaxation of the perforating branch of the human internal mammary artery.

The effect of acetylcholine (ACh) on the isolated perforating branch of the human internal mammary artery (HIMA) was investigated. ACh induced concentration- and endothelium-dependent relaxation of arterial rings precontracted with phenylephrine (pEC(50) = 6.93 +/- 0.01). The muscarinic receptor antagonist atropine (no selectivity), pirenzepine (M(1)), methoctramine (M(2)), and p-fluoro-hexahydro-siladifenidol (M(1)/M(3)) competitively antagonized the response to ACh. The pA(2) values were 9.81 +/- 0.15, 7.74 +/- 0.08, 6.27 +/- 0.08, and 7.88 +/- 0.04, respectively. In conclusion, this study has shown that ACh induced an endothelium-dependent relaxation of the perforating branch of the HIMA by stimulation of muscarinic receptors on the endothelial cells. On the basis of differential antagonist affinity, we suggest that the muscarinic receptors involved in the ACh-induced relaxation of the isolated perforating branch of HIMA are predominantly of M(1) subtype.

Endothelium-dependent relaxation in perforating branch of human internal mammary artery.

The purpose of the present study was to examine the effect of acetylcholine on perforating branch of the human internal mammary artery (HIMA). Acetylcholine (10(-9)-10(-5)M) induced concentration- and endothelium-dependent relaxation (pEC(50)=7.54+/-0.03, maximal response was 98+/-1.3%) of the precontracted arterial segments. Indomethacin, 4-aminopyridine (10(-5)M) and precontraction with K(+)-rich Krebs-Ringer-bicarbonate solution had no effect on acethylcholine-induced relaxation. N(G)-monomethyl-L-arginine (L-NMMA) (10(-5)M) inhibited relaxation evoked by acetylcholine. Indomethacin applied together with L-NMMA lead to further inhibition of acethylcholine-induced relaxation. Even in the presence of both L-NMMA and indomethacin, 4-aminopyridine had no provoked further inhibition of acetylcholine-induced relaxation of perforating branch of HIMA. It was concluded that the acethylcholine-induced relaxation of isolated perforating branch of HIMA is probably mediated via endothelial production of nitric oxide. However, when NO-synthase is inhibited, acetylcholine-induced vasorelaxation may be, in part, mediated through activation of cyclooxygenase pathway and consequent production and release of prostacyclin or some other cyclooxygenase products.

Pregnancy is associated with altered response to neuropeptide Y in uterine artery.

Pregnancy is associated with a significant increase in uterine blood flow which contributes to optimal fetal development. Although neuropeptide Y (NPY) is considered to be an important regulator of uterine blood flow, it is not known whether: (i) products from the vascular endothelium modulate NPY action in the uterine artery; (ii) pregnancy changes the responsiveness of the uterine artery to NPY, or (iii) NPY interacts with noradrenaline and acetylcholine on the uterine artery, with pregnancy regulating this possible interaction. In the present study, NPY induced a concentration-dependent contraction of guinea pig uterine arterial rings both intact and denuded of endothelium. Pregnancy significantly decreased the potency of NPY to contract uterine artery with and without endothelium. In all preparations, addition of N(G)-monomethyl-l-arginine acetate (l-NMMA), indomethacin and diethylcarbamazine did not modify the effect of NPY. In the presence of NPY concentration-response curves for acetylcholine and noradrenaline were significantly shifted to the right and left respectively. This effect of NPY was independent of endothelial condition or pregnancy status. The receptor reserve (K(A)/EC(50)) for acetylcholine was decreased and for noradrenaline was increased in the presence of NPY, although no changes in the dissociation constants of the neurotransmitter-receptor complexes were observed. Thus, this study has shown that: (i) NPY induces contraction of guinea pig uterine arteries acting on receptors localized in smooth muscle; (ii) pregnancy alters the response of guinea pig uterine arteries to NPY in such a way as to promote vasorelaxation, and (iii) NPY modulates the effect of neurotransmitters on guinea pig uterine arteries, but pregnancy is not associated with the changes at the level of NPY-neurotransmitter interaction.

Pregnancy does not alter the response of uterine arteries to vasoactive intestinal polypeptide.

In order to provide sufficient nutrients for fetal development, pregnancy is associated with a significant increase in uterine blood flow. Although vasoactive intestinal polypeptide (VIP) is considered to be an important regulator of uterine blood flow it is not known whether endothelium-derived relaxing factors contribute to VIP action in the uterine artery, whether pregnancy alters the effect of VIP in the uterine artery and/or whether VIP interacts with noradrenaline and acetylcholine on the uterine artery and whether pregnancy regulates this possible interaction. In the present study, VIP induced a concentration-dependent relaxation of guinea pig uterine arterial rings, both intact and denuded of endothelium. Pregnancy did not alter the relaxation of uterine artery in response to VIP. In all preparations, addition of N(G)-monomethyl-L-arginine (L-NMMA), indomethacin and diethylcarbamazine did not modify the effect of VIP in uterine arteries. The VIP-receptor complex dissociation constant did not differ significantly between studied vessels, and in all experimental groups the relationship between receptor occupancy and the response was linear, with the receptor reserve (K(A)/EC(50)) close to unity. VIP did not modulate acetylcholine-induced relaxation or noradrenaline-induced contraction in both non-pregnant and pregnant guinea pig uterine arteries. This study has shown that: (i) VIP induces relaxation of guinea pig uterine artery acting as a partial agonist on receptors localized in smooth muscle; (ii) pregnancy does not alter the response of guinea pig uterine arteries to VIP and does not change the receptor affinity for VIP, the efficiency of the receptor coupling or the VIP receptor density; and (iii) VIP does not modulate effects of neurotransmitters on guinea pig uterine arteries and pregnancy is not associated with the changes of VIP-neurotransmitter interaction.

Characterization of adenosine action in isolated rat renal artery. Possible role of adenosine A(2A) receptors.

Adenosine (0.1-300 microM) induced concentration- and endothelium-dependent relaxation of rat renal artery (RRA). N(G)-Nitro-L-arginine (L-NOARG, 10 microM) significantly reduced adenosine-elicited dilatation, but not the application of indomethacin (10 microM), ouabain (100 microM) or tetraethylammonium (TEA, 500 microM). In the presence of high concentration of K(+) (100 mM) or glibenclamide (1 microM), adenosine-evoked relaxation was almost abolished. 8-(3-Chlorostyril)caffeine (CSC, 0.3-3 microM), a selective A(2A)-antagonist, significantly reduced adenosine-evoked dilatation in a concentration-dependent manner (pA(2)=7.29). Conversely, 1,3-dipropyl-8-cyclopentylxanthine (DPCPX, 10 nM), an A(1)-antagonist, did not alter adenosine-induced relaxation. These results indicate that adenosine produces endothelium-dependent relaxation of isolated RRA. Dilatation evoked by adenosine is mediated by predominant releasing of endothelium-derived hiperpolarizing factor (EDHF) and also in one part of nitric oxide (NO) from endothelial cells. The obtained results also suggest that RRA response to adenosine is most likely initiated by activation of endothelial adenosine A(2A) receptors.

Acetylcholine-induced contractions in the porcine internal mammary artery: possible role of muscarinic receptors.

The effect of acetylcholine on the isolated, non-precontracted, porcine internal mammary artery (IMA) was investigated. Acetylcholine induced concentration-dependent contractions of non-precontracted IMA rings with denuded endothelium (pEC50 = 5.80 +/- 0.04) and was without effect on arterial segments with intact endothelium. The muscarinic receptor antagonists atropine, pirenzepine, methoctramine and p-fluoro-hexahydro-sila-diphenidol (pFHHSiD) antagonized the response to acetylcholine. The constrained pA2 values were 10.14, 7.74, 7.34 and 10.5, respectively. It is concluded that acetylcholine induces concentration-dependent contractions of porcine internal mammary artery rings on basal tone and that this contractile effect is probably due to direct cholinergic stimulation of smooth muscle cells, maybe including activation of muscarinic M1 receptors.

Characterization of oxytocin actions in guinea-pig isolated uterine artery: the effect of pregnancy.

While the contractile effect of oxytocin on uterine artery has been reported, little is known about whether pregnancy affects the responsiveness of this artery to oxytocin. If it does, is it a consequence of changed endothelial function, as has been proposed for some other vasoconstrictors. Furthermore, the receptor subtypes involved in oxytocin action on uterine artery has not been yet determined. Therefore the purposes of this study were to (1) determine the receptor subtypes involved in oxytocin action in non-pregnant and pregnant guinea-pig uterine artery and to (2) determine whether possible changes in uterine artery sensitivity to oxytocin during pregnancy are due to altered endothelial function. Therefore, the effect of oxytocin on non-pregnant and pregnant guinea-pig uterine arterial rings with and without endothelium was investigated. In non-pregnant guinea-pig uterine artery oxytocin induced contraction (pEC50 = 7.63) with greater potency than in pregnant guinea-pig uterine artery (pEC50 = 7.17). Removal of the endothelium did not affect oxytocin-induced contractions, regardless of the pregnancy status. The uterine arteries did not respond to [Thr4, Gly7]oxytocin. In the preparations studied, [d(CH2)5Tyr(Me)2]vasopressin and [d(CH2)5, D-Ile2, Ile4]vasopressin antagonized oxytocin action with the following pKB values ([d(CH2)5Tyr(Me)2]vasopressin versus [d(CH2)5, D-Ile2, Ile4]vasopressin): 8.24 versus 7.29 and 8.11 versus 7.17 for non-pregnant guinea-pig uterine artery with and without endothelium, respectively; 8.39 versus 7.25 and 8.35 versus 7.25 for pregnant guinea-pig uterine artery with and without endothelium, respectively. We suggest that, in uterine arteries, oxytocin induces contraction by activation of vasopressin V1A receptors. The potency of oxytocin in uterine artery is decreased during pregnancy and this is not associated with altered endothelial function.

Predominant role for nitric oxide in the relaxation induced by vasoactive intestinal polypeptide in human uterine artery.

It has been previously shown that vasoactive intestinal polypeptide (VIP) induces endothelium-dependent relaxation of the human uterine artery. However, the nature of the mediator of the VIP-induced endothelium-dependent relaxation of the human uterine artery has not yet been determined. Therefore these experiments were undertaken to examine the effects of VIP on human uterine arteries and to establish the role of various endothelial factors on the relaxation induced by VIP. The experiments were performed on isolated human uterine arterial rings. VIP (0.3-100 nM) induced a concentration-dependent relaxation of human uterine arteries with intact endothelium (pEC50 = 8.06+/-0.14, n = 28). After the removal of the endothelium this relaxation was abolished (n = 6). Indomethacin (10 microM), a cyclooxygenase inhibitor, and diethylcarbamazine (100 microM), a lipoxygenase blocker, had no effects on VIP-induced relaxation. In contrast, methylene blue (10 microM), a blocker of guanylate cyclase, NG-monomethyl-L-arginine (10 microM), an inhibitor of nitric oxide (NO) synthase, and 4-aminopyridine (1 mM), a non-selective blocker of K+ channels, antagonized the effect of VIP with suppression of maximal VIP-induced relaxation. Non-competitive antagonism with methylene blue revealed that the pKa value for VIP-receptor complex was 8.10+/-0.10 (n = 6) and the receptor reserve expressed as KA/EC50 was 0.89+/-0.11, where pKa = log10KA, and KA is the dissociation constant of VIP-receptor complex. Therefore, on the basis of the results presented, we can conclude that VIP induces endothelium-dependent relaxation in human uterine arteries, acting as a partial agonist on this blood vessel. It appears that endothelium-dependent relaxation induced by VIP in human uterine artery can be entirely explained by the release of NO from endothelial cells.

Effect of oxytocin as a partial agonist at vasoconstrictor vasopressin receptors on the human isolated uterine artery.

1. The effect of oxytocin on endothelium-intact and endothelium-denuded segments of the human uterine artery rings was investigated. 2. In both types of preparation oxytocin induced contraction of human uterine artery with similar potency and efficacy (pEC50 values: 6.95 +/- 0.05 vs 7.06 +/- 0.01; maximal response values: 61 +/- 4.1% vs 63 +/- 5.1% for arteries with and without endothelium, respectively). 3. In contrast, human uterine arteries, both intact and denuded of endothelium, did not respond to the addition of the selective oxytocin receptor agonist, [Thr4, Gly7]oxytocin (10 nM(-1) microM). 4. The vasopressin receptor antagonists, [d(CH2)5Tyr(Me)]AVP (10-100nM) and [d(CH2)5,D-Ile2,Ile4]AVP (300 nM-3 microM) produced parallel rightward shifts of the curves for oxytocin. The Schild plots constrained to a slope of unity gave the following -log K(B) values: [d(CH2)5Tyr(Me)] AVP vs [d(CH2)5,D-Ile2,Ile4] AVP 9.24 vs 6.91 and 9.26 vs 6.84 for human uterine artery with intact and those denuded of endothelium, respectively. In contrast, in both types of preparations the oxytocin receptor antagonist, [d(CH2)5Tyr(OMe), 2Orn8]vasotocin (1 microM), did not significantly affect oxytocin-induced contractions. 5. The calculated pK(A) values for oxytocin itself also did not differ between preparations: 6.56 and 6.43 for human uterine artery with and without endothelium, respectively. In both types of preparations, the receptor reserve (K(A)/EC50) was close to unity (intact vs denuded: 3.9 vs 3.0). 6. It is concluded that, in human uterine artery, oxytocin induces contractions that are not modulated by the endothelium. It is likely that oxytocin acts as a partial agonist on human uterine artery, regardless of the endothelial condition. On the basis of differential antagonists affinity and affinity of oxytocin itself, it is probable that receptors involved in oxytocin-induced contraction in human uterine arteries belong to the V(1A) vasopressin receptors.

Endothelium-dependent relaxation in response to acetylcholine in pregnant guinea-pig uterine artery.

Recently, strong evidence has suggested that nitric oxide (NO) synthesis is significantly increased in the uterine artery during pregnancy, which may mediate the increased blood flow to the uterus that is characteristic of pregnancy. We therefore investigated the nature of the mediators of acetylcholine (ACh)-induced relaxation in pregnant guinea-pig uterine arterial rings. ACh (0.1 nM to 60 microM) induced endothelium-dependent relaxation of phenylephrine-precontracted pregnant guinea-pig uterine artery. N(G)-monomethyl-L-arginine (3-30 microM) antagonized the effect of ACh, with suppression of maximal ACh-induced relaxation, in a concentration-dependent manner. The inhibition of relaxation by N(G)-monomethyl-L-arginine (10 microM) was significantly overcome by L-arginine (10 microM), but not by D-arginine (100 microM). On the contrary, the administration of indomethacin (10 microM) and diethylcarbamazine (100 microM) did not modify the relaxation of guinea-pig uterine artery induced by ACh. The ACh-evoked relaxation was unaltered when K+-rich Krebs-Ringer bicarbonate solution was used to induce tone instead of phenylephrine, or when a nonselective blocker of K+ channels, 4-aminopyridine (6 mM), was applied to phenylephrine-precontracted segments. It is concluded that the relaxation induced by ACh in pregnant guinea-pig uterine artery can be explained entirely by the release of NO from vascular endothelial cells, without involvement of other endothelium-derived relaxing factors, similar to that previously reported for non-pregnant guinea-pig uterine artery. Thus, it seems that increased activity of NO synthase during pregnancy is without significant influence on the ACh action on uterine artery.

Indomethacin depresses prostaglandin F2 alpha-induced contraction in guinea-pig uterine artery with both intact and denuded endoth.

The purpose of this study was to explore whether cyclooxygenase products derived from endothelium or vascular muscle participate in the response of guinea-pig uterine arterial rings to prostaglandin F2 alpha (PGF2 alpha). Contraction to PGF2 alpha (0.1-30 microM) occurred with and without endothelium at similar potency and efficacy (pEC50 (-log EC50) values respectively 5.87 +/- 0.06 and 5.97 +/- 0.07; maximal response respectively 78.1 +/- 1.3% and 76.9 +/- 1.5% of contraction induced by 126 mM KCl). Indomethacin (3-30 microM) suppressed the maximum response to PGF2 alpha and induced a rightward shift of concentration-response curves, regardless of the presence of endothelium. pIC50 values for indomethacin were 4.67 and 4.74 for vessels with and without endothelium, respectively. In contrast, the thromboxane synthesis inhibitor OKY-046 (10 and 100 microM) did not affect the response to PGF2 alpha. We conclude that the PGF2 alpha-induced contraction in guinea-pig uterine artery is mediated, at least in part, through constrictor non-thromboxane prostanoid(s) of vascular muscle origin.

Indomethacin reduces contraction of isolated non-pregnant human uterine artery induced by prostaglandin F2 alpha.

The purpose of this study was to explore whether cyclooxygenase products derived from endothelium or vascular smooth muscle participate in the response of human uterine artery to prostaglandin F2 alpha. Experiments were performed using human uterine arterial rings. Prostaglandin F2 alpha (0.4 nM-1 microM) induced contraction of human uterine arteries with both intact and denuded endothelium with similar potency and efficacy (pD2 values: 7.93 +/- 0.01 and 8.07 +/- 0.03 for vessels with and without endothelium respectively; maximal response values: 89.1 +/- 4.7% and 92.3 +/- 3.8% for vessels with and without endothelium respectively). Indomethacin (10 microM) significantly suppressed the maximum effects of prostaglandin F2 alpha and induced a shift towards the right of the prostaglandin F2 alpha concentration-response curves, regardless of the endothelial condition. On the other hand, in both types of preparations, OKY-046 (10 microM), an inhibitor of thromboxane synthesis, did not affect prostaglandin F2 alpha-induced contraction of human uterine arteries. It is concluded that in human uterine artery prostaglandin F2 alpha-induced contraction is mediated, at least in part, through constrictor prostanoid(s) of vascular smooth muscle origin that is not thromboxane A2.

Effect of the vascular endothelium on contractions induced by prostaglandin F2 alpha in isolated pregnant guinea pig uterine artery.

The purpose of this study was to examine the influence of endothelium on prostaglandin F2 alpha-mediated contractions in pregnant guinea pig uterine artery. Consequently, the effects of prostaglandin F2 alpha on pregnant guinea pig uterine arterial rings with both intact and denuded endothelium were studied. In vessels with denuded endothelium prostaglandin F2 alpha (0.1-10 microM) induced contraction (pD2 = 6.17) with greater potency than in vessels with intact endothelium (pD2 = 5.68). NG-Monomethyl-L-arginine (10 microM) did not affect the concentration-response curve for prostaglandin F2 alpha, regardless of endothelial condition. In contrast, in both types of preparation, indomethacin (10 microM) increased the maximal response value obtained with prostaglandin F2 alpha, but this effect was significantly greater in preparations with intact than in those with denuded endothelium (128.3 versus 206.5%). Moreover, indomethacin shifted the concentration-response curve for prostaglandin F2 alpha to the left only in preparations with intact endothelium. The pKA values for prostaglandin F2 alpha itself did not differ between preparations: 5.41 and 5.52 for pregnant guinea pig uterine artery with and without endothelium, respectively. The receptor reserve expressed as KA/EC50 was significantly greater in rings with denuded (4.44) compared to those in rings with intact endothelium (1.86). We conclude that prostaglandin-F2 alpha-induced contraction in pregnant guinea pig uterine artery is modulated by the vascular endothelium. It is probable that cyclooxygenase products relating to vasodilatation and derived from endothelium mediate this effect, acting as a functional endogenous antagonist and thereby reducing the apparent efficacy and potency of prostaglandin F2 alpha.

Characterization of arginine vasopressin actions in human uterine artery: lack of role of the vascular endothelium.

1. The effect of arginine vasopressin (AVP) on human uterine artery rings, both intact and denuded of endothelium, was investigated. 2. Initially, AVP (63 pM-32 nM) induced concentration-dependent contraction of human uterine artery (pD2 = 8.92 +/- 0.01). Removal of the endothelium did not affect the concentration-response curve for AVP (pD2 = 8.83 +/- 0.03). 3. In contrast, human uterine arteries, both intact and denuded of endothelium, did not respond to the addition of 1-desamino-8-D-arginine vasopressin (dDAVP, 1 nM-1 microM). 4. In both types of preparations, [d(CH2)5Tyr(Me)AVP (1-10 nM) and [d(CH2)5,D-Ile2,Ile4]AVP (300 nM-3 microM) produced parallel rightward shifts of the curves for AVP. The Schild plots constrained to a slope of unity gave the following -log KB values: [d(CH2)5Tyr(Me)]AVP vs. [d(CH2)5,D-Ile2,Ile4]AVP 9.66 vs. 6.69 and 9.61 vs. 6.80 for human uterine artery, intact and denuded of endothelium, respectively. 5. The pKA values for AVP itself also did not differ between preparations: 6.56 and 6.43 for human uterine artery with and without endothelium, respectively. In both types of preparations, the receptor reserve (KA/EC50) was considerably greater than unity (intact vs. denuded: 228 vs. 244). 6. It is concluded that, in human uterine artery, AVP induces contractions that are not modulated by the endothelium. It is likely that AVP acts as a full agonist on human uterine artery, regardless of the endothelial condition. On the basis of differential antagonists affinity and affinity of AVP itself, it is probable that vasopressin receptors involved in AVP-induced contraction in human uterine arteries belong to the V1a or V1a-like subtype.