ABSTRACT
1. The antiarthritic and anti-inflammatory efficacy of N-acetyl-L-cysteine (NAC) was tested in male DBA/1 hybrid mice suffering from type II collagen-induced arthritis. Parameters including the arthritis index and the phagocytic responses recorded by chemiluminescence in unseparated blood were used for the assessment of disease activity. 2. Mice were immunized by subdermal injection of bovine type II collagen in Freund's complete adjuvant. The treatment with NAC started at day 42 after immunization and was continued over a period of six weeks: in doses ranging up to 50 mg/kg, a dose-dependent suppression of arthritis was noted; between 50 and 200 mg/kg, the inhibition curve had a plateau [ED50 = 50 mg/(kg x day)]. 3. The arthritis index correlated positively with the generation of chemiluminescence by reactive oxygen species (ROS) produced in neutrophils and monocytes activated by 12-O-tetradecanoylphorbol 13-acetate. 4. After treatment with 100 mg/kg of NAC from day 42 after immunization over a period of six weeks, the ROS production was reduced to levels occurring in whole blood of healthy animals. 5. It is concluded that low-molecular-weight antioxidants such as NAC may be adequate for controlling oxidative stress-derived damage in rheumatic diseases by modulation of ROS-dependent signal transduction pathways.
Subject(s)
Acetylcysteine/therapeutic use , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Arthritis/drug therapy , Collagen , Animals , Arthritis/chemically induced , Dose-Response Relationship, Drug , Luminescent Measurements , Male , Mice , Mice, Inbred DBAABSTRACT
Spermatozoa of 103 ejaculates from infertile patients and fertile healthy individuals were separated from seminal plasma and purified on Percoll gradient to determine the activities of superoxide dismutase (SOD) and catalase (CAT) in seminal plasma as well as in spermatozoal supernatants after hypotonic disintegration of the sperm plasma membrane. Out of collected specimens, a subgroup of ejaculates from 40 individuals was examined whose female partners had developed malignant processes in the cervix uteri (oncological subgroup). All sperm samples were classified into normal and pathological semen samples according to WHO criteria. While no significant differences of SOD levels were detected in seminal plasma of patients with primary infertility, a catalase deficiency seemed to be associated with combined sperm pathology-oligoasthenoteratozoospermia (OAT). Liberated concentrations of both SOD and catalase were diminished by 10-70% in the oncological subgroup compared to normozoospermia. In four OAT samples obtained from infertile males of the oncological subgroup, total depletion from both antioxidases was observed. A lack of sufficient antioxidase protection in cases of severe sperm pathology (OAT) may also lead to cervical dysplasia.
Subject(s)
Catalase/metabolism , Semen/enzymology , Spermatozoa/physiology , Superoxide Dismutase/metabolism , Female , Humans , Luminescent Measurements , Male , Spermatozoa/enzymologyABSTRACT
A range of compounds with a role in oxidative stress were measured in ejaculates from 40 normozoospermic individuals and 93 infertile males. Ejaculates were classified according to WHO criteria. Seminal plasma and the sperm cell fraction were assessed separately for superoxide dismutase (SOD), catalase, xanthine oxidase, capability for singlet oxygen trapping and content of thiobarbituric acid-reactive substances (TBARS). Pathological cases defined as oligozoospermia, asthenozoospermia, or teratozoospermia revealed different backgrounds of oxidative stress as reflected by different levels of tested substances in every type of sperm pathology. In the majority of abnormal ejaculates, a significant increase in intracellular activity of SOD, decreased intracellular levels of catalase, elevated levels of xanthine oxidase and TBARS, and severely impaired singlet oxygen trapping were observed when compared to normozoospermic ejaculates. Interrelationships between SOD and TBARS, and between xanthine oxidase and catalase, appeared to be of key importance when analysed separately in seminal plasma and in spermatozoa or in a combination of both. Elevated xanthine oxidase levels and low capacity for singlet oxygen trapping are statistically significant factors for the evaluation of male infertility which can develop as a result of persistent oxidative stress.
Subject(s)
Antioxidants/metabolism , Infertility, Male/metabolism , Reactive Oxygen Species , Semen/metabolism , Catalase/metabolism , Humans , Infertility, Male/enzymology , Male , Semen/enzymology , Spermatozoa/enzymology , Spermatozoa/metabolism , Superoxide Dismutase/metabolism , Thiobarbituric Acid Reactive Substances/metabolism , Xanthine Oxidase/metabolismABSTRACT
Oxidative damage caused by oxygen free radicals from activated phagocytes contributes to the pathology of arthritis. The present study evaluates the activity of NADPH oxidase of neutrophils and monocytes from patients suffering from various inflammatory and autoimmune rheumatic disease. Production rates of reactive oxygen species [ROS] of neutrophils and monocytes from rheumatic patients are compared to those of healthy controls and non rheumatic disease controls and correlated with the plasma levels of tumor necrosis factor alpha, C-reactive protein and the sedimentation rates of erythrocytes. There was a two- to eightfold increase in phagocytic superoxide production in rheumatic patients, when compared to healthy subjects or patients with non-rheumatic internal diseases [p < 0.005]. The enhanced NADPH oxidase-dependent superoxide generation correlated well with elevated levels of tumor necrosis factor alpha [TNF-alpha] in plasma [p = 0.005], suggesting a causal relation. There was no correlation with the plasma levels of C-reactive protein and a weak though significant correlation with the sedimentation rates of erythrocytes [p = 0.043]. Removal of circulating TNF-alpha by dialysis of patients blood and inhibition of NADPH oxidase by prednisolone treatment normalized elevated ROS production to the levels of healthy controls and correlated with the clinical improvements. Our data support the hypothesis of a central role for TNF-alpha during the development of arthritis. The chemiluminescence assay described here may be useful as a convenient screen and as a potential follow up procedure for individual patients with rheumatic diseases.
Subject(s)
Autoimmune Diseases/blood , Connective Tissue Diseases/blood , Inflammation/blood , Monocytes/enzymology , NADPH Oxidases/blood , Neutrophils/enzymology , Reactive Oxygen Species/metabolism , Rheumatic Diseases/blood , Tumor Necrosis Factor-alpha/analysis , Acridines , Blood Sedimentation , C-Reactive Protein/analysis , Combined Modality Therapy , Female , Granulomatosis with Polyangiitis/blood , Granulomatosis with Polyangiitis/therapy , Humans , Luminescent Measurements , Male , NADPH Oxidases/metabolism , Oxidative Stress , Phagocytosis , Prednisolone/pharmacology , Prednisolone/therapeutic use , Renal Dialysis , Superoxides/metabolism , Tumor Necrosis Factor-alpha/physiologyABSTRACT
Mitochondrial dysfunction contributes to cell damage in a number of human diseases. One significant mechanism by which mitochondria damage cells is by producing reactive oxygen species from the respiratory chain. In this study we measured the production of reactive oxygen species by leukocyte mitochondria in blood from rheumatoid arthritis patients. To do this we used the chemiluminescence of lucigenin, which is accumulated by mitochondria within cells and reacts with superoxide to form a chemiluminescent product. By using specific inhibitors we could distinguish between the production of reactive oxygen species by mitochondria and by NADPH oxidase. There was a five-fold increase in mitochondrial reactive oxygen species production in whole blood and monocytes from patients with rheumatoid arthritis, when compared to healthy subjects or patients with non-rheumatic diseases. There was no increases in mitochondrial reactive oxygen species production by neutrophils from rheumatoid arthritis patients. The enhanced mitochondrial radical production in rheumatoid arthritis patients correlated significantly with increased levels of tumor necrosis factor alpha in plasma (p < 0.0001). As tumor necrosis factor alpha is known to increase mitochondrial reactive oxygen species production the elevated mitochondrial radical formation seen in rheumatoid arthritis patients may be due to activation of the mitochondrial radical production. These data suggest that elevated mitochondrial oxidative stress contributes to the pathology of rheumatoid arthritis.
Subject(s)
Arthritis, Rheumatoid/blood , Leukocytes/ultrastructure , Mitochondria/metabolism , Reactive Oxygen Species/metabolism , Tumor Necrosis Factor-alpha/metabolism , Acridines , Antimycin A/pharmacology , Free Radicals , Humans , Luminescent Measurements , Monocytes/metabolism , NADPH Oxidases/metabolism , Neutrophils/metabolism , Oligomycins/pharmacology , Oxidative Stress , Potassium Cyanide/pharmacology , Rotenone/pharmacology , Uncoupling Agents/pharmacologyABSTRACT
We have studied the activity of substances (superoxide dismutase [SOD], catalase [Cat], malonaldehyde, xanthine oxidase [XO], nitric oxide [NOx]) participating in oxidative stress. Seminal plasma samples of 147 ejaculates obtained from normal and from infertile males were examined. Activities of SOD, Cat, and XO were measured chemiluminometrically while malonaldehydes and NOx were measured by spectrophotometer in seminal plasma samples. Ejaculates were previously characterized according to World Health Organization andrological criteria (sperm number, motility, and morphology). Procedures were performed in a university laboratory. Statistically significant changes (in comparison to normozoospermic samples) were noted in activities of SOD, XO, and malonaldehyde levels. The SOD activity exceeded values obtained for normozoospermic samples only in oligozoospermia. Otherwise low SOD levels in analyzed infertile subgroups inversely related to elevated malonaldehydes. Because diminished activity of SOD in seminal plasma was associated with increased levels of malonaldehydes and XO, we could postulate some significance of these monitored substances in evaluation of the cause of male infertility.
Subject(s)
Infertility, Male/physiopathology , Oxidative Stress/physiology , Catalase/metabolism , Humans , Male , Malondialdehyde/analysis , Nitric Oxide/analysis , Semen/chemistry , Semen/enzymology , Superoxide Dismutase/metabolism , Xanthine Oxidase/metabolismABSTRACT
Seminal plasma from ejaculates of 10 healthy, fertile volunteers and 63 infertile males was analysed for superoxide dismutase (SOD) and xanthine oxidase (XO) activities using a chemiluminometer. There was not statistically significant difference in the activity of either enzyme between control and infertile populations (113 +/- 74 IU/ml for SOD and 1.17 +/- 0.52 IU/ml for XO) in samples from normozoospermic ejaculates. Sperm progressive motility was positively correlated with SOD activity in seminal plasma of corresponding ejaculates (P < 0.05) and negatively with XO activity (P < 0.001). An 'oxido-sensitive' index was defined as the SOD/XO ratio and was found to be inversely related to sperm progressive motility samples (P < 0.01). Analysing this index among all tested samples of semen including those with pathological spermiograms, as well as normospermic (N) samples we found statistically significant (elevated) differences in oligoasthenoteratospermia (OAT) in comparison with N (P <0.05); OAT samples were also significantly different from oligospermic (O) and oligoteratospermic (OT) samples (P < 0.05). This suggests that the 'oxido-sensitive' index of seminal plasma may be a simple diagnostic factor, useful in the determination of male infertility.
Subject(s)
Infertility, Male/diagnosis , Semen/enzymology , Superoxide Dismutase/analysis , Xanthine Oxidase/analysis , Clinical Enzyme Tests , Humans , Luminescent Measurements , Male , Predictive Value of Tests , Reference Values , Sperm MotilityABSTRACT
The present study investigates synergistic effects of the TNF-alpha inhibitor thalidomide and the poly(ADP-ribose) polymerase (PARP)-inhibitor nicotinic acid amide (NAA) in male DBA/1 hybird mice suffering from type II collagen-induced arthritis. Parameters including the arthritis index, chemiluminescence and anti-collagen antibody titers were used for the assessment of disease activity: The disease courses demonstrated clearly an inhibitory effect of thalidomide. NAA inhibited established collagen arthritis in a dose-dependent manner. The combined application of thalidomide and NAA caused a powerful synergistic inhibition of arthritis. Furthermore, thalidomide and NAA were tested ex vivo for their inhibition of the NADPH oxidase-dependent generation of reactive oxygen species by activated neutrophils and monocytes in unseparated human blood. Our data show that type II collagen-induced arthritis can be suppressed by the simultaneous inhibition of TNF-alpha, PARP, and NADPH oxidase.
Subject(s)
Arthritis/drug therapy , Collagen/toxicity , Enzyme Inhibitors/therapeutic use , Niacinamide/pharmacology , Poly(ADP-ribose) Polymerase Inhibitors , Thalidomide/therapeutic use , Animals , Arthritis/chemically induced , Arthritis/enzymology , Arthritis/immunology , Arthritis, Rheumatoid , Cattle , Collagen/immunology , Disease Models, Animal , Drug Synergism , Enzyme Inhibitors/pharmacology , Humans , Immunization , Isoantibodies/blood , Male , Mice , Mice, Inbred DBA , NADPH Oxidases/antagonists & inhibitors , Neutrophils/drug effects , Respiratory Burst/drug effects , Thalidomide/pharmacology , Tumor Necrosis Factor-alpha/antagonists & inhibitorsABSTRACT
The effect of the parkinsonian neurotoxin, 1-methyl-4-phenylpyridinium (MPP+) together with nitric oxide donors on mitochondrial calcium homeostasis and membrane potential was investigated. Simultaneous exposure of calcium-loaded mitochondria to MPP+ and nitric oxide donors led to Cyclosporin A-sensitive mitochondrial calcium efflux and depolarisation. When MPP+ was replaced with the respiratory inhibitor rotenone, mitochondrial calcium efflux and depolarisation also occurred. As both MPP+ and rotenone induce mitochondrial superoxide formation, the possibility that calcium efflux and depolarisation were due to peroxynitrite formation from reaction of superoxide with nitric oxide was investigated. It was shown that simultaneous exposure of mitochondrial membranes to nitric oxide donors and rotenone led to peroxynitrite formation. The possible roles of nitric oxide, peroxynitrite, mitochondrial depolarisation, and calcium efflux in MPP+ toxicity are discussed.
Subject(s)
1-Methyl-4-phenylpyridinium/toxicity , Calcium/metabolism , Cyclosporine/pharmacology , Dopamine Agents/toxicity , Mitochondria, Liver/drug effects , Nitric Oxide/toxicity , Animals , Cattle , Drug Synergism , Female , Intracellular Membranes/drug effects , Membrane Potentials , Mitochondria, Heart/drug effects , Mitochondria, Liver/metabolism , Nitrates/metabolism , Rats , Rats, Wistar , Superoxides/metabolismABSTRACT
TH1-type proinflammatory cytokines induce the expression of phagocytic nitric oxide synthase (NOS) and prime the membrane-bound NADPH oxidase of neutrophils and monocytes of mice so as to attain an activated state, which upon a second stimulus releases up to 6-fold increased levels of reactive oxygen species (ROS) than do unprimed phagocytes. Enhanced levels of ROS and NO deregulate inflammatory signal transduction pathways, which play a crucial role in the pathogenesis of arthritis. The antiarthritic reactivity of diphenylene iodoniumchloride (DPI), an irreversible inhibitor of NADPH oxidase and NOS, was tested in male DBA/1xB10A(4R) hybrid mice suffering from potassium peroxochromate-induced arthritis. Daily doses of 2.8 mu mol/kg of DPI sufficed to inhibit the arthritis by 50%. A complete inhibition was obtained with 10 mu mol/kg of DPI. The reduction of overt arthritic symptoms correlated well with both the reduced levels of ROS and NO in plasma of DPI-treated mice. Our data support the hypothesis that oxidative stress and nitric oxides play a pivotal role in the pathology of arthritis, which can be therapeutically targetted by NADPH oxidase- and NO synthase-inhibitors.
Subject(s)
Arthritis/metabolism , NADPH Oxidases/antagonists & inhibitors , Nitric Oxide Synthase/antagonists & inhibitors , Animals , Anti-Inflammatory Agents , Arthritis/drug therapy , Arthritis/physiopathology , Chromates/pharmacology , Enzyme Inhibitors/pharmacology , Glutathione Reductase/antagonists & inhibitors , Inflammation/metabolism , Luminescent Measurements , Male , Mice , Mice, Inbred Strains , Nitric Oxide/metabolism , Nitric Oxide/pharmacology , Nitrites/metabolism , Onium Compounds/pharmacology , Peroxides/pharmacology , Reactive Oxygen Species/metabolism , Superoxides/metabolism , Tetradecanoylphorbol Acetate/pharmacologyABSTRACT
Recently, we demonstrated the hepatoprotective effects of nicotinic acid amide, a selective inhibitor of poly(ADP-ribose) polymerase (PARP; EC 2.4.2.30) on mice suffering from acetaminophen (AAP)-hepatitis, suggesting that the AAP-induced liver injury involves a step which depends on adenoribosylation. The present study investigates the effects of a diet free of precursors of NAD, the substrate on which PARP acts, in female NMRI mice with AAP hepatitis and evaluates the influence of simultaneous ethanol consumption in these animals. Liver injuries were quantified as serum activities of glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT). While AAP caused a 117-fold elevation of serum transaminase activities in mice kept on a standard laboratory diet, which was significantly exacerbated by ethanol and inhibited by nicotinic acid amide (NAA), adverse effects were noted in animals fed a diet free of precursors of NAD. In these animals, only minor increases of serum transaminase activities were measured in the presence of AAP, and unlike the exacerbation caused by ethanol in mice on a standard diet, the liver damage was inhibited by 50% by ethanol. A further 64% reduction of hepatitis was observed, when NAA was given to ethanol/AAP-mice. Our results provide evidence that the AAP-induced hepatitis and its exacerbation by ethanol can either be reduced by end-product inhibition of PARP by NAA or by dietary depletion of the enzyme's substrate NAD. We see the main application of NAA as for the combinational use in pharmaceutical preparations of acetaminophen in order to avoid hepatic damage in patients treated with this widely used analgesic.
Subject(s)
Acetaminophen/toxicity , Analgesics, Non-Narcotic/toxicity , Chemical and Drug Induced Liver Injury/prevention & control , Poly(ADP-ribose) Polymerases/pharmacology , Alanine Transaminase/blood , Alanine Transaminase/metabolism , Animals , Aspartate Aminotransferases/blood , Aspartate Aminotransferases/metabolism , Chemical and Drug Induced Liver Injury/enzymology , Chemical and Drug Induced Liver Injury/etiology , Ethanol/pharmacology , Female , Mice , Niacin/administration & dosage , Niacin/pharmacology , Tryptophan/administration & dosage , Tryptophan/pharmacologyABSTRACT
An array of therapeutically used analgetic and antirheumatic drugs causes severe liver damage. The present study investigates the hepatoprotective effects of inhibitors of NAD-dependent adenoribosylation reactions in analgesics-induced hepatic injury. Male NMRI mice were treated perorally with 500 mg/kg of acetaminophen, and the activities of both glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) were determined in serum. In addition, the activity of poly(ADP-ribose)polymerase (PARP) was quantified in liver cell nuclei. While the PARP-activity remained essentially unchanged, the acetaminophen-induced release of both GOT and GPT from injured liver cells could be inhibited by 90-99%, when mice were injected additionally with the selective PARP-inhibitors nicotinic acid amide, benzamide, caffeine, theophyline, and thymidine, respectively. We see the main application of inhibitors of adenoribosylation reactions as for the combinational use in pharmaceutical preparations of analgesics and antirheumatic drugs in order to avoid hepatic damage.
Subject(s)
Acetaminophen/toxicity , Alanine Transaminase/metabolism , Aspartate Aminotransferases/metabolism , Chemical and Drug Induced Liver Injury/prevention & control , Poly(ADP-ribose) Polymerases/pharmacology , Acetaminophen/antagonists & inhibitors , Alanine Transaminase/blood , Analgesics, Non-Narcotic/antagonists & inhibitors , Analgesics, Non-Narcotic/toxicity , Animals , Aspartate Aminotransferases/blood , Chemical and Drug Induced Liver Injury/etiology , Chemical and Drug Induced Liver Injury/metabolism , Male , MiceABSTRACT
1. The effects of racemic thalidomide (D[+]/L[-] alpha-phthalimido-glutarimide) on acetaminophen (AAP)-induced hepatitis were tested in male NMRI mice (n = 133) and quantified as serum activities of glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT). 2. A 2.1-fold increase of GOT and a 1.9-fold increase of GPT activities (P < 0.001) were observed in mice treated perorally with 500 mg/kg of AAP plus 150 mg/kg of thalidomide (Thal). In the absence of AAP, Thal did not display any detectable hepatotoxic effects. 3. The Thal-induced exacerbation of AAP hepatotoxicity was completely inhibited by nicotinic acid amide, a selective inhibitor of poly(ADP-ribose) polymerase (PARP) (P < 0.0001), suggesting a possible influence of Thal on the hepatic metabolism of NAD-adenoribosylation. 4. We see the main application of nicotinic acid amide as for the combinational use in pharmaceutical preparations of AAP in order to avoid hepatic damage in patients treated with AAP and Thal.
Subject(s)
Acetaminophen/adverse effects , Liver/drug effects , Niacinamide/pharmacology , Thalidomide/pharmacology , Alanine Transaminase/drug effects , Animals , Aspartate Aminotransferases/drug effects , Drug Interactions , Male , Mice , Mice, Inbred StrainsABSTRACT
1. The anti-inflammatory and hepatoprotective efficacy of CuPu(Py)2 ((N,N'-bis(2-pyridyl-methylene)-1,4-butanediamine) (N,N',N",N")-Cu2+), a serum-stable, copper-di-Schiffbase active centre analogue of Cu2Zn2 superoxide dismutase was tested in male NMNR mice suffering from endotoxin/galactosamine-induced hepatitis. 2. Parameters including the activities of serum transaminases and sorbitol-dehydrogenase as well as the levels of reactive oxygen and nitrogen intermediates which were used to quantify the disease activity. 3. A dose-dependent inhibition of hepatic enzyme release was noted in the presence of 0.1-10 mg/kg of CuPu(Py)2. 4. The release of transaminases from damaged liver cells was reduced by 68% and paralleled the reduction of serum levels of nitric oxides. 5. Elevated levels of reactive oxygen species were normalized to those healthy controls. 6. The copper-free apochelate Pu(Py)2, which is unable to dismutate superoxide, did not display any anti-inflammatory reactivity.
Subject(s)
Liver/drug effects , Superoxide Dismutase/pharmacology , Alanine Transaminase/drug effects , Animals , Antioxidants , Aspartate Aminotransferases/drug effects , Chemical and Drug Induced Liver Injury , Copper/pharmacology , Male , Mice , Mice, Inbred Strains , Nitric Oxide/bloodABSTRACT
Arthritis develops in DBA/1xB10A(4R) mice and Wistar rats upon intraplantar injection of potassium peroxochromate (K3CrO8), and is here quantified by whole blood chemiluminescence (CL) and 99mpertechnetate-imaging (99mTcO4-), and related to overt disease symptoms (the arthritis index). During the aqueous decay of K3CrO8 to chromate (VI), the chromium(V)-bound oxygen is released as superoxide, hydroxyl radicals, singlet oxygen and hydrogen peroxide, the same reactants, which are produced by activated phagocytes during inflammation. Reactive oxygen species (ROS) trigger the breakdown of the sulfhydryl-dependent antioxidant defence system and induce the nuclear factor kappa B-dependent expression of pro-inflammatory cytokines, which prime phagocytic NADPH oxidases to the enhanced production of ROS. During both the acute inflammatory response and the onset of the secondary response in non-injected paws, the phorbolester-stimulated ROS production of phagocytes was significantly enhanced (p < 0.001) and correlated well to the arthritis index (r = 0.797) and the uptake of 99mTcO4- into inflamed joints. Chromate(VI), formed during the decay of K3CrO8, contributes to the progression of arthritis by inhibition of glutathione reductase, thereby increasing intracellular H2O2 concentrations. In addition, Cr(VI) reduced to Cr(V) by ascorbate, catalyzes hydroxyl radical production in the presence of hydrogen peroxide. A stable loop forms, in which ROS, continuously produced by Cr(VI)/Cr(V) redox-cycling, drive the primary response into chronic self-perpetuating inflammation. We see the main application of K3CrO8-induced arthritis and its assessment by both 99mTcO4- imaging and chemiluminescent immunosensoring of phagocytic activity in unseparated blood as for the rapid screening of novel anti-rheumatic drugs and treatments.
Subject(s)
Arthritis/chemically induced , Chromates , Peroxides , Animals , Arthritis/blood , Arthritis/diagnostic imaging , Disease Models, Animal , Luminescent Measurements , Male , Mice , Mice, Inbred DBA , Phagocytosis , Radionuclide Imaging , Rats , Rats, Wistar , Sodium Pertechnetate Tc 99mABSTRACT
OBJECTIVE: The anti-arthritic reactivity of the IL-1 receptor antagonist IL-1ra was tested in male DBA/1xB10A(4R) mice suffering from potassium peroxochromate-induced arthritis. METHODS: Inflammatory arthritis was induced in male DBA/1xB10A(4R) mice by the intraplantar application of potassium peroxochromate and was quantified by the determination of the arthritis index, whole blood chemiluminescence, serum sulfhydryls and nitric oxides. Culture supernatants of the fibroblasts were assayed spectrophotometrically for collagenase release. RESULTS: Overt arthritic symptoms, as measured by the arthritis index, were significantly inhibited after two intraperitoneal injections of 2 mg/kg IL-1ra. The phorbol-ester-stimulated chemiluminescent response of monocytes and polymorphonuclear leukocytes in whole murine blood, which increased 5-fold during the development of arthritis, was normalized to the level of healthy controls. The oxidation of serum sulfhydryls was inhibited by 25%, and the serum levels of nitric oxides was depressed by 40%. In vitro the IL-1-dependent induction of collagenase of rabbit synovial fibroblasts was completely suppressed in the presence of 0.4 mg/ml IL-1ra. CONCLUSION: IL-1ra may prove useful to control the consequences of enhanced free radical production in inflammatory rheumatic diseases.
Subject(s)
Arthritis/drug therapy , Collagenases/metabolism , Fibroblasts/drug effects , Interleukin-1/metabolism , Sialoglycoproteins/pharmacology , Synovial Membrane/drug effects , Animals , Arthritis/blood , Arthritis/chemically induced , Arthritis/enzymology , Chromates , Fibroblasts/enzymology , Interleukin 1 Receptor Antagonist Protein , Luminescent Measurements , Male , Mice , Mice, Inbred DBA , Nitric Oxide/blood , Peroxides , Random Allocation , Recombinant Proteins/pharmacology , Synovial Membrane/cytology , Synovial Membrane/enzymology , Time FactorsABSTRACT
Proinflammatory cytokines prime the membrane-bound NADPH oxidase of neutrophils and monocytes of mice suffering from experimental arthritis so as to attain an activated state, which, upon a second stimulus, releases 6-fold increased levels of reactive oxygen species (ROS) than do unprimed phagocytes. Enhanced NADPH oxidase activity deregulates ROS-dependent signal transduction pathways of inflammation, which play a crucial role in the pathogenesis of arthritis. The antiarthritic reactivity of two inhibitors of NADPH oxidase, diphenylene iodoniumchloride (DPI) and staurosporine, was tested in male DBA/1 x B10A(4R) hybrid mice suffering from potassium peroxochromate arthritis. Daily doses of 2.8 mumol/kg of DPI or 30 nmol/kg of staurosporine sufficed to inhibit the arthritis by 50%. A complete inhibition was obtained with 10 mumol/kg of DPI, and 100 nmol/kg of staurosporine suppressed the arthritis by 85%. The onset, progression, and remission of arthritis correlated to both the activity of phagocytic NADPH oxidase (r = 0.750) and to overt disease symptoms as judged by the arthritis index. Our data support the hypothesis that oxidative stress plays a pivotal role in the pathology of arthritis, which can be therapeutically targeted by NADPH oxidase inhibitors.
Subject(s)
Alkaloids/therapeutic use , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Arthritis/prevention & control , Biphenyl Compounds/therapeutic use , NADH, NADPH Oxidoreductases/antagonists & inhibitors , Onium Compounds/therapeutic use , Oxidative Stress/drug effects , Phagocytes/drug effects , Reactive Oxygen Species/metabolism , Respiratory Burst/drug effects , Alkaloids/pharmacology , Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Arthritis/chemically induced , Biphenyl Compounds/pharmacology , Chromates/toxicity , Luminescent Measurements , Male , Mice , Mice, Inbred DBA , NADPH Oxidases , Onium Compounds/pharmacology , Peroxides/toxicity , Phagocytes/enzymology , Specific Pathogen-Free Organisms , StaurosporineABSTRACT
Poly(ADPR) polymerase (PARP; EC 2.4.2.30) is a nuclear enzyme, which, when activated by oxygen- and nitrogen-radical-induced DNA strand breaks, transfers ADP ribose units to nuclear proteins and initiates apoptosis by depletion of cellular NAD and ATP pools. The present study investigates whether the oxidative stress-dependent activation of PARP plays a role in the etiopathogenesis of arthritis. The antiarthritic reactivity of the biogenic PARP inhibitor nicotinamide was tested in DBA/1 x B10A(4R) mice suffering from potassium peroxochromate-induced arthritis. Daily doses of 4 mmol/kg of NA suppressed the arthritis by 35% and inhibited the phagocytic generation of reactive oxygen species, which increases sixfold during the development of arthritis. The onset, progression, and remission of arthritis correlated positively to the phorbolester-activated respiratory burst of neutrophils and monocytes, and a dose-dependent inhibition of NADPH oxidase activity was determined with human phagocytes. Our data support the hypothesis that oxidative stress-induced alterations in cellular signal transduction pathways play a pivotal role in the development of arthritis, which can be suppressed by the simultaneous inhibition of poly(ADPR) polymerase and NADPH oxidase.
Subject(s)
Arthritis/drug therapy , Niacinamide/pharmacology , Poly(ADP-ribose) Polymerase Inhibitors , Animals , Apoptosis , Arthritis/chemically induced , Arthritis/enzymology , Chromates/toxicity , Enzyme Activation/drug effects , Humans , Luminescent Measurements , Male , Mice , Mice, Inbred DBA , NADH, NADPH Oxidoreductases/antagonists & inhibitors , NADH, NADPH Oxidoreductases/metabolism , NADPH Oxidases , Oxidative Stress , Peroxides/toxicity , Poly(ADP-ribose) Polymerases/physiology , Reactive Oxygen Species/metabolism , Respiratory Burst/drug effects , Signal Transduction/drug effects , Tetradecanoylphorbol Acetate/pharmacologyABSTRACT
Reported is the case of a 60-year-old female who was referred to our rheumatology center because of a high titer of antinuclear antibodies (ANAs). Initially several lupus-like features suggested a diagnosis of undifferentiated connective tissue disease (UCTD). Immunofluorescence of the patient's serum showed a nuclear dot pattern. Counterimmunoelectrophoresis for antibodies to Sm, RNP, SS-A and SS-B were negative. Immunoblotting revealed a band at 53 kD which did not correspond to known autoantigens. Several months later the patient presented to the emergency room with symptoms due to a hypertensive crisis. Check-up finally revealed a cortisone-producing tumour of the right adrenal gland. The adenoma was removed by surgery and the patient did well again.
Subject(s)
Adrenal Gland Neoplasms/diagnosis , Adrenal Gland Neoplasms/immunology , Antibodies, Antinuclear/analysis , Connective Tissue Diseases/diagnosis , Connective Tissue Diseases/immunology , Diagnosis, Differential , Female , Fluorescent Antibody Technique , Humans , Immunoblotting , Immunoelectrophoresis , Middle Aged , Tomography, X-Ray ComputedABSTRACT
The anti-retroviral activity of Cu2Zn2 superoxide dismutase (SOD; EC 1.15.1.1) was tested in Molt-4 cells infected with the human immunodeficiency virus type 1 (HIV-1) and compared to the anti-HIV-1 activity of the reverse transcriptase inhibitors azidothymidine (AZT), dideoxycytidine (ddC), dideoxyuridine (ddU) and phosphono-formic acid, the glucosidase I inhibitors castanospermine and dihydroxymethyl dihydroxy-pyrrolidine (DMDP), the HIV protease inhibitor RO-31-7595 as well as the CD4-masking compound aurintricarboxylic acid. 300 nM of SOD sufficed to reduce the release of the viral antigen gp 120 of HIV-1NDK-infected Molt-4 cells by 50% [EC50]. Cytotoxic effects of SOD were estimated by cell counts and rates of cell growth. SOD, 3 µM, reduced the cell growth of uninfected cells by 50% [TC50]. While copper-free apo-SOD displayed no anti-HIV activity, the [EC50] of heat-inactivated enzyme was 1 µM, suggesting an anti-retroviral effect of low molecular weight active center degradation products of SOD. The [EC50] of SOD reached 10% of AZT's anti-HIV-1NDK activity and exceeded all tested anti-retrovirals 40-3000-fold. The selectivity index (Si= [TC50]/[EC50) for SOD was 10, resembling the reverse transcriptase inhibitors dideoxycytidine and phosphonoformic acid. SOD inhibited also dose-dependently the oxidative stress induced depletion of sulfhydryls, which are crucially involved in the nuclear factor kappa B controlled HIV transcription.