title: The effect of endothelins on nitric oxide and prostacyclin production from human umbilical vein, porcine aorta and bovine carotid artery endothelial cells in culture.
abstract: 1. This study has investigated the effects of the endothelin isopeptides, endothelin-1 (ET-1), ET-2 and ET-3 on the production of the endothelium-derived relaxing factors, nitric oxide (NO) and prostacyclin (PGI2) from primary cultures of endothelial cells obtained from human umbilical vein (HUVECS), porcine aorta (PAECS) and bovine carotid artery (BCAECS). 2. NO generation was assessed indirectly by measuring production of cyclic GMP and PGI2 formation was measured by radioimmunoassay of 6-keto PGF1 alpha. 3. In HUVECS, histamine (1 microM) increased cyclic GMP and 6-keto PGF1 alpha production by 12.6 +/- 2.0 and 4.9 +/- 0.7 fold respectively over the corresponding basal values. Haemoglobin (10 microM) and the NO synthase inhibitor NG-monomethyl-L-arginine (10 microM) significantly inhibited the increase in cyclic GMP formation in response to histamine but had no effect on 6-keto PGF1 alpha production. In contrast to histamine, the endothelin isopeptides (ET-1, ET-2 and ET-3; 0.01-1000 nM) produced no significant change in either cyclic GMP or 6-keto PGF1 alpha production in HUVECS. 4. In a separate series of experiments, ET-3 (0.01-1000 nM) also failed to produce any significant change in cyclic GMP or 6-keto PGF1 alpha production from primary cultures of PAECS and BCAECS. In contrast, bradykinin (0.1 microM) and sodium nitroprusside (1 mM) were used as positive control agents and increased cyclic GMP production in these cells. 5. In conclusion, the endothelin isopeptides do not release NO and PGI2 from primary cultures of HUVECS, PAECS and BCAECS. This suggests that endothelin receptors are either absent from these cells or are not coupled to NO or PGI2 production.
authors.affiliation: Department of Cardiovascular and Respiratory Pharmacology, Glaxo Group Research Ltd., Ware, Hertfordshire.
authors.forename: D G
authors.initials: DG
authors.lastname: White
authors.forename: J W
authors.initials: JW
authors.lastname: Mundin
authors.forename: M J
authors.initials: MJ
authors.lastname: Sumner
authors.forename: I S
authors.initials: IS
authors.lastname: Watts
mesh_terms: D015121:6-Ketoprostaglandin F1 alpha / Q000096:biosynthesis; D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D001920:Bradykinin / Q000494:pharmacology; D002339:Carotid Arteries / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D016232:Endothelins / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D011464:Epoprostenol / Q000096:biosynthesis*; D005260:Female; D006454:Hemoglobins / Q000494:pharmacology; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D009569:Nitric Oxide / Q000096:biosynthesis*; D009599:Nitroprusside / Q000494:pharmacology; D011247:Pregnancy; D013552:Swine; D014471:Umbilical Veins / Q000187:drug effects / Q000378:metabolism; D019323:omega-N-Methylarginine
chemical_list: D016232:Endothelins; D006454:Hemoglobins; D009599:Nitroprusside; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D015121:6-Ketoprostaglandin F1 alpha; D006632:Histamine; D001120:Arginine; D011464:Epoprostenol; D006152:Cyclic GMP; D001920:Bradykinin