cyclic-gmp and phenylhydrazine

Paroxysmal nocturnal hemoglobinuria (PNH) patients display exaggerated intravascular hemolysis and esophageal disorders. Since excess hemoglobin in the plasma causes reduced nitric oxide (NO) bioavailability and oxidative stress, we hypothesized that esophageal contraction may be impaired by intravascular hemolysis. This study aimed to analyze the alterations of the esophagus contractile mechanisms in a murine model of exaggerated intravascular hemolysis induced by phenylhydrazine (PHZ). For comparative purposes, sickle cell disease (SCD) mice were also studied, a less severe intravascular hemolysis model. Esophagus rings were dissected free and placed in organ baths. Plasma hemoglobin was higher in PHZ compared with SCD mice, as expected. The contractile responses produced by carbachol (CCh), KCl, and electrical-field stimulation (EFS) were superior in PHZ esophagi compared with control but remained unchanged in SCD mice. Preincubation with the NO-independent soluble guanylate cyclase stimulator 3-(4-amino-5-cyclopropylpyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine (BAY 41-2272; 1

Topics: Anemia, Sickle Cell; Animals; Cyclic GMP; Esophageal Diseases; Esophagus; Guanylate Cyclase; Hemolysis; Male; Mice; Mice, Inbred C57BL; Models, Animal; Muscle Contraction; Muscle, Smooth; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase Type III; Nitroprusside; Oxidative Stress; Phenylhydrazines; Pyrazoles; Pyridines; Signal Transduction; Soluble Guanylyl Cyclase

The same factors that regulate the activation of purified hepatic soluble guanylate cyclase by diverse agents possessing distinct requirements for enzyme activation were found to modulate cyclic GMP formation in intact viable hepatic cells. A comparison was made between activation of heme-deficient or heme-reconstituted guanylate cyclase and stimulation of cyclic GMP formation in mouse hepatic slices that were 95% viable and showed no active efflux of cyclic GMP. Heme-dependent activators of guanylate cyclase elicited a greater -fold increase in hepatic cyclic GMP levels in slices from phenobarbital-pretreated than control mice. Brilliant cresyl blue and KCN inhibited both enzyme activation and hepatic cyclic GMP accumulation caused by agents that generate nitric oxide. Hepatic slices from 3,5-diethoxycarbonyl-1,4-dihydrocollidine-treated mice, which are known to develop sharp increases in hepatic protoporphyrin IX/heme concentration ratios, showed elevated resting cyclic GMP levels whereas phenobarbital pretreatment produced decreased resting cyclic GMP levels compared to controls. Guanylate cyclase activation by azide required added catalase, and both enzyme activation and hepatic cyclic GMP formation were inhibited by aminotriazole. Enzyme activation by glyceryl trinitrate and NaNO2 required added thiols. Hepatic slices from acetaminophen-pretreated mice showed marked depletion of sulfhydryls and decreased cyclic GMP formation in response to these enzyme activators. Both effects were completely restored by treatment of thiol-depleted mice with N-acetylcysteine. These observations lend support to the general view that information gained from studies on the regulatory properties of purified soluble guanylate cyclase bears a close relationship to studies on regulatory mechanisms that modulate cyclic GMP formation in intact cells.

Topics: 1-Methyl-3-isobutylxanthine; Acetaminophen; Acetylcysteine; Animals; Cyclic AMP; Cyclic GMP; Guanylate Cyclase; Heme; Liver; Methylnitronitrosoguanidine; Mice; Nitroglycerin; Penicillamine; Phenobarbital; Phenylhydrazines; Potassium Cyanide; Probenecid; Protoporphyrins; Purinones; S-Nitroso-N-Acetylpenicillamine

The studies were carried out on male Wistar rats. The animals were divided into two groups: control and experimental. The hematocrit value and reticulocyte count were determined in blood samples collected from control and experimental rats. The experimental rats received subcutaneous injections of phenylhydrazine manufactured by Sigma for the duration of 3 days. On the 5th day blood samples were collected from all the control and experimental animals and determinations of hematocrit and reticulocyte count were repeated. cAMP levels were determined in bone marrow extracts by means of a radiocompetitive method. The cGMP level was determined by a radioimmunological assay. A significant elevation of cAMP level was detected in experimental rats, whereas the cGMP level changed only slightly.

Topics: Anemia; Animals; Bone Marrow; Cyclic AMP; Cyclic GMP; Hematocrit; Male; Phenylhydrazines; Rats; Rats, Inbred Strains; Reticulocytes