Enhanced reactivity to vasopressin in rat basilar arteries during vasospasm after subarachnoid hemorrhage

Subarachnoid hemorrhage increases the plasma level of vasopressin, a well-known vasoconstrictor. We examined the sensitivity to vasopressin in rat basilar artery after subarachnoid hemorrhage using a rat subarachnoid hemorrhage model. Vasospasm was observed 1-2 days after subarachnoid hemorrhage induction, and the contractile response to vasopressin in rat basilar arteries was assessed. The concentration-response curve for vasopressin in subarachnoid hemorrhage (1 day) rats shifted leftward compared with that of control rats. The concentration-response curve for vasopressin V(1) receptor agonist also shifted leftward and upward compared with that of control rats. The concentration-response curve for vasopressin was inhibited not by vasopressin V(2) receptor antagonist but by vasopressin V(1) receptor antagonist. Thus, it was demonstrated that the vasoconstricting effect of vasopressin was significantly enhanced in the vasospasm phase after subarachnoid hemorrhage.     

Comparison of vasoconstrictor actions of endothelin-1 in cerebral, coronary, and mesenteric arteries of the dog.

Vasoconstrictor actions of endothelin-1 (ET) were compared between endothelium-removed strips of cerebral (basilar, posterior cerebral, and middle cerebral) and peripheral (coronary and mesenteric) arteries of the dog. ET produced a concentration-dependent contraction in these arteries. A threshold concentration and EC50 value for ET were significantly lower in the basilar, posterior cerebral, middle cerebral, and coronary arteries than in the mesenteric artery. In the basilar artery, nifedipine caused a rightward displacement of the concentration-response curve for ET with a significant reduction in the maximum response to ET. On the other hand, nifedipine showed a typical noncompetitive antagonism against ET in the mesenteric artery. Contractile responses of the mesenteric artery to ET determined under an elevation of extracellular K+ concentration were comparable to the responses of the basilar artery to this peptide determined under normal K+ concentrations. The cerebral and coronary arteries, but not the mesenteric artery, relaxed significantly from the resting level when placed in a Ca(2+)-free solution containing 0.1 mM EGTA (0-Ca solution). The readdition of Ca2+ to the cerebral and coronary arteries soaked in the 0-Ca solution caused a biphasic contraction that was susceptible to inhibition by nifedipine. When ET in concentrations below 10(-9) M was introduced before the Ca(2+)-induced contraction, this peptide produced no detectable contraction, but potentiated the Ca(2+)-induced contraction. The extent of potentiation induced by ET was much greater in the cerebral and coronary arteries than in the mesenteric artery. Even in the 0-Ca solution, higher concentrations of ET (1 x 10(-8) and 3 x 10(-8) M) produced a contraction that was weaker in the basilar artery than in the mesenteric artery. These results indicate that the cerebral and coronary arteries exhibited more potent contractions in response to lower concentrations (below 10(-9) M) of ET than the mesenteric artery. A likely possibility for these enhanced responses to ET in the cerebral and coronary arteries appears to be that the voltage-dependent Ca2+ channels in these arteries are more activated in the resting state than those in the mesenteric artery.     

Ca(2+) uptake function of sarcoplasmic reticulum during contraction of rat arterial smooth muscles

To determine the Ca(2+) uptake function of the sarcoplasmic reticulum during contraction, the effects of cyclopiazonic acid or thapsigargin, agents that inhibit sarcoplasmic reticulum Ca(2+)-ATPase, on the contractile responses to K(+) or norepinephrine were compared in endothelium-denuded strips of femoral, mesenteric and carotid arteries of the rat. The addition of K(+) (3-20 mM) to the strips caused a concentration-dependent contraction, and the sensitivity to K(+) was much higher in the carotid artery than in the other arteries. The preincubation of strips with cyclopiazonic acid (10 microM) or thapsigargin (100 nM) caused a leftward shift of the concentration-response curve for K(+), and this effect was smaller in the carotid artery than in the other arteries. Inhibition of sarcoplasmic reticulum Ca(2+) uptake caused the sensitivity to K(+) to be similar in the three arteries. Similar results were obtained when the contractile responses to norepinephrine were determined. Cyclopiazonic acid itself induced similar transient contractions in the three arteries. The addition of caffeine (20 mM) caused a transient contraction that was smaller in the carotid artery than in the other arteries. We conclude that (1) the Ca(2+) influx during stimulation with K(+) or norepinephrine is buffered by the sarcoplasmic reticulum in femoral and mesenteric arteries, (2) this function is weak in the carotid artery, probably because the sarcoplasmic reticulum of this artery is almost filled with Ca(2+) in the resting state, and (3) the Ca(2+) uptake function of the sarcoplasmic reticulum during contraction is reflected by the contractile sensitivity in these arteries.     

Vascular effects of carvedilol, a new beta-adrenoceptor antagonist with vasodilating properties, in isolated canine coronary artery

The pharmacologic properties of carvedilol, a beta-adrenoceptor antagonist with vasodilating activity, were investigated in isolated canine coronary artery ring preparations. Carvedilol competitively antagonized the relaxations caused by isoproterenol in 40 mM K+-depolarized preparations and its pA, value was 9.70 +/- 0.08. At concentrations greater than or equal to 3 x 10(-6) M, carvedilol significantly inhibited the contractile response to high [K+]o. Compared with the inhibitory effect on the KCl-induced contraction, the drug was less effective in suppressing the contraction induced by prostaglandin F2 alpha (PGF2 alpha). In addition, carvedilol (10(-7) to 3 x 10(-5) M) suppressed the contraction produced by Bay K 8644, a Ca2+ channel agonist. The concentration-response curve for Bay K 8644 was shifted downward by carvedilol in a concentration-dependent manner. The drug also produced a concentration-dependent inhibitory effect on the 4-aminopyridine-induced rhythmic contractions in a similar fashion to the Ca2+ channel antagonists. Carvedilol was ineffective in suppressing the contractions induced by PGF2 alpha in Ca2+-free solution and by A-23187. The results suggest that carvedilol exerts a vasodilating action possibly by inhibiting Ca2+ influx through potential-operated Ca2+ channels, although the concentrations required for producing the vasodilation are much higher than that for the beta-adrenoceptor antagonism in canine coronary artery.     

alpha,beta-MeATP augments the UTP contraction of rabbit basilar artery

The mechanism underlying the interaction between alpha,beta-methyleneadenosine 5'-triphosphate (alpha,beta-MeATP) and uridine 5'-triphosphate (UTP) was investigated using the basilar artery of a rabbit. UTP induced a concentration-dependent contraction, whereas P2X receptor agonists, such as alpha,beta-MeATP and 2-methylthioadenosine 5'-triphosphate (2-MeSATP), did not induce any contraction up to 100 microM. alpha,beta-MeATP augmented the UTP contraction two-fold, immediately and reversibly. This effect was observed with ectonucleotidase inhibition with 1 mM Ni(2+), the removal of extracellular Ca(2+) or Evans blue. The contractile response to adenosine 5'-O-(3-triphosphate) (ATPgammaS), a selective agonist for P2Y(4), was augmented by pretreatment with alpha,beta-MeATP also. ATPgammaS had no additional effect on the UTP contraction fully activated with alpha,beta-MeATP. UTP (100 microM) did not induce an increase in cytosolic Ca(2+) in a rabbit basilar arterial strip; however, in the presence of 1 mM alpha,beta-MeATP, UTP induced a significant increase in cytosolic Ca(2+). These results suggest that alpha,beta-MeATP facilitates the activation by UTP of the P2Y receptor (P2Y(4)) of the rabbit basilar artery through mechanisms other than nucleotidase inhibition, and that it does not do so via a P2X receptor.     

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.     

Regional differences in the arterial response to vasopressin: role of endothelial nitric oxide.

1. The isometric response to arginine-vasopressin (10(-10)-10(-7)M) was studied in 2 mm long rabbit arterial segments isolated from several vascular beds (cutaneous, pial, renal, coronary, muscular, mesenteric and pulmonary). 2. Vasopressin induced contraction in central ear (cutaneous), basilar (pial), renal, coronary and saphenous (muscular) arteries, but had no effect in mesenteric and pulmonary arteries; the order of potency for the contraction was: ear > basilar > renal > coronary > saphenous arteries. 3. Treatment with the blocker of nitric oxide synthesis NG-nitro-L-arginine methyl ester (L-NAME; 10(-6)-10(-4) M) increased significantly (P < 0.05) the contraction to vasopressin in ear (148% of control), basilar (150% of control), renal (304% of control), coronary (437% of control) and saphenous (235% of control) arteries. Removal of the endothelium increased significantly (P < 0.05) the contraction to vasopressin in basilar (138% of control), renal (253% of control), coronary (637% of control) and saphenous (662% of control) arteries, but not in ear artery. Mesenteric and pulmonary arteries in the presence of L-NAME or after endothelium removal did not respond to vasopressin, as occurred in control conditions. 4. The specific antagonist for V1 vasopressin receptors d(CH2)5Tyr(Me)AVP (3 x 10(-9)-10(-7) M) was more potent (pA2 = 9.3-10.1) than the antagonist for both V1 and V2 vasopressin receptors desGly-d(CH2)5-D-Tyr(Et)ValAVP (10(-7)-10(-6) M) (pA2 = 7.4-8.4) to block the contraction to vasopressin of ear, basilar, renal and coronary arteries. 5. The specific V2 vasopressin agonist [deamino-Cys1, D-Arg8]-vasopressin (desmopressin) (10(-10)-10(-7) M) did not produce any effect in any effect in any of the arteries studied, with or without endothelium. 6. In arteries precontracted with endothelin-1, vasopressin or desmopressin did not produce relaxation. 7. These results suggest: (a) most arterial beds studied (5 of 7) exhibit contraction to vasopressin with different intensity; (b) the vasoconstriction to this peptide is mediated mainly by stimulation of V1 vasopressin receptors, and (c) endothelial nitric oxide may inhibit the vasoconstriction to this peptide, especially in coronary and renal vasculatures.     

Cysteinyl leukotrienes and leukotriene B mediate vasoconstriction to arginine vasopressin in rat basilar artery

Arginine vasopressin (AVP) has been reported to be involved in the development of cerebral vasospasm after haemorrhage and cerebral oedema following ischaemia. Endogenously produced 5-lipoxygenase metabolites are able to contract isolated endothelium-preserved arterial strips and modulate vascular permeability. The present study addresses the role of 5-lipoxygenase and its products, namely cysteinyl leukotrienes (CysLTs) and leukotriene (LT) B4, in the contraction induced by AVP in rat basilar artery. Contractile responses to LTD4, LTC4, LTB4 or AVP were assessed in spiral preparations of rat endothelium-intact basilar artery. Contractions to AVP were determined in the absence or presence of 5-lipoxygenase inhibitors or CysLT1 or BLT receptor antagonists. Contractile responses to leukotrienes and AVP are expressed as a percentage of the contraction induced by 80 mmol/L KCl. Leukotriene D4, LTC4 and LTB4 acted as vasoconstrictor agents in rat basilar artery, causing contractions (all at concentrations of 1 micromol/L) of 42 +/- 13, 54 +/- 15 and 25 +/- 6% of the response to 80 mmol/L KCl, respectively. A concentration-response curve was constructed for AVP over the range 1 pmol/L to 10 nmol/L and an EC50 value of 0.19 +/- 0.02 nmol/L (n = 30) was determined. The presence of the 5-lipoxygenase inhibitors ZM 230487 (10 nmol/L and 0.1 and 1 micromol/L) and AA 861 (1, 3, 10, and 30 micromol/L), the CysLT1 receptor antagonist MK 571 (3, 10 and 30 micromol/L) or the BLT receptor antagonists CP 105696 and LY 255283 (3, 10 and 30 micromol/L for both) in the organ bath significantly attenuated the contractions induced by AVP in rat basilar artery (P < 0.05). The experimental results of the present study provide the first evidence for the involvement of CysLTs and LTB4 in the in vitro constriction induced by AVP in rat basilar artery. In the context of previously reported involvement of AVP in the development of cerebral vasospasm and oedema, the present study draws attention to the potential role played by the 5-lipoxygenase pathway in these pathological processes.     

Relative contribution of intracellular and extracellular Ca2+ to alpha2-adrenoceptor-mediated contractions of ovine pulmonary artery.

We have examined the mechanism of contractions elicited by guanfacine, a selective agonist for alpha(2A/D)-adrenoceptors and its modulations by cyclic nucleotides in isolated ovine resistance intra-pulmonary artery. Guanfacine (10 nM-30 microM) produced concentration-dependent contraction of the pulmonary artery rings mounted for isometric recording. Yohimbine (0.1 microM), a nonspecific alpha(2)-adrenoceptor antagonist caused a parallel shift to the right (1.2 log unit) in the concentration-response curve of guanfacine without depressing the maxima. Preincubation of the tissues with Ca(2+)-free solution (EGTA 1mM) for 30 min caused a rightward shift (0.8 log unit) of the concentration-response curve of guanfacine with the inhibition of the maxima by 30+/-4.6%. L-type calcium channel blocker, nifedipine (1 microM) slightly inhibited (20%) the maximal contraction elicited with guanfacine (10 microM). On the other hand, brief exposure to cyclopiazonic acid (10 microM), an inhibitor of IP3-sensitive sarcoplasmic reticulum Ca(2+)-ATPase, resulted in marked inhibition of concentration-dependent contractions elicited with guanfacine (10 nM-30 microM), with the maxima being inhibited by 51+/-3.11%. In addition, agents that increase intracellular cAMP and cGMP suppressed guanfacine-induced contractions. The results of the present study suggest that alpha(2)-adrenoceptor-mediated contractions in ovine resistance pulmonary artery is primarily dependent on intracellular Ca(2+) with a small contribution from Ca(2+)-influx through voltage-dependent L-type calcium channels.     

Organic Ca(2+)-antagonist-resistant response to FMRF-NH2 on the molluscan smooth muscle.

1. FMRF-amide (10(-7)-10(-5) M) contracted molluscan anterior byssus retractor muscle in a concentration-dependent fashion. 2. The concentration-response curve of FMRF-amide was shifted rightward by an analogue of FMRF-amide, FMRf-amide ([D-Phe4]FMRF-amide, putative FMRF-amide receptor antagonist) in a parallel manner (pA2 = 4.87 +/- 0.04). 3. Although a contractile response to KCl was reduced by the organic Ca2+ antagonists (verapamil, diltiazem and high concentration of nifedipine and nicardipine). FMRF-amide-induced contraction was not markedly reduced by them. 4. In the Ca(2+)-free medium, FMRF-amide-induced contraction was diminished. The response was also reduced by TMB-8 (10(-4) M), suggesting that FMRF-amide-induced contraction might be partly dependent on intracellular Ca2+. 5. An inorganic Ca(2+)-antagonist, MnCl2, markedly reduced the FMRF-amide- and KCl-induced contraction. The results show that FMRF-amide-induced contraction might be dependent on extracellular Ca2+. 6. These findings suggest that FMRF-amide-induced contraction might be mediated through an action on FMRF-amide receptors and not through the activation of organic Ca2+ antagonist-sensitive Ca2+ channels.     

Different calcium-mobilizing receptors share the same guanine nucleotide-binding protein pool in hepatocytes.

High affinity binding of epinephrine to the alpha 1-adrenoceptor reflects the association of the ligand-receptor complex with a guanine nucleotide-binding protein (G protein) and thereby allows the receptor-G protein interaction to be assessed by radioligand binding methods. We have used [3H]prazosin/epinephrine competition binding to rat liver plasma membranes to examine the effects of other Ca(2+)-mobilizing hormones on the interaction between the alpha 1-adrenoceptor and its G protein. The aim of our experiments was to test whether the different Ca(2+)-mobilizing receptors in liver share the same limited pool of G proteins. [Arg8] Vasopressin (AVP) caused a concentration-dependent (EC50 = 0.49 +/- 0.03 nM) inhibition of the extent to which epinephrine formed a high affinity complex with the alpha 1-adrenoceptor; antagonist binding was unaffected by AVP. The effect of AVP was competitively antagonized (Kd = 0.27 +/- 0.10 nM) by a selective peptide antagonist of the V1 vasopressin receptor. We conclude that, in rat hepatocytes, alpha 1-adrenoceptors and V1 vasopressin receptors converge to interact with the same pool of G proteins.     

Characterization of a novel nonpeptide vasopressin V(2)-agonist, OPC-51803, in cells transfected human vasopressin receptor subtypes

We discovered the first nonpeptide arginine-vasopressin (AVP) V(2)-receptor agonist, OPC-51803. Pharmacological properties of OPC-51803 were elucidated using HeLa cells expressing human AVP receptor subtypes (V(2), V(1a) and V(1b)) and compared with those of 1-desamino-8-D-arginine vasopressin (dDAVP), a peptide V(2)-receptor agonist. OPC-51803 and dDAVP displaced [(3)H]-AVP binding to human V(2)- and V(1a)-receptors with K(i) values of 91.9+/-10.8 nM (n = 6) and 3.12+/-0.38 nM (n = 6) for V(2)-receptors, and 819+/-39 nM (n = 6) and 41.5+/-9.9 nM (n = 6) for V(1a)-receptors, indicating that OPC-51803 was about nine times more selective for V(2)-receptors, similar to the selectivity of dDAVP. OPC-51803 scarcely displaced [(3)H]-AVP binding to human V(1b)-receptors even at 10(-4) M, while dDAVP showed potent affinity to human V(1b)-receptors with the K(i) value of 13.7+/-3.2 nM (n = 4). OPC-51803 concentration-dependently increased cyclic adenosine 3', 5'-monophosphate (cyclic AMP) production in HeLa cells expressing human V(2)-receptors with an EC(50) value of 189+/-14 nM (n = 6). The concentration-response curve for cyclic AMP production induced by OPC-51803 was shifted to the right in the presence of a V(2)-antagonist, OPC-31260. At 10(-5) M, OPC-51803 did not increase the intracellular Ca(2+) concentration ([Ca(2+)](i)) in HeLa cells expressing human V(1a)-receptors. On the other hand, dDAVP increased [Ca(2+)](i) in HeLa cells expressing human V(1a)- and V(1b)-receptors in a concentration-dependent fashion. From these results, OPC-51803 has been confirmed to be the first nonpeptide agonist for human AVP V(2)-receptors without agonistic activities for V(1a)- and V(1b)-receptors. OPC-51803 may be useful for the treatment of AVP-deficient pathophysiological states and as a tool for AVP researches.     

Effect of a new antiarrhythmic BK 129 on canine vascular smooth muscle in vitro.

The effect of a new carbanilate derivative BK 129, 1-methoxymethyl-2(1-perhydroazepinyl)ethyl ester 2-(n)-pentyloxycarbanilic acid hydrochloride, was tested on preparations of canine dorsal pedal and coronary artery in vitro. The drug produced relaxation of arterial rings both under resting conditions and under precontraction with KCl. BK 129, at a concentration of 10(-5) mol/l, produced a shift to the right of the concentration-response curve of phenylephrine, with a decrease in the slope and a depression of the maximal response. BK 129 inhibited the response to norepinephrine and blocked histamine- and norepinephrine-induced contractions in Ca(2+)-free physiological salt solution. The electrical stimulation-induced contractions of arterial preparation were also inhibited. BK 129 shifted the concentration-response curve of CaCl2 in Ca(2+)-free depolarizing solution to the right in a non-competitive manner. It may be concluded that BK 129 is a local anesthetic which possesses relaxant properties. It appears to inhibit Ca2+ entry into the smooth muscle cell and Ca2+ release from sarcoplasmic reticulum.     

Hypertrophic growth of cultured neonatal rat heart cells mediated by vasopressin V(1A) receptor

Primary cultures of neonatal cardiac myocytes were used to determine both the identity of second messengers that are involved in vasopressin receptor-mediated effects on cardiac hypertrophy and the type of vasopressin receptor that is involved in vasopressin-induced cell growth. Neonatal rat myocytes were plated at a density of 1x10(6) cells per 60 mm dish and were incubated with serum-free medium for 7 days. Treatment of myocytes with vasopressin significantly increased the RNA-to-DNA ratio, by 18-25%, at culture days 4-6 and the protein-to-DNA ratio by 18-20% at culture days 5-7. Rates of protein synthesis were determined to assess their contribution to protein contents during myocyte growth. Vasopressin significantly accelerated rates of protein synthesis by 25% at culture day 6. Intracellular free Ca(2+) ([Ca(2+)](i)) was transiently increased after vasopressin exposure. After the peak increase in [Ca(2+)](i) at less than 30 s, there was a sustained increase for at least 5 min. The specific activity of protein kinase C in the particulate fraction was increased rapidly after exposure to vasopressin, and its activity remained higher for 30 min, returning to its control level within 60 min. The activity of protein kinase C in the cytosol was significantly decreased at all times after exposure to vasopressin. After vasopressin treatment, the content of c-fos mRNA was increased. The stimulatory effects of vasopressin on these parameters were significantly inhibited by vasopressin V(1A) receptor antagonist, OPC-21268, but not by vasopressin V(2) receptor antagonist, OPC-31260. These results suggest that vasopressin directly induces myocyte hypertrophic growth via the V(1A) receptor in neonatal rat heart cells.     

Effects of the calcium channel facilitator, CGP 28,392, on different modes of contraction in smooth muscle of rabbit and rat aortae and guinea-pig taenia caeci.

Effects of a Ca2+ channel facilitator, CGP 28,392, on smooth muscle contractions were examined in order to delineate characteristics of Ca2+ channels in rabbit and rat aortae and guinea-pig taenia caeci. Application of increasing concentrations of KCl induced contractile responses in these smooth muscles and CGP 28,392 shifted the concentration-response curve for KCl to the left. The maximum response was also increased in rat aorta and guinea-pig taenia. CGP 28,392 also shifted the concentration-response curves for noradrenaline in rat aorta and for histamine in taenia to the left and increased the maximum response in rat aorta. However, the corresponding curve for noradrenaline in rabbit aorta was not affected by CGP 28,392. The sustained contractions induced by KCl were inhibited by cumulative application of verapamil in these smooth muscles. Pretreatment of the muscle with CGP 28,392 decreased the inhibitory effect of verapamil. The noradrenaline-induced contraction in rat aorta and the histamine-induced contraction in taenia were also inhibited by verapamil, and CGP 28,392 antagonized the effect of verapamil. The noradrenaline-induced contraction in rabbit aorta was only slightly inhibited by verapamil, and CGP 28,392 did not modify the effect of verapamil. In these smooth muscles, cumulative application of Ca2+ to the Ca2+-depleted, KCl-treated muscle induced contraction, and the concentration-response curve for Ca2+ was shifted to the left by CGP 28,392 and to the right by verapamil. The concentration-response curves for Ca2+ in Ca2+-depleted, noradrenaline-treated rabbit and rat aortae and in Ca2+-depleted, histamine-treated taenia were also shifted to the left by CGP 28,392 and to the right by verapamil.(ABSTRACT TRUNCATED AT 250 WORDS)     

Vasopressin receptor subtypes on mesenteric and cremasteric arterioles in rat

We studied the effects of a selective vasopressin V(1A) receptor antagonist [1-(1-(4-(3-acetylaminopropoxy)benzoyl)-4-piperidyl)-3, 4-dihydro-2(1H)-quinolinone (OPC-21268)] and a selective vasopressin V(2) receptor antagonist [5-dimethylamino-1(4-(2-methylbenzoylamino)benzoyl)-2,3,4, 5-tetrahydro-1H-benzazepine (OPC-31260)] on vasopressin-induced contraction of mesenteric and cremasteric arterioles in urethane-anaesthetized rats. Vasopressin was infused intravenously for 60 min or applied topically to arterioles directly. Vasopressin infusion (50, 100 or 500 ng/kg/min) decreased the diameter of both mesenteric and cremasteric arterioles. Vasopressin (500 ng/kg/min)-induced vasoconstriction was antagonized by OPC-21268 (0. 2, 1.0 and 5.0 mg/kg, i.v.), dose-dependently, but not by OPC-31260. Topically applied vasopressin (4.6x10(-10)-4.6x10(-8) M) dose-dependently constricted both microvessels. Pre-administration of OPC-21268 (5.0 mg/kg, i.v.) completely inhibited topically applied vasopressin-induced vasoconstriction in both microvessels, and OPC-31260 partially inhibited it in cremasteric arterioles. These results suggest that vasopressin induces vasoconstriction in rat mesenteric and cremasteric arterioles mainly by stimulating vasopressin V(1A) receptors, while vasoconstriction in cremasteric arterioles is partly associated with stimulation of vasopressin V(2) receptors.     

吡那地尔,cromakalim和NS1619对电刺激所致大鼠输精管收缩的作用

目的：本文研究新近研制的ＡＴＰ敏感性钾通道开放剂，吡那地尔（Ｐｉｎ）和ｃｒｏｍａｋａｌｉｎ（Ｃｒｏ），以及钙离子激活性钾通道开放剂ＮＳ１６１９对电场刺激所致大鼠输业管收缩的作用，方法：利用电场刺激（０．３Ｈｚ，１ｍｓ，６０Ｖ）反复性引致输精管单相性收缩，结果：Ｐｉｎ和Ｃｒｏ浓度依赖性减低电刺激收缩，格列本脲（Ｇｌｉ）而非ｃｈａｒｙｂｄｏｔｏｘｉｎ拮抗上述两药的舒张的作用，Ｐｉｎ右移去甲肾上腺素的浓     

Effects of YM218, a nonpeptide vasopressin V(1A) receptor-selective antagonist, on vasopressin-induced growth responses in human mesangial cells

Mesangial cells are centrally-located glomerular pericytes with contractile, endocrine, and immunity-regulating functions. These cells are thought to maintain normal glomerular function, since mesangial cell proliferation and extracellular matrix formation are hallmarks of chronic glomerular disease. Vasopressin causes mesangial cell contraction, proliferation and hypertrophy. Consequently, the effects of YM218, a potent, nonpeptide vasopressin V(1A) receptor-selective antagonist, on the growth responses of human mesangial cells to vasopressin were investigated. YM218 showed high affinity for vasopressin V(1A) receptors, exhibiting a K(i) value of 0.18 nM. Vasopressin concentration-dependently increased intracellular Ca(2+) levels and induced hyperplasia and hypertrophy in cultured mesangial cells, YM218 potently inhibited these vasopressin-induced responses. These results clearly show that YM218 has both strong affinity for human mesangial cell vasopressin V(1A) receptors and great potency in inhibiting the vasopressin-induced growth responses of mesangial cells controlled by the vasopressin V(1A) receptors. The hyperplasia and hypertrophy of mesangial cells in vitro caused by vasopressin indicate its possible in vivo role in glomerular disease pathogenesis. Therefore, YM218 is a potent pharmacologic probe to investigate the physiologic and pathophysiologic roles of vasopressin in the development of renal disease.     

Resistance to antagonism of atrial P(1) purinoceptor responses in the presence of K(+) channel blockade

The rate of onset of the negative inotropic responses of guinea-pig isolated paced atria to the adenosine receptor agonist, N(6)-cyclopentyladenosine, was significantly slowed by the K(+) channel inhibitor, 4-aminopyridine (10 mM). The concentration-dependent inhibition of developed tension by N(6)-cyclopentyladenosine, however, was unaffected by 4-aminopyridine (10 mM). Thus, K(+) efflux only governs the speed of onset of the negative inotropic response and does not appear to be a major component in the negative inotropy produced by the adenosine A(1) receptor agonist. The P(1) purinoceptor antagonist, 8(p-sulfophenyl) theophylline (1 x 10(-5) M) significantly shifted the concentration-response curve for N(6)-cyclopentyladenosine to the right (concentration-ratio, 7.1+/-1.5). In the presence of 4-aminopyridine (10 mM), 8(p-sulfophenyl) theophylline caused a non-parallel rightwards shift of the curve. At the IC(35) there was no significant shift, whereas at the IC(75) there was a small significant displacement of the curve. The adenosine A(1)/A(3) receptor agonist, N(6)-2-(4-aminophenyl) ethyladenosine (APNEA) yielded a biphasic concentration-response curve which was significantly shifted to the right by 8(p-sulfophenyl) theophylline (1 x 10(-5 ) M). In the presence 4-aminopyridine, however, there was no shift of the APNEA concentration-response curve by 8(p-sulfophenyl)theophylline. These results show that when K(+) channels are blocked by 4-aminopyridine, the residual response is resistant to antagonism by the P(1) purinoceptor antagonist, 8(p-sulfophenyl)theophylline. This residual component may involve L-type Ca(2+) channels, the adenosine A(1) receptor being possibly coupled to the two transduction pathways for negative inotropism via the different components of the G protein (receptor-transducer promiscuity).