cyclic-gmp and nipradilol

Nitric oxide (NO) is cytoprotective to certain types of neuronal cells. The neuroprotective ability of NO in the retina was reportedly mediated by the cyclic GMP (cGMP) to protein kinase G (PKG) pathway. Cyclic AMP-responsive element binding protein (CREB) plays an essential role in the NO/cGMP/PKG-mediated survival of rat cerebellar granule cells. We tested whether CREB transduces the NO/cGMP/PKG anti-apoptotic cascade in R28 neuro-glial progenitor cells. Apoptosis was induced in R28 cells by serum deprivation for 24 h. Varying concentrations of two NO donors, sodium nitroprusside (SNP) and nipradilol, were added to medium with or without an NO scavenger, a soluble guanylyl cyclase inhibitor, or a PKG inhibitor. The cells were immunostained against activated caspase-3 and counterstained with Hoechst 33258. Apoptosis was quantified by counting activated caspase-3 positive or pyknotic cells. SNP and nipradilol rescued R28 cells from apoptosis in a dose-dependent manner, at an optimal concentration of 1.0 microM and 10 microM, respectively. Higher concentrations were cytotoxic. The NO scavenger and the inhibitors decreased the anti-apoptotic effect of the NO donors. Intracellular cGMP levels were increased after exposure to SNP and nipradilol. Western blotting showed that both NO donors increased CREB phosphorylation, which was blocked when pre-exposed to the inhibitors. Transfection with a dominant negative CREB construct defective of phosphorylation at Ser-133 interfered with the anti-apoptotic activity of SNP. These results indicate that CREB at least in part mediates the cGMP/PKG-dependent anti-apoptotic signal induced by NO in R28 cells.

Topics: Animals; Apoptosis; Cells, Cultured; Cyclic AMP Response Element-Binding Protein; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Cytoprotection; Dose-Response Relationship, Drug; Neuroglia; Nitric Oxide; Nitric Oxide Donors; Nitroprusside; Phosphorylation; Propanolamines; Rats; Retina; Signal Transduction; Stem Cells

Nipradilol (3,4-dihydro-8-(2-hydroxy-3-isopropylamino)propoxy-3-nitroxy-2H-1-benzopyran) is used clinically as an anti-glaucoma ophthalmic solution in Japan, and was recently reported to suppress N-methyl-d-aspartate-induced retinal damage in rats. Here we investigated cytotoxic and cytoprotective actions of nipradilol on primary cultures of rat cortical neurons. Treatment of cortical cultures with a high concentration (500 microM) of nipradilol significantly reduced cell viability, increased lactate dehydrogenase (LDH) release and nitrite concentration in culture medium, whereas desnitro-nipradilol (3,4-dihydro-8-(2-hydroxy-3-isopropylamino)propoxy-3-hydroxy-2H-1-benzopyran) had no significant effects. Nipradilol-induced neuronal damage was inhibited by S-hexylglutathione, a glutathione S-transferase inhibitor, and FeTPPS (5,10,15,20-tetrakis(4-sulfonatophenyl)prophyrinato iron (III) chloride), a peroxynitrite decomposition catalyst. On the other hand, relatively low concentrations (10-100 microM) of nipradilol but not desnitro-nipradilol prevented neuronal cell death induced by 24 h application of 100 microM glutamate. Importantly, neuroprotective concentration (100 microM) of nipradilol suppressed glutamate-induced elevation of intracellular Ca2+ concentrations, but had no effect on intracellular cyclic GMP levels. Hence, nipradilol can protect cultured cortical neurons against glutamate neurotoxicity via cyclic GMP-independent mechanisms, and nitric oxide (NO) released from the nitoroxy moiety of nipradilol may mediate neuroprotective effect through the modulation of NMDA receptor function.

Topics: Adrenergic alpha-Antagonists; Adrenergic beta-Antagonists; Animals; Calcium; Cell Survival; Cells, Cultured; Cerebral Cortex; Cyclic GMP; Dizocilpine Maleate; Dose-Response Relationship, Drug; Enzyme Inhibitors; Female; Fetus; Glutamic Acid; Glutathione; Lactoylglutathione Lyase; Neurons; Neuroprotective Agents; Nitric Oxide; Pregnancy; Propanolamines; Rats; Rats, Wistar

The L-arginine-nitric oxide (NO) pathway plays an important role in the modulation of glomerular disease. We investigated whether beta-blocking agents, with and without an NO-generating function, had renoprotective effects in the 5/6 nephrectomized rats (Nx), an animal model of glomerulosclerosis.. Nipradilol, a beta-blocker with an ONO(2) group (5, 10 or 15 mg/kg/day) and propranolol, a beta-blocker without this group (50 mg/kg/day) were administered for 12 weeks to Nx together with and without nitro-L-arginine methyl ester (L-NAME). We evaluated the effects of both drugs on proteinuria, hypertension, renal function, glomerulosclerosis and urinary excretion of NO metabolites (U(NOx)) and cyclic GMP (U(cGMP)).. Both drugs similarly attenuated the elevated blood pressure in Nx. However, nipradilol, at doses of 10 and 15 mg/kg/day, significantly decreased proteinuria and glomerulosclerosis, while propranolol did not. Nx showed reduced U(NOx) in comparison with the sham-operated rats. Nipradilol increased U(NOx) and U(cGMP) significantly and in a dose- dependent manner, whereas propranolol reduced them to levels lower than those in Nx. Nx receiving L-NAME reduced U(NOx). The addition of nipradilol increased U(NOx) and decreased urinary protein excretion and glomerulosclerosis, suggesting that the NO released from the drug contributed to its renoprotective effect.. These findings indicate that nipradilol exerts its renoprotective effect through NO generation, and not by lowering blood pressure. The beta-adrenergic blocking action per se does not seem to be related to the renoprotective effect of these agents.

Topics: Adrenergic beta-Antagonists; Animals; Cyclic GMP; Enzyme Inhibitors; Hypertension; Kidney; Kidney Diseases; Kidney Glomerulus; Male; Nephrectomy; NG-Nitroarginine Methyl Ester; Nitrates; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase; Nitrites; Propanolamines; Propranolol; Proteinuria; Rats; Rats, Inbred F344; Renin

To study whether nipradilol, which is used as an ophthalmic solution for the treatment of glaucoma, has a cytoprotective effect, we investigated its effect on the apoptosis induced by serum withdrawal in PC12 cells. Nipradilol has alpha1- and beta-adrenoceptor-blocking and nitric oxide (NO)-donating properties. We also investigated the effects of timolol, prazosin and S-nitroso-N-acetylpenicillamine (SNAP) on PC12 cell death. Serum withdrawal from PC12 cells resulted in apoptosis, and the survival rate was decreased in a time-dependent manner. The addition of nipradilol to the medium showed a cytoprotective effect on PC12 cell death in a dose-dependent manner, but timolol and prazosin did not. We measured caspase-3 activity to clarify the mechanism of the inhibition of apoptosis in the presence or absence of dithiothreitol (DTT). The caspase-3 activity could be reactivated by DTT. In addition, to investigate the relationship of the cGMP-dependent pathway to the nipradilol-induced cytoprotective effect, we tested the effect of the protein kinase G inhibitor KT5823. KT5823 partially reversed the nipradilol-mediated cytoprotective effect. These results indicate that the cytoprotective effect of nipradilol in PC12 cell death was due to the caspase-3 inhibition mediated by NO-related S-nitrosylation and activation of protein kinase G.

Topics: Animals; Apoptosis; Caspase 3; Caspase Inhibitors; Caspases; Cell Survival; Cyclic GMP; Cytoprotection; Dose-Response Relationship, Drug; Enzyme Activation; Nitric Oxide; PC12 Cells; Propanolamines; Rats; Signal Transduction

There are reports of serious hypotension or circulatory shock when sildenafil citrate, a selective cyclic nucleotide phosphodiesterase type 5 inhibitor, which was developed for the treatment of erectile dysfunction, is given to patients taking certain coronary vasodilators. We thus examined the interaction of sildenafil with various coronary vasodilators including nitric oxide (NO) donors in isolated porcine coronary artery. Sildenafil caused concentration-dependent relaxations of the artery precontracted with U46619 (9,11-dideoxy-9 alpha,11 alpha-methanoepoxy-prostaglandin F(2alpha)). Incubation with the NO synthase inhibitor NG-nitro-L-arginine or the soluble guanylate cyclase inhibitor ODQ (1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one) significantly shifted the concentration-response curve for sildenafil to the right without affecting the maximum response, indicating that some part of the relaxant response to sildenafil may be the result of the inhibition of phosphodiestrase type 5-induced degradation of cyclic GMP (cGMP) that is produced through guanylate cyclase activation by NO released spontaneously. The relaxant effects of the vasodilators with an NO donor property, isosorbide dinitrate, sodium nitroprusside, nicorandil and nipradilol, were significantly enhanced by sildenafil, as shown by a significant leftward shift of their concentration-response curves. In contrast, the relaxant responses to the drugs without a property as an NO donor, diltiazem, celiprolol and pinacidil, were not affected by sildenafil. The cGMP level of the tissue was elevated after adding sildenafil, and the cGMP-generating effect of a combination of sildenafil and sodium nitroprusside was higher than that of each drug alone. The cyclic AMP level determined simultaneously was not changed by sildenafil. These results suggest that sildenafil potentiates specifically the relaxant responses of porcine coronary artery to the drugs which behave as an NO donor, providing basic evidence that the benefit of sildenafil in the treatment of erectile dysfunction can be limited by a risk of marked vasodilation when used together with NO-related coronary vasodilators.

Topics: Animals; Celiprolol; Coronary Vessels; Cyclic GMP; Diltiazem; Dose-Response Relationship, Drug; Drug Interactions; In Vitro Techniques; Nicorandil; Nitroprusside; Pinacidil; Piperazines; Propanolamines; Purines; Sildenafil Citrate; Sulfones; Swine; Vasodilation; Vasodilator Agents

We examined the effect of nipradilol on contraction of the posterior ciliary artery induced by high potassium or norepinephrine and on cyclic GMP (cGMP) levels in the posterior ciliary artery of dogs. Nipradilol caused dose-dependent relaxation of KCl-and norepinephrine-induced contractions of posterior ciliary artery. The relaxant effect of nipradilol on norepinephrine-contracted ciliary artery was significantly greater than that on KCl-contracted ciliary artery. In KCl-contracted ciliary artery, N(G)-nitro-L-arginine methyl ester hydrochloride (L-NAME, 10(-4) M) did not alter the relaxant effect of nipradilol, whereas 1H-1,2,4-oxadiazolo-4,3-a-quinoxalin-1-one (ODQ, 10(-6) M) significantly inhibited this effect. Ethacrynic acid at 10(-5) M, sulfasalazine at 10(-4) M and S-decylglutathione at 10(-4) M (glutathione S-transferase inhibitors) did not inhibit the relaxant effect of nipradilol. In addition, nipradilol produced dose-dependent increases in cGMP levels in the canine posterior ciliary artery. These findings indicate that nipradilol-induced vasorelaxation in the canine posterior ciliary artery occurs via stimulation of the guanylyl cyclase-cGMP pathway.

Topics: Adrenergic beta-Antagonists; Animals; Ciliary Arteries; Cyclic GMP; Dogs; Dose-Response Relationship, Drug; Enzyme Inhibitors; Glutathione Transferase; Guanylate Cyclase; In Vitro Techniques; Male; Muscle Contraction; Muscle Relaxation; Muscle, Smooth, Vascular; NG-Nitroarginine Methyl Ester; Nitric Oxide Synthase; Nitric Oxide Synthase Type III; Norepinephrine; Potassium; Propanolamines; Vasoconstrictor Agents; Vasodilation

We evaluated the effects of nipradilol, a beta-adrenoreceptor antagonist which contains a nitroxy residue, for vascular response in atherosclerosis of rabbits. Four groups of rabbits received different diets (standard diet; standard diet plus 10 mg/kg/day nipradilol; atherogenic diet [standard diet plus 1% cholesterol]; atherogenic diet plus 10 mg/kg/day nipradilol) for 9 weeks. Plasma lipids, blood pressure, vascular function, nitric oxide (NO), activity of NO synthase, cGMP, and histological atherosclerotic changes were evaluated. Neither the atherogenic diet nor nipradilol treatment affected significantly the animals' body weight, blood pressure, or heart rate. The atherogenic diet increased total cholesterol and triglycerides, which were not altered by nipradilol. The atherogenic diet diminished the acetylcholine-induced NO mediated relaxation. Nipradilol treatment restored this relaxation. Analyses using a NO-sensitive selective electrode showed that nipradilol released NO in the presence of cells and that NO release was greater in atherosclerotic aorta with than without nipradilol treatment. Nipradilol treatment increased the basal NO release as evaluated by the aortic tissue cyclic GMP (cGMP) levels in atherosclerotic vessel, and reduced the esterified cholesterol levels in atherosclerotic vessel. Conclusively, NO released by nipradilol may protect endothelium derived relaxation in atherosclerotic vessels, and may partially inhibit the accumulation of cholesterol in the atherosclerotic lesions.

Topics: Adrenergic beta-Antagonists; Animals; Arteriosclerosis; Cattle; Cells, Cultured; Cholesterol; Cyclic GMP; Endothelium, Vascular; Immunohistochemistry; Male; Muscle Relaxation; Nitric Oxide; Nitric Oxide Synthase; Propanolamines; Rabbits

1. Contribution of cyclic GMP generation to the relaxation by nipradilol of vascular smooth muscle was studied in the isolated rabbit aorta contracted by phenylephrine (10(-7) M) or prostaglandin F2 alpha (PGF2 alpha, 3 x 10(-6) M). 2. Nipradilol-induced relaxation in both contractions and increase in cyclic GMP content were inhibited by methylene blue (10(-5) M). 3. The relations between the increase in cyclic GMP and the relaxation produced by nipradilol and nitroglycerin were fitted to sigmoid curves. The increase in cyclic GMP at 50% relaxation by nipradilol was smaller than that by nitroglycerin (1.2-fold increase as against 3-fold increase). 4. These results suggest a smaller contribution of cyclic GMP generation through activation of soluble guanylate cyclase to nipradilol-induced relaxation in the rabbit aorta.

Topics: Adrenergic beta-Antagonists; Animals; Aorta; Cyclic GMP; In Vitro Techniques; Male; Nitroglycerin; Propanolamines; Rabbits; Vasodilation; Vasodilator Agents

Nipradilol, a beta-adrenoceptor antagonist with vasodilator activity, has a structure which contains a NO2 group. In anesthetized dogs, the time course of the systemic vasodilation and plasma nitrite (NO2-) concentration was studied following intravenous administration of nipradilol (1 mg/kg). A fall in systemic vascular resistance was observed at 1 min, which was rapidly followed by a significant increase in the plasma NO2- concentration. It is indicated that nipradilol exerts systemic vasodilatation via nitric oxide (NO) release in vivo.

Topics: Adrenergic beta-Antagonists; Animals; Atrial Natriuretic Factor; Catecholamines; Cyclic GMP; Dogs; Hemodynamics; Humans; Injections, Intravenous; Nitrites; Propanolamines; Vasodilator Agents

Nipradilol is a beta-adrenoreceptor blocking agent, whose structure contains an NO2 group. Thus, it is possible that it modulates the function of glomerular mesangial cells through the activation of soluble guanylate cyclase. To prove this hypothesis, we examined the effect of nipradilol on soluble guanylate cyclase, intracellular cyclic guanosine monophosphate (cGMP) accumulation, and the mitogenesis of cultured rat glomerular mesangial cells. Nipradilol increased intracellular cGMP accumulation in a dose-dependent manner through the activation of soluble guanylate cyclase. Furthermore, nipradilol inhibited the incorporation of [3H]thymidine into the mesangial cells stimulated by 2.5% fetal bovine serum in a dose-dependent manner. These results indicate that nipradilol may modulate mesangial cell function through an increase in intracellular cGMP resulting from the activation of soluble guanylate cyclase.

Topics: Adrenergic beta-Antagonists; Animals; Cell Division; Cells, Cultured; Cyclic GMP; Dose-Response Relationship, Drug; Enzyme Activation; Glomerular Mesangium; Guanylate Cyclase; Male; Propanolamines; Rats; Rats, Sprague-Dawley

The relaxant effect on smooth muscle of nitro compounds is suggested to be linked with the increase in the tissue level of cyclic GMP by activating guanylate cyclase. In this study, we investigated the effects of nipradilol, a new beta-blocker, which has NO2 residue in the molecular structure, and isosorbide dinitrate (ISDN) on guinea pig tracheal smooth muscle in comparison with the effect of propranolol. Nipradilol and ISDN showed dose-dependent relaxant effects on leukotriene (LT) D4-induced contraction of tracheal smooth muscle, though propranolol had no effect. 8-Bromo-cyclic GMP also showed a relaxant effect dose dependently. Nipradilol and ISDN elevated cyclic GMP levels in tracheal tissue dose dependently; however, propranolol caused no change in cyclic GMP levels. From these results, it is suggested that nipradilol relaxes LTD4-induced contraction of tracheal smooth muscle by increasing the tissue level of cyclic GMP.

Topics: Adrenergic beta-Antagonists; Animals; Cyclic GMP; Dose-Response Relationship, Drug; Guinea Pigs; In Vitro Techniques; Isosorbide Dinitrate; Leukotriene B4; Male; Muscle Contraction; Muscle Relaxation; Muscle, Smooth; Propanolamines; Propranolol; Trachea