s6c-sarafotoxin and 3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate

1. The isolated superior mesenteric arterial bed of the rat, perfused with Krebs-Henseleit solution containing 10 microM indomethacin, was used to study the effects of reducing dissolved O2 tension on the pressor responses to endothelin-1, endothelin-3 and sarafotoxin S6b. The modulation of these responses by the endothelium was investigated by removing the intima with the detergent CHAPS and, for endothelin-1, by inhibiting nitric oxide production with N omega-nitro-L-arginine methyl ester (L-NAME). Comparison was made with the effects of lowering O2 tension on the pressor responses to noradrenaline and 5-hydroxytryptamine. 2. Lowering the perfusate O2 tension from 551 +/- 2 mmHg to 14.0 +/- 0.5 mmHg did not change the ED50 for endothelin-1 but its maximal responses (Rmax) were increased by 2.1 and 2.7 fold, respectively, in the presence and absence of endothelium. The Rmax values for endothelin-3 were also greater in hypoxia either in the presence (by 2.3 fold) or absence of the endothelium (by 1.6 times) but those for sarafotoxin S6b were only enhanced significantly by hypoxia in the absence of the intima. hypoxia reduced the potencies of endothelin-3 and sarafotoxin S6b whether or not endothelium was present. 3. Endothelial destruction, whether in hypoxic or oxygenated conditions, increased the Rmax values for endothelin-1 and endothelin-3; at both O2 tensions those for endothelin-3 increased more than those for endothelin-1. The ED50 for endothelin-1 was unchanged by destroying the endothelium but endothelin-3 was less potent in the absence of an endothelium than in its presence. Removal of the endothelium did not change the R.ax for sarafotoxin S6b but increased its potency in both hypoxic and oxygenated tissues. 4. In hypoxia, and in the presence of both the endothelium and 100 microM L-NAME, the Rmax for endothelin-1 was 1.6 times greater than that in hypoxia in the absence of L-NAME. Co-infusion of 100 microM L-arginine, but not of 100 mircoM D-arginine, with 100 microM L-NAME reversed this effect. The presence of L-NAME decreased the potency of endothelin-1. 5. Destroying the endothelium did not affect the Rmax for noradrenaline in either oxygenated conditions or hypoxia. Changing 02 tension when the endothelium was intact had no effect on the Rmax but it was 11% greater in oxygenated, than in hypoxic, endothelium denuded preparations. Endothelial destruction decreased the potency of noradrenaline in hypoxia but increased it in oxygenated tissue

Topics: Animals; Arginine; Blood Pressure; Cholic Acids; Endothelins; Endothelium, Vascular; Hypoxia; In Vitro Techniques; Male; Mesenteric Arteries; Muscle, Smooth, Vascular; NG-Nitroarginine Methyl Ester; Norepinephrine; Oxygen Consumption; Rats; Rats, Inbred Strains; Vasoconstrictor Agents; Viper Venoms

1. The vasoconstrictor activities of endothelin-2, endothelin-3, sarafotoxin S6b, human proendothelin1-38 and mouse vasoactive intestinal contractor (VIC) were studied in the isolated Krebs-Henseleit perfused mesenteric arterial bed of the rat in the presence and absence of the endothelium. The vasoconstrictor properties of endothelin-1 were studied in control preparations and in preparations treated with methylene blue or N omega-nitro-L-arginine methyl ester (NAME). Finally, the direct vasodilator properties of endothelin-2, endothelin-3 and sarafotoxin S6b were studied in preparations preconstricted with methoxamine. 2. In the presence of an intact endothelium, all of the peptides caused dose-dependent increases in perfusion pressure and sarafotoxin S6b was a full agonist relative to the other peptides studied (maximum increase in perfusion pressure, Rmax = 106 +/- 11 mmHg). Endothelin-1, endothelin-2 and VIC were more potent vasoconstrictors (ED50 93.0 +/- 40.0, 90.8 +/- 20.5 and 106 +/- 63 pmol, respectively) than endothelin-3 and sarafotoxin S6b, which were found to be equipotent (ED50 values 411 +/- 195 and 345 +/- 86 pmol, respectively). A full dose-response relationship could not be constructed for proendothelin, but the highest dose used (4 nmol) increased the perfusion pressure by 15.4 +/- 1.6 mmHg. 3. Destruction of the endothelium with the zwitterionic detergent 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS) significantly enhanced the pressor activity of all 5 peptides. The Rmax for sarafotoxin S6b was not significantly altered by removal of the endothelium but its potency was significantly increased (ED50 = 115 +/- 15 pmol). Although their R,,, values were significantly increased, endothelin-2 and VIC were still partial agonists relative to sarafotoxin S6b in CHAPSpretreated preparations; their potencies were unchanged (ED5o values 118 + 53 and 416 + 196pmol, respectively). Removal of the endothelium significantly reduced the potency of endothelin-3 (ED5o, 6.3 + 2.2 nmol) but this peptide then exhibited full agonist activity (R..x = 106 + 14 mmHg). After endothelial cell destruction, the pressor responses to proendothelin were increased; 4 nmol gave a response of 38.8 + 5.5 mmHg. 4. Exposure of preparations to either 100 microM NAME (R,,,X = 42.6 + 2.4mmHg and EDSo = 57.5 + 13.7 pmol) or 10 microM methylene blue (R,,,. = 36.0 + 5.1 mmHg and ED50 = 81.5 + 26.1 pmol) significantly enhanced the maximum pressor responses to

Topics: Animals; Arginine; Blood Pressure; Cholic Acids; Endothelins; Endothelium, Vascular; In Vitro Techniques; Male; Mesenteric Arteries; Methylene Blue; Muscle Relaxation; Muscle, Smooth, Vascular; Nitric Oxide; Nitroarginine; Rats; Rats, Inbred Strains; Vasoconstriction; Viper Venoms