Biomedical subjects

J R Vane

Publications and source records attributed to J R Vane.

At least 73 records · Page 4Linked to original sources

PubMed1995

Cytokine regulation of endothelin-1 release from bovine aortic endothelial cells.

The action of cytokines on the endothelium is a pivotal event in a number of vascular pathologies and inflammatory conditions. Here we have tested the effect of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta, and IL-2 on endothelial-1 (ET-1) release from cultured bovine aortic endothelial cells. TNF-alpha and IL-1 beta caused concentration-dependent increases in ET-1 release. IL-2 up to concentrations of 100 ng/ml did not stimulate ET-1 release. The induction of ET-1 synthesis by cytokines may be of underlying importance in the endothelial cell dysfunction observed in a number of vascular diseases.

Animals↗

PubMed1994

The endothelin ETB receptor agonist, IRL 1620, causes vasodilatation and inhibits ex vivo platelet aggregation in the anaesthetised rabbit.

The ETB receptor agonist, IRL 1620 injected into the left ventricle, induced a biphasic (transient fall followed by a secondary fall lasting 5 min) depressor response in the anaesthetised rabbit. This was followed by a rise in systemic blood pressure which returned to control levels within 30 min. In addition, IRL 1620 caused a strong, transient inhibition of ADP-induced platelet aggregation ex vivo. The selective ETA receptor antagonist, FR 139317 had no effect on either the haemodynamic or anti-aggregatory effects of IRL 1620. Indomethacin which had no effect on the initial transient depressor response induced by IRL 1620, abolished the secondary fall revealing a sustained pressor response. Indomethacin also prevented the IRL 1620-induced inhibition of platelet aggregation ex vivo. Thus, IRL 1620 is a selective ETB receptor agonist in the anaesthetised rabbit, causing vasodilatation, vasoconstriction and inhibition of ex vivo platelet aggregation. The inhibition of platelet aggregation by IRL 1620 is due to an ETB receptor-mediated release of prostacyclin into the circulation, for it is inhibited by indomethacin, but not by FR 139317.

Anesthesia↗

PubMed1994

Modification of metabolism of transplantable and spontaneous murine tumors by the nitric oxide synthase inhibitor, nitro-L-arginine.

PURPOSE: To determine the effects of the nitric oxide synthase inhibitor, nitro-L-arginine on energy metabolism in transplantable and spontaneous murine tumors. METHODS AND MATERIALS: The responses of the transplantable murine tumor SCCVII/Ha and a range of spontaneously arising murine mammary adenocarcinomas to 10 mg/kg IV nitro-L-arginine were examined using in vivo 31P magnetic resonance spectroscopy (MRS). The influence of Hypnorm/Hypnovel anesthesia on the response to nitro-L-arginine was also determined in the SCCVII/Ha tumors. Data were expressed as changes in the inorganic phosphate peak area relative to the sum of all peak areas from the 31P MR spectrum, or Pi/total. RESULTS: Nitro-L-arginine at 10 mg/kg IV increased Pi/total 2-3-fold in the SCCVII/Ha tumors for at least 2 h after administration, in both anesthetized and nonanesthetized mice, consistent with increased tumor hypoxia. Similar increases in Pi/total were observed after 10 mg/kg IV nitro-L-arginine in 13 spontaneous murine tumors from three different mouse strains, where anesthetic was used. CONCLUSION: The results indicate that tumor metabolism may be modified by an inhibitor of nitric oxide synthesis, that this modification occurs in both transplantable and spontaneous murine tumors and is not affected by anesthetic.

Amino Acid Oxidoreductases↗

PubMed1994

Activation of soluble guanylyl cyclase by a factor other than nitric oxide or carbon monoxide contributes to the vascular hyporeactivity to vasoconstrictor agents in the aorta of rats treated with endotoxin.

We have examined the role of soluble guanylyl cyclase and possible mediators of its activation in the vascular hyporeactivity caused by bacterial endotoxin (lipopolysaccharide, LPS) ex vivo. Treatment of rats with E. coli LPS (10 mg/kg, i.v. for 3h) resulted in a significant reduction in the contractions elicited by norepinephrine (NE; 10(-9)-10(-6) M) in endothelium-denuded aortic rings ex vivo. Methylene blue or LY-83583, inhibitors of soluble guanylyl cyclase, completely restored contractions to NE, whereas the nitric oxide synthase (NOS) inhibitor, N omega-nitro-L-arginine methyl ester (L-NAME), caused only a partial restoration. Zinc protoporphyrin-IX, an inhibitor of heme oxygenase, did not enhance NE-induced contraction in rings from LPS-treated rats, indicating that the production of carbon monoxide (CO) does not contribute to this vascular hyporeactivity. Indomethacin, an inhibitor of cyclooxygenase, further suppressed the contractions in rings from LPS-treated rats. These results suggest that hyporesponsiveness to NE caused by LPS is due to the activation of soluble guanylyl cyclase, which is partially mediated by N(O), but not by CO. Moreover, LPS may induce the production of another mediator(s) that activate soluble guanylyl cyclase in the vascular smooth muscle.

Animals↗

PubMed1994

Antagonism of renal and systemic responses to endothelin-1 infusion with PD 145065.

The antagonism by PD 145065 (Ac-D-Bhg-L-Leu-L-Asp-L-Ile-L-Ile-L-Trp; D-Bhg = 5H-dibenzyl[a,d]cycloheptene-10,11-dihydroglycine), a non-selective endothelin ETA and ETB receptor antagonist, of the pressor and renal haemodynamic responses to an infusion of endothelin-1 was investigated in the anaesthetised rat. Infusion of endothelin-1 at 2 or 4 pmol/min/rat for 2 h produced a significant increase in mean arterial blood pressure and renal vascular resistance. Glomerular filtration rate was reduced by both infusion rates of endothelin-1 (-34% for the lower rate and -72% for the higher rate), but only the high rate significantly reduced renal blood flow (-64%). These effects of endothelin-1 were completely blocked by infusion of PD 145065 (0.05 mg/min/rat), demonstrating the efficacy of this antagonist in preventing the actions of endothelin-1 in vivo. These results lend support to an involvement of endothelin ETB receptors in the renal effects of endothelin-1.

Amino Acid Sequence↗

PubMed1994

Endothelin-1 release from endothelial cells in culture is elevated both acutely and chronically by short periods of mechanical stretch.

The effects were examined of mechanical stretch on the release of endothelin-1 (ET-1) and prostacyclin (measured as 6-keto-prostaglandin (PG) F1 alpha) from cultured endothelial cells. Stretching (0.2 Hz) for 20, 60 or 360 min caused immediate (< or = 20 min) and secondary (up to 360 min) increases in ET-1 release. The secondary but not immediate release of ET-1 was prevented by actinomycin D (8 x 10(-7) M) or cycloheximide (3.6 x 10(-6) M). Neither compound affected the release of ET-1 from unstretched cells over 360 min. Stretching of the endothelial cells increased the accumulation of 6-keto-PGF1 alpha at 360 min but not at 20 min, suggesting that stretch does not produce a rapid, non-selective increase in autacoid production from endothelial cells. The intracellular amounts of ET-1 were approximately 20 times greater than those of big ET-1. Thus, endothelial cells contain stores of ET-1 that are released rapidly by stretch.

6-Ketoprostaglandin F1 alpha↗

PubMed1994

Inducible isoforms of cyclooxygenase and nitric-oxide synthase in inflammation.

Cyclooxygenase (COX) converts arachidonic acid to prostaglandin H2, which is further metabolized to prostanoids. Two isoforms of COX exist: a constitutive (COX-1) and an inducible (COX-2) enzyme. Nitric oxide is derived from L-arginine by isoforms of nitric-oxide synthase (NOS; EC 1.14.13.39): constitutive (cNOS; calcium-dependent) and inducible (iNOS; calcium-independent). Here we have investigated inducible isoforms of COX and NOS in the acute, chronic, and resolving stages of a murine air pouch model of granulomatous inflammation. COX and NOS activities were measured in skin samples in the acute phase, up to 24 h. Activities in granulomatous tissue were measured at 3, 5, 7, 14, and 21 days for the chronic and resolving stages of inflammation. COX-1 and COX-2 proteins were assessed by Western blot. COX activity in the skin increased over the first 24 h and continued to rise up to day 14. COX-2 protein rose progressively, also peaking at day 14. COX-1 protein remained unaltered throughout. The iNOS activity increased over the first 24 h in the skin, with a further major increase in the granulomatous tissue between days 3 and 7, followed by a decrease at day 14 and a further increase at day 21. The rise in COX and NOS activities in the skin during the acute phase reinforces the proinflammatory role for prostanoids and suggests one also for nitric oxide. However, in the chronic and resolving stages, a dissociation of COX and NOS activity occurred. Thus, there may be differential regulation of these enzymes, perhaps due to the changing pattern of cytokines during the inflammatory response.

Amino Acid Oxidoreductases↗

PubMed1994

Comparison of PD 145065 and Ro 46-2005 as antagonists of contractions of guinea-pig airways induced by endothelin-1 or IRL 1620.

Cumulative concentrations of endothelin-1 or IRL 1620 (Suc-[Glu9,Ala11,15]endothelin-1-(8-21)), and endothelin ETB receptor selective agonist were similarly potent in contracting guinea-pig upper bronchi, whereas endothelin-1 was more potent than IRL 1620 in contracting lung parenchymal strips. PD 145065 (10(-5) M and 10(-4) M), a non-selective endothelin ETA/ETB receptor antagonist (Ac-D-Bhg-L-Leu-L-Asp-L-Ile-L-Ile-L-Trp), significantly attenuated the contractions induced by endothelin-1 in either preparation whereas Ro 46-2005 (3 x 10(-5) M and 10(-4) M), a non-peptide non-selective receptor antagonist (4-tert-butyl-N-[6-(2-hydroxy-ethoxy)-5-3-methoxy-phenoxy)-4-pyrimidinyl (benzenesulfonamide)), was without effect. PD 145065 (10(-6) M and 10(-5) M) also strongly inhibited contractions induced by IRL 1620, whereas Ro 46-2005 caused much less antagonism. Thus, PD 145065 is a more potent antagonist than Ro 46-2005 of contractions induced by endothelin-1 or IRL 1620 and mediated by endothelin ETB receptors in these guinea-pig airway preparations.

Animals↗

PubMed1994

The Croonian Lecture, 1993. The endothelium: maestro of the blood circulation.

The vascular endothelium plays a vital role in the control of the circulation. It metabolizes various vasoactive substances, coverts angiotensin I to angiotensin II and secretes the potent vasodilators prostacyclin and EDRF (NO) and the vasoconstrictor peptide endothelin-1. The balance between these mediators determines the responses of the cardiovascular system in diseases such as hypertension, atherosclerosis and myocardial infarction.

Animals↗

PubMed1994

Attenuation of the induction of nitric oxide synthase by endogenous glucocorticoids accounts for endotoxin tolerance in vivo.

An enhanced formation of nitric oxide (NO) due to induction of a calcium-independent (inducible) NO synthase (iNOS) contributes importantly to the cardiovascular failure caused by bacterial endotoxin. Repeated challenges with small doses of endotoxin result in tolerance to both peripheral vascular failure and death caused by subsequent injection of a higher dose of endotoxin. Here we investigate whether tolerance to endotoxin is associated with a lack of induction of iNOS in vivo and whether endogenous glucocorticoids play a role in the development of endotoxin tolerance. In anesthetized rats, i.v. administration of Escherichia coli endotoxin [lipopolysaccharide (LPS); 2 mg.kg-1] resulted in a prolonged decrease in mean arterial blood pressure (MAP) and hyporeactivity to the contractile responses elicited by norepinephrine (NE; 10 nM) in aortic rings ex vivo. Hyporeactivity to NE was partially reversed by NG-nitro-L-arginine methyl ester (0.3 mM) in vitro, suggesting that an enhanced formation of NO contributes to this hyporeactivity. There was a substantial increase in the activity of iNOS in the lung 3 h after i.v. injection of LPS (0.2 +/- 0.1 to 6.6 +/- 0.6 pmol.mg-1.min-1; n = 5; P < 0.01). Rats injected i.p. with LPS (0.5 mg.kg-1) for 4 consecutive days became tolerant to an i.v. injection of LPS (2 mg.kg-1) in that both hypotension and vascular hyporeactivity to NE were significantly attenuated. Moreover, in these endotoxin-tolerant rats, the induction of iNOS by LPS in the lung was attenuated by 63% +/- 6%. Injection of LPS caused a 9-fold increase in plasma corticosterone (CCS) levels within 2 h and CCS levels remained significantly elevated 6 and 24 h after LPS. Animals rendered tolerant to endotoxin by administration of a low dose of LPS (0.5 mg.kg-1, i.p.) for 4 days still had a 6-fold increase in plasma CCS levels 24 h after the last injection of LPS. When endotoxin-tolerant rats were treated with the glucocorticoid receptor antagonist RU 486 (50 mg.kg-1, p.o. 3 h prior to LPS), there was a restoration of the effects of LPS (2 mg.kg-1, i.v.) in causing hypotension, vascular hyporeactivity to NE, and iNOS induction in the lung. However, in control rats RU 486 enhanced neither the decrease in MAP nor the induction of iNOS in response to LPS (2 mg.kg-1, i.v.). Thus, cardiovascular tolerance to endotoxin is accompanied and explained by reduced induction of iNOS in vivo due to the elevation of endogenous glucocorticoid levels.

Amino Acid Oxidoreductases↗

PubMed1994

The effects of the endothelin ETA receptor antagonist, FR 139317, on infarct size in a rabbit model of acute myocardial ischaemia and reperfusion.

1. The effects were investigated of the ETA receptor antagonist, FR 139317, on endothelin-1 (ET-1)-induced coronary vasoconstriction in the isolated perfused heart of the rabbit. In addition, this study examined whether FR 139317 reduced infarct size in a rabbit model of coronary artery occlusion and reperfusion. 2. In the rabbit isolated perfused heart, ET-1 (1-100 pmol) elicited a dose-dependent increase in coronary perfusion pressure (CPP). For example, 30 pmol ET-1 caused CPP to rise by 22 +/- 8 mmHg and 100 pmol ET-1 by 47 +/- 10 mmHg (n = 8). Infusion of FR 139317 (1 microM) significantly attenuated the increase in CPP caused by ET-1 (30 pmol: 3 +/- 1 mmHg, 100 pmol: 8 +/- 2 mmHg; n = 8). 3. In the anaesthetized rabbit, infarct size (expressed as a percentage of the area at risk) after 45 or 60 min of coronary artery occlusion followed by 2 h of reperfusion was 47 +/- 6% (n = 6) and 55 +/- 7% (n = 5), respectively. A continuous infusion of FR 139317 (0.2 mg kg-1 min-1 preceded by a loading dose of 1.0 mg kg-1, i.v.; n = 5-6) had no effect on the extent of the myocardial infarct size (45 min: 47 +/- 6%; 60 min: 49 +/- 7%). Even a three-times higher dose (0.6 mg kg-1 min-1 preceded by a loading dose of 3 mg kg-1, i.v.; n = 4) of FR 139317 had no effect on myocardial infarct size (48 +/- 5%) after 45 min occlusion of the antero-lateral branch of the left coronary artery (LAL) and 2 h reperfusion.4. In a separate group of experiments, the LAL was occluded for 60 min and subsequently reperfused for 6 h. FR 139317 (0.6 mg kg-1 min-1 preceded by a loading dose of 3 mg kg-1, i.v.; n =4) had no significant effect on infarct size even in this long reperfusion model (control: 48 +/- 3%, FR 139317:61 +/- 6%).5. Thus, the vasoconstrictor effects elicited by ET-1 in the coronary vasculature of the rabbit are primarily mediated via the ETA receptor, for they were inhibited by the ETA receptor antagonist, FR 139317. However, an enhanced formation of endogenous ET-1 does not play a major role in ischaemia/reperfusion injury of the rabbit heart, for FR 139317 had no effect on infarct size.

Anesthesia↗

PubMed1994

Reversal of established responses to endothelin-1 in vivo and in vitro by the endothelin receptor antagonists, BQ-123 and PD 145065.

1. Endothelin-1 binds almost irreversibly to its receptors and causes prolonged vasoconstrictions. Here we have studied the reversal of established responses to ET-1 in vivo and in vitro by BQ-123, an ETA receptor-selective antagonist, and/or PD 145065, an ETA/ETB receptor non-selective antagonist. 2. In anaesthetized rats pretreated with hexamethonium, infusion of ET-1 (10(-11) mol kg-1 min-1) increased the mean arterial pressure (MAP) from 93 +/- 1.5 mmHg to 137 +/- 2.4 mmHg after 70 min (n = 29). While the ET-1 infusion was continued an additional infusion of BQ-123 caused a gradual dose-dependent reduction in the pressor effect of ET-1. For instance, after a 60 min infusion of BQ-123 (10(-8) mol kg-1 min-1) the MAP was decreased by 29.3 +/- 4.3 mmHg (n = 4). 3. PD 145065 was a much weaker antagonist of the established pressor effects of ET-1. At 10(-8) mol kg-1 min-1 it had no significant effect and even at 10(-7) mol kg-1 min-1 the elevated blood pressure was only reduced by 11.8 +/- 8.0 mmHg (n = 5) after 60 min. Co-infusion of BQ-123 and PD 145065 caused smaller reductions in the established response to ET-1 than infusion of BQ-123 alone. 4. Sustained contractions of rat aortic rings induced by ET-1 (3 x 10(-9) M) and mediated by ETA receptors were slowly reversed by addition of BQ-123 (10(-5) M) or PD 145065 (10(-5) M). For instance,after 40 min the elevated tone was reduced 85.8 +/- 5.6% (n = 6) by PD 145065, and 77.1 +/- 6.7% (n = 6)by BQ-123. Thus, on the rat aortic rings in vitro both antagonists were equally effective against established responses to ET-1.5. ET-1 increased the perfusion pressure of the rat isolated perfused kidney by 138.1 +/- 7.6 mmHg(n = 14). Subsequent co-infusion of BQ-123 or PD 145065 reversed this increase with PD 145065 being more active. For instance, PD 145065 (10-6 M) reversed the increase in perfusion pressure by 56.9 +/- 8.8% (n = 5) and BQ-123 (10-6 M) reversed it by 22.8 +/- 8.0% (n = 5). This fits well with the vasoconstriction induced by ET-1 in the rat kidney being mediated by ETA and ETB receptors.6. Thus, sustained vasoconstrictions to ET-1 in vitro, mediated by either ETA or ETB receptors, may be reversed slowly by the subsequent application of receptor antagonists. Similarly, endothelin antagonists reverse the pressor effects of ET-1 in vivo although co-antagonism of ETA and ETB receptors or the co-administration of an ETA receptor antagonist, BQ-123, and a mixed antagonist, PD 145065 produces less reversal than the application of an ETA receptor-selective antagonist. This may be because PD 145065 also reduces vasodilatations induced by ET-1 in vivo, or could suggest that because of its peptide structure PD 145065 affects the elimination of ET-1.

Amino Acid Sequence↗

PubMed1994

The effect of nitric oxide synthase inhibition on the plasma fibrinolytic system in septic shock in rats.

1. We have investigated the effect of pretreatment of rats with nitric oxide (NO) synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME) on the E. coli lipopolysaccharide (LPS)-induced changes in the plasma fibrinolytic system, platelet count, fibrinogen level, as well as in gross and microscopic pathophysiological changes indicative of disseminated intravascular coagulation (DIC) in rats. 2. E. coli LPS (6 mg kg-1, i.p.) produced a decrease in the levels of plasma fibrinogen and a drop in the blood platelet count 6 h after administration. The decrease in fibrinogen but not the drop in platelet count was reversed by pretreatment with L-NAME (30 mg kg-1, i.p., 24 h and 15 min before administration of LPS). 3. Pretreatment with L-NAME antagonized the LPS-induced activation of fibrinolysis as measured by changes in the euglobulin clot lysis time (ECLT) and enhanced the LPS-induced rise in the plasma level of plasminogen activator inhibitor (PAI). In animals pretreated with L-NAME there was also a marked reduction in the histological changes indicative of DIC. 4. We propose that L-NAME can act as a protective agent in LPS-induced DIC, and this protection is due to an increased generation of PAI following inhibition of NO synthase.

Amino Acid Oxidoreductases↗

PubMed1994

Inhibition by spermine of the induction of nitric oxide synthase in J774.2 macrophages: requirement of a serum factor.

Polyamines are endogenous regulators of various cell functions. Nitric oxide (NO) is a cytostatic and cytotoxic free radical which is produced by the inducible NO synthase (iNOS) in immuno-stimulated macrophages. We tested whether spermine modulates the induction of iNOS in J774.2 macrophages. Stimulation of macrophages by bacterial lipopolysaccharide (LPS) or gamma-interferon increased the accumulation of nitrite in the culture medium. Spermine (10(-6) - 10(-4) M) inhibited nitrite production without causing cytotoxicity. This inhibition of NO formation by spermine was significantly reduced when it was given 6 h after LPS. Spermine did not inhibit nitrite accumulation when foetal calf serum was omitted from the tissue culture medium. Thus, spermine is an inhibitor of the induction of iNOS, and its inhibitory activity requires the presence of a serum factor.

Amino Acid Oxidoreductases↗

PubMed1994

Characterization of endothelin (ET) receptors in the isolated gall bladder of the guinea-pig: evidence for an additional ET receptor subtype.

1. We have characterized the receptors mediating contractions induced by endothelin-1 (ET-1), ET-2, ET-3 and the ETB-selective receptor agonists, sarafotoxin 6c (SX6c), IRL 1620, BQ-3020, [Ala1,3,11,15]ET-1 and ET (16-21) in strips of the isolated gall bladder of the guinea-pig (GPGB). We used as antagonists BQ-123 (ETA receptor selective) and PD 145065 (ETA/ETB receptor non-selective). 2. ET-1, ET-2 and ET-3 (10(-10) M to 3 x 10(-7) M) caused similar slowly-developing concentration-dependent contractions of the GPGB. Contractile effects induced by ET-1, ET-2 or ET-3 (at 3 x 10(-7) M) were also similar (230 +/- 25, 241 +/- 7 and 287 +/- 37% of that to histamine at 5 x 10(-6) M, n = 7, 6, 12, respectively). However, the threshold concentration for ET-1 or ET-2 was 10(-10) M whereas it was 3 x 10(-9) M for ET-3. 3. SX6c (10(-10) M to 3 x 10(-7) M) also caused slowly-developing concentration-dependent contractions at a threshold concentration of 10(-10) M (n = 16). However, the contraction caused by SX6c at 3 x 10(-7) M was 116 +/- 9% of that to histamine at 5 x 10(-6) M, which was half of that induced by the same concentration of the ET isopeptides. The contraction induced by IRL 1620 at 3 x 10-7 M (n = 9) was 43 +/- 9% of that to histamine at 5 x 10-6 M, which was one fifth of that produced by the same concentration of ET-1. Contractions induced by BQ-3020 or [Ala1,3,11,15]ET-I at 3 x 10-7 M were even less than those produced by IRL 1620. ET (16-21) was inactive up to 10-5 M. Addition of a concentration of 3 x 10-7 M of ET-1 to tissues with developed contractions induced by the bolus addition of 3 x 10-7 M SX6c caused a further contraction of the GPGB to the level observed with ET-1 alone at 3 x 10-7M (n = 8).4. BQ-123 (10-5 M) did not affect the concentration-response curve to ET-1 and the contraction induced by 3 x 10- M was also not affected (n = 5; 239 +/- 19% of histamine at 5 x 10-6 M). PD 145065(10-5 M) shifted the ET-1 concentration-response curve to the right and the contraction induced by ET-1at 3 x 10-7 M was inhibited by 15% (n = 6; NS). A higher concentration of BQ-123 (10-4 M) caused a significant shift to the right of the ET-1 concentration-response curve similar to that caused by PD 145065 (10-s M) and caused a 24% (n = 6; NS) inhibition of the contractions induced by ET-1 at 3 x 10-7 M. PD 145065 (10-4 M) abolished contractions induced by ET-1 (up to 10- M) and inhibited the response to ET-1 at 3 x 10-7 M by 52% (n = 4; P< 0.05).5. Contractions induced by ET-3 were more sensitive to inhibition by the antagonists. BQ-123 (10-6,10-5 or 10-4 M) inhibited responses to 3 x 10-7 M ET-3 by 66, 71 and 83%, respectively (n = 5, 5, 3;P< 0.05). PD 145065 (10-6, 10-5 or 10-4 M) attenuated more strongly than did BQ-123 the contractions induced by ET-3. For instance, the contractions caused by ET-3 at 3 x 10-7 M were decreased by 73 and 80% (n = 5, 5; P<0.05) in the presence of PD 145065 (10-6 or 10-5 M, respectively). PD 145065(10-4 M) completely abolished contractions to ET-3 (n = 4; up to 3 x 10-7M).6. Contractions induced by SX6c, especially those observed at concentrations lower than 10-8 M, were attenuated by BQ-123 (up to 10-4 M). PD 145065 (10-5M) shifted to the right the concentration response curve to SX6c and inhibited by 38% (P<0.05) the contractions induced by 3 x 10-7M.However, the contractions induced by a bolus addition of a high concentration of SX6c (3 x 10-7 M)and the subsequent addition of an identical concentration of ET-1 on top of SX6c were not affected byBQ-123 (10-6 or 10-5 M).7. These results suggest that ETB receptors are involved in the contractions induced by endothelins in the GPGB. However, SX6c and other selective ETB agonists produced only half or less than half of the contractile response induced by non-selective agonists. In addition, the responses to ET-1 but not to ET-3, were insensitive to the antagonist action of BQ-123 at 10-5 M whereas BQ-123 or PD 145065 at 10-5 M strongly antagonized contractions induced by ET-3. Finally, BQ-123 at 10-4 M inhibited contractions to ET-1 and SX6c. Thus, within the GPGB there may well be additional ET receptor(s) not conforming to the established ETA/ETB receptor subtype classification, as well as ETB receptors.

Amino Acid Sequence↗

PubMed1994

5-HT3 receptors do not mediate vagally-induced relaxation or contraction of the isolated stomach of the guinea-pig.

1. We have tested whether 5-HT3 receptors mediate vagally-induced relaxation or contraction of the isolated stomach of the guinea-pig. 2. The antagonists of 5-HT3 receptors, ondansetron (1-10 microM) or metoclopramide (1-30 microM) did not inhibit vagally-induced relaxations at low concentrations but partially inhibited them at 30 microM or 60 microM, respectively. These higher concentrations of ondansetron and metoclopramide also inhibited relaxations induced by 1,1-dimethyl-4-phenylpiperazinium iodide (30 microM) but did not affect those induced by glyceryl trinitrate (0.7-1.1 microM). 3. Desensitization to 5-HT (100 microM) or 2-Me-5-HT (100 microM) did not affect relaxations or contractions induced by vagal stimulation. 4. Ondansetron (30 microM) or metoclopramide (60 microM) did not inhibit vagally-induced gastric contraction. 5. Thus, 5-HT3 receptors do not mediate vagally-induced relaxation or contraction in the guinea-pig stomach.

Animals↗

PubMed1994

Evidence from receptor antagonists of an important role for ETB receptor-mediated vasoconstrictor effects of endothelin-1 in the rat kidney.

1. To characterize the receptor subtype(s) mediating the renal vasoconstrictor effects of the endothelin (ET) and sarafotoxin (SX) peptides in the isolated perfused kidney of the rat, we have examined the effects of endothelin-1 (ET-1), sarafotoxin 6b (SX6b) and sarafotoxin 6c (SX6c) as agonists, BQ-123 and FR 139317 as selective ETA receptor antagonists, and PD 145065 as a non-selective (ETA and ETB) receptor antagonist. We have also compared in the anaesthetized rat the systemic pressor and renal vasoconstrictor effects of ET-1 and SX6c alone or after pretreatment with PD 145065. 2. In the isolated perfused kidney, ET-1, SX6b and SX6c all gave similar concentration-dependent increases in perfusion pressure. The ETA receptor selective antagonists, BQ-123 and FR 139317, both partially blocked the increase in perfusion pressure induced by ET-1. In contrast, PD 145065 completely blocked the increase in perfusion pressure caused by ET-1. 3. Indomethacin (10 microM) had no effect on the ET-1-induced increases in perfusion pressure but significantly reduced the vasoconstriction induced by low concentrations of SX6c, without affecting responses to high concentrations. In the anaesthetized rat, indomethacin (5 mg kg-1) did not modify the systemic pressor or renal vasoconstrictor effects of ET-1 or SX6c. 4. In anaesthetized rats, bolus intravenous injections of ET-1 or SX6c (0.1, 0.25, 0.5 or 1.0 nmol kg-1) produced initial transient depressor responses followed by sustained and dose-dependent increases in mean arterial pressure (MAP). Both peptides caused an equipotent fall in renal blood flow (RBF).PD 145065 (5 mg kg-1) partially antagonized the systemic pressor effects of ET-1 and SX6c but completely blocked the fall in RBF and rise in renal vascular resistance (RVR) induced by ET-1 and SX6c. PD 145065 also antagonized the transient depressor effect following the bolus administration of either ET-1 or SX6c.5. These results indicate that ET/SX induced renal vasoconstriction is mediated via ETA and ETB-like receptors with ETB receptors having a predominant role in vivo. This may be of therapeutic relevance for an ETA receptor-selective antagonist may offer only limited protection against the deleterious renal effects of endogenous ETs.

Amino Acid Sequence↗

PubMed1994

Characterization of ETB receptors mediating contractions induced by endothelin-1 or IRL 1620 in guinea-pig isolated airways: effects of BQ-123, FR139317 or PD 145065.

1. We have characterized the receptors mediating contractions to endothelin-1 (ET-1) or IRL 1620, an ETB receptor selective agonist, in isolated strips of tissue prepared from different parts of the guinea-pig airways. We used as antagonists BQ-123 and FR139317 (ETA receptor-selective) and PD 145065 (ETA/ETB receptor non-selective). 2. ET-1 and IRL 1620 (10(-10) M to 10(-6) M) caused similar concentration-dependent contractions of strips of guinea-pig trachea and upper bronchus. In the guinea-pig trachea without epithelium or lung parenchyma, IRL 1620 was less potent than ET-1. 3. In the trachea, contraction to ET-1 (< 10(-8) M) was preceded by a transient relaxation which was inhibited by BQ-123 (10(-5) M) or FR 139317 (10(-5) M) or by the removal of the epithelium. The concentration-response curve to ET-1 in the trachea was shifted to the right by PD 145065 (10(-5) M to 10(-4) M). PD 145065 (10(-4) M) also inhibited the response to ET-1 (3 x 10(-7) M) by 55%. Contractions induced by IRL 1620 were not affected by BQ-123 (10(-6) M) or FR139317 (10(-6) M) but were significantly attenuated by 10(-5) M of either antagonist. PD 145065 at 10(-6) M strongly attenuated and at 10(-5) M abolished contractions induced by IRL 1620. 4. In the trachea, removal of the epithelium potentiated the effects of both agonists. BQ-123 (10-5 M)had no effect on contractions of the trachea without epithelium induced by ET-1, but FR139317 (10-5 M)caused a significant inhibition. PD 145065 (10-5 M to 10-4 M) caused a shift to the right of the ET-1 concentration-response curve without affecting the contractile effect at 3 x 10-7 M. All three antagonists inhibited contractions induced by IRL 1620.5. In the upper bronchus, BQ-123 (10-5 M) did not affect contractions induced by ET-1, whileFR139317 (10-5 M) attenuated (20-26%) only contractions induced by 1-3 x 10-7 M ET-1. PD 145065(10-5 M to 10-4 M) caused a shift to the right of the ET-1 concentration-response curve. The contractions induced by IRL 1620 were inhibited by BQ-123 or FR139317 (10-5M to 10-4 M). PD 145065(10-6 M) strongly inhibited contractions induced by IRL 1620 and PD 145065 (10-5 M) totally abolished them.6. The contractile action of ET-1 in the lung parenchyma was significantly and similarly attenuated by BQ-123 (10-5 M) or indomethacin (10-5 M), while FRI39317 (10-5 M) was less effective. PD 145065(10-6 to 10-5 M) inhibited contractions to ET-1. IRL 1620, which is less potent than ET-1 in this preparation, was antagonized by PD 145065 (10-5 to 10-6 M) but unaffected by BQ-123 (10-6 M to10-5M) or FR139317 (10-6 M).7. Thus, ETB receptors mediate contractions to ET-1 in all four guinea-pig airway preparations. In addition, contractions to ET-1 in the trachea and lung parenchyma are mediated in part by ETA receptors. In the latter tissue, these ETA receptors mediate contraction through the release of cyclooxygenase metabolites. Similarly, ETA receptors located on the epithelial cells also mediate the release of prostanoids in the trachea with epithelium but they are responsible for transient relaxations. Interestingly,contractions induced by IRL 1620 were more susceptible to inhibition by the different antagonists,most probably because it binds to the endothelin receptors in a reversible manner. High concentrations(10-5 M) of ETA-selective antagonists also inhibit responses to IRL 1620, most probably by an effect at ETB receptors in both the trachea and the upper bronchus.

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