endothelin-1 and cicletanine

Cicletanine (CIC), an anti-hypertensive compound with direct vascular and natriuretic actions, is especially effective in salt-sensitive hypertension, in which dysregulation of the sodium pump plays an important pathogenic role, and digitalis-like cardiotonic steroids contribute to increased vascular tone. The purpose of the present study was to investigate whether, and by what mechanisms, cicletanine antagonizes the vasoconstrictor effects of cardiotonic steroids in isolated human arteries.. The effects of cicletanine on vascular tone were studied in isolated, endothelium-denuded rings of 2nd-3rd-order branches of human mesenteric arteries pre-contracted with bufodienolide marinobufagenin (MBG), an Na/K-ATPase inhibitor, or endothelin-1 (ET-1). Na/K-ATPase activity was measured in sarcolemmal membranes from the mesenteric artery. Activity of rat brain protein kinase C (PKC) was measured using the PepTag phosphorylation assay.. MBG and ET-1 both induced sustained vasoconstriction in human mesenteric artery rings, and cicletanine relaxed rings pre-contracted with either MBG (EC50 = 11 +/- 2 micromol/l) or ET-1 (EC50 = 6.4 +/- 1.1 micromol/l). Although 8-Br-cGMP (100 micromol/l) caused complete vasorelaxation of arterial rings pre-contracted with ET-1, it did not affect the MBG-induced vasoconstriction. An activator of PKC, phorbol diacetate (PDA) (50 nmol/l), attenuated CIC-induced vasorelaxation of mesenteric artery rings pre-contracted with MBG (EC50 > 100 micromol/l), but not rings pre-contracted with ET-1 (EC50 = 6.5 +/- 1.2 micromol/l). In mesenteric artery sarcolemma, 100 nmol/l MBG inhibited the Na/K-ATPase by 68 +/- 5% and cicletanine (100 micromol/l) attenuated this Na/K-ATPase inhibition by 85 +/- 6%. In the PepTag PKC assay, cicletanine produced a concentration-dependent inhibition of rat brain PKC activity (IC50 45 +/- 11 micromol/l). In the presence of 50 nmol/l PDA, 100 micromol/l cicletanine did not antagonize the Na/K-ATPase inhibition by MBG, and did not inhibit the PKC from rat brain.. Cicletanine antagonizes vasoconstriction induced by Na/K-ATPase inhibition via a PKC-dependent mechanism that does not involve inhibition of cyclic GMP phosphodiesterase (cGMP-PDE). This mechanism of action may be relevant to the greater potency of cicletanine in salt-sensitive hypertension in which plasma levels of endogenous digitalis-like cardiotonic steroids are elevated. Our findings also suggest that PKC is an important factor for cardiotonic steroid-Na/K-ATPase interactions on the vascular tone, and is therefore a potential target for therapeutic intervention in hypertension.

Topics: Animals; Antihypertensive Agents; Brain; Bufanolides; Cyclic GMP; Endothelin-1; Enzyme Inhibitors; Humans; In Vitro Techniques; Male; Mesenteric Arteries; Middle Aged; Protein Kinase C; Pyridines; Rats; Sodium-Potassium-Exchanging ATPase; Vasoconstriction

The purpose of the study was to investigate, in isolated human pulmonary artery, the ability of cicletanine and its (-) and (+)-enantiomers to attenuate the endothelin-1 (Et-1) induced vasoconstriction, and to potentiate vasorelaxation (relative to plateau of the effect of Et-1) by sodium nitroprusside (SNP) and human atrial natriuretic peptide (ANP). In pulmonary artery rings, Et-1 induced a concentration-dependent vasoconstriction with median effective concentration (EC50 = 26+/-2.8 nmol/L. Pretreatment of the vessels with 100 micromol/L (+/-)-cicletanine reduced the effect of Et-1 (EC50 = 36+/-3.5 nmol/L; P < .01). (-)-enantiomer displayed greater capacity to antagonize the vasoconstrictor action of Et-1 (EC50 = 47+/-4.2 nmol/L) v (+)-enantiomer (EC50 = 29.9+/-6.5 nmol/L; P < .01). In arterial rings, precontracted with 10 nmol/L Et-1, ANP caused vasorelaxation (EC50 = 9.7+/-1.9 nmol/ L). The relaxant effect of ANP was potentiated by 100 micromol/L of (-)-(EC50 = 4.2+/-0.6 nmol/L; P < .01), but not (+)-cicletanine (EC50 = 7.6+/-0.7 nmol/L). Sodium nitroprusside relaxed pulmonary artery rings precontracted with 10 nmol/L Et-1 (EC50 = 41+/-11 nmol/L). The effect of SNP was potentiated by 10 micromol/L (+/-)-cicletanine (EC50 = 9.0+/-0.7 nmol/L; P < .05). The potentiating effect of 10 micromol/L (+)-cicletanine was weaker (EC50 = 7.9+/-1.8 nmol/L) than that of (-)-enantiomer (EC50 = 3.3+/-0.54 nmol/L; P < .05). The relaxant effect of SNP was not further potentiated by 100 micromol/L (+/-)-cicletanine. The present results demonstrate that, cicletanine antagonizes Et-1 induced vasoconstriction in an isolated human pulmonary artery and potentiates vasorelaxation by two guanylate cyclase activators, ANP and SNP. (-)-Cicletanine displays greater vasorelaxant activity v (+)-enantiomer.

Topics: Adult; Antihypertensive Agents; Atrial Natriuretic Factor; Dose-Response Relationship, Drug; Drug Synergism; Endothelin-1; Humans; In Vitro Techniques; Male; Middle Aged; Nitroprusside; Pulmonary Artery; Pyridines; Stereoisomerism; Vasodilator Agents