(Circulation. 1997;95:489-496.)
© 1997 American Heart Association, Inc.
Articles |
Nitric Oxide Contributes to the Progression of Myocardial Damage in Experimental Autoimmune Myocarditis in Rats
the Second Department of Medicine (S.I., M.H., S.A., T.S., H.I., F.M.), Tokyo Medical and Dental University, Tokyo; Departments of Pathology (S.I., T.N., T.K.) and Kidney Surgery (S.O.), Tokyo Women's Medical College, Tokyo; Second Department of Medicine (K.Y., M.M.), Yamaguchi University, Yamaguchi; and Second Department of Physiology (M.U.M., H.N.), Tokai University, Tokai, Japan.
Correspondence to Michiaki Hiroe, MD, Cardiology Division, Second Department of Medicine, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113, Japan.
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Abstract |
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Background Excess amounts of NO produced by an inducible NO synthase (iNOS) in response to cytokines may be cytotoxic and can be destructive to tissue. We investigated the role of NO in the development of myocardial damage and the effects of aminoguanidine (AG), an inhibitor of iNOS, on experimental autoimmune myocarditis in rats.
Methods and Results Autoimmune myocarditis was induced in 20 Lewis rats by injection of porcine cardiac myosin. Ten of the 20 rats were administered AG. The severity of myocarditis was evaluated by measuring the size of myocarditic lesion and serum levels of CK-MB. Serum NO levels were determined using the Cd/Cu method. Tissue specimens were immunohistochemically examined for iNOS and nitrotyrosine. Histopathological study revealed extensive myocardial destruction and massive inflammatory cell infiltration in AG-untreated rats but only focal mononuclear cell infiltration in AG-treated rats. The mean percent areas of inflammatory lesions in the untreated and treated rats were 56±13% and 3±2%, respectively (P<.001). NO levels were 102±23 and 25±9 IU/L, respectively (P<.01). CK-MB levels were 68±13 and 16±13 nmol/L, respectively (P<.01). Superoxide production as measured with an ex vivo monitoring system was also significantly decreased in the treated rats. Nitrotyrosine relating to the generation of peroxynitrite was detected through immunostaining in the inflammatory lesions of untreated rats but not in those of treated rats.
Conclusions Excess amounts of NO produced by iNOS appear to contribute to the progression of myocardial damage in myocarditis. AG may prove to be useful in the treatment of myocarditis.
Key Words: free radicals • myocarditis • myosin • nitric oxide • inducible nitric oxide synthase
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Introduction |
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In humans, acute myocarditis is a potentially lethal disease. Although the precise mechanisms of the development of viral myocarditis are still unknown,1 2 its pathogenesis is the result of natural killer cells, viral-specific cytotoxic T cells,3 4 and autoantibodies.5 6 It has been reported that the pathogenesis of the tissue damage in viral myocarditis7 8 resembles that in experimental autoimmune myocarditis, which is induced by myosin. A relation has been demonstrated between acute CB3 murine or rat myocarditis and experimental autoimmune heart disease in several strains of mice9 10 or Lewis rat.11 Therefore, to understand the mechanisms of myocardial injury in myocarditis, we used an experimental model for autoimmune myocarditis induced in Lewis rats with the use of myosin as the autoantigen.8 12
NO has been found to be involved in diverse physiological and pathophysiological processes,13 including host immune defense, vasoregulation, neurotransmission,14 the pathogenesis of diabetes,15 and rejection of heart transplants.16 NOS, an enzyme that is involved in the synthesis of NO, has recently been shown to be activated in the inflammatory lesion. There are at least three distinct types of NOS: the constitutive types bNOS17 and eNOS18 and iNOS,19 which is induced de novo in macrophages, hepatocytes, mesangial cells, and endothelial cells by LPS and various cytokines (IL-1ß, TNF-
, IFN-
). The latter form leads to the production of large amounts of NO, and NO is thought to be in part responsible for the loss of vascular responsiveness seen in septic shock.20 During inflammation, NO is produced in large amounts by activated macrophages, neutrophils, and various other cells via a pathway catalyzed by iNOS. In addition to its role as a cellular messenger at low concentrations, NO is thought to be involved in pathological processes due to its cytotoxicity at high concentrations. iNOS can be expressed in diverse cell types and mediates, in part, the cytostatic and cytotoxic effector function of activated macrophages.21 22
AG, an inhibitor of iNOS,23 24 25 was recently demonstrated to decrease the severity of the pathophysiological sequelae of excess NO production in diabetes,15 26 uveitis,27 and experimental autoimmune encephalomyelitis.28
In the present study, we investigated the role of NO in the development of myocardial damage and the effects of AG on experimental autoimmune myocarditis in rats.
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Methods |
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Induction of Autoimmune Myocarditis and Experimental Designs
Autoimmune myocarditis was induced in rats as previously described.7 8 12 29 The protocol for these experiments was approved by the committee in charge of animal welfare of our hospital (Tokyo Medical and Dental University and Tokyo Women's Medical College). Four groups of female 7-week-old Lewis rats were used. Porcine heart myosin (Sigma Chemical Co) was dissolved in a 0.01 mol/L PBS at a concentration of 5 g/L. Twenty rats were injected with 0.1 mL (10 mg/kg) of myosin (1 g/L) mixed with an equal volume of Freund's complete adjuvant (Sigma) in a foot pad on days 1 and 7. On days 2 and 5, they were injected with 0.1 mL of Bordetella pertussis (Nacalai Tesque) dissolved in 0.9 mL of saline solution via the caudal vein. Ten of these animals were killed on day 21 (group A). The other 10 rats (group B) were injected with AG (Sigma) at a dosage of 400 mg/kg IP every day starting on the day before the first sensitization with porcine cardiac myosin until the day on which they were killed (day 21). Group C1 consisted of 5 control rats administered AG but not myosin, and group C2 consisted of 5 control rats injected with saline without AG. After the animals were killed, blood samples of
3 mL each were collected promptly from the heart to measure NO (NO2- + NO3-) and CK-MB. The hearts were then removed and divided to obtain specimens for histology, immunohistochemistry, and in situ hybridization.
Hemodynamic Measurements
Hemodynamic examination was performed after the administration of sodium pentobarbital (50 mg/kg IP) by a previously described method.30 In brief, on day 21, the jugular vein was cannulated with a polyethylene catheter connected to a Statham transducer (P10EZ, Gould). The catheter was advanced into the right ventricle. The external right carotid artery was exposed and cannulated with a micromanometer-tipped catheter (PR249, Miller Instruments). Measurements were thus obtained of both ventricular systolic and end-diastolic pressures, positive dP/dt of the left ventricle, aortic pressure, and heart rate through ECG monitoring.
Histopathological Study
Transverse sections of the heart specimens, which had been sliced into three equal portions, were prepared, fixed in phosphate-buffered 10% formalin, embedded in paraffin, cut into sections 3 µm thick, and stained with hematoxylin and eosin. The areas of the entire heart and of regions affected by myocarditis (ie, regions showing infiltration by inflammatory cells and myocardial necrosis) were determined with the use of a personal computer (Apple Computer EM software, Rise Co), and the area ratio (affected/entire area in percent) was calculated. Values for three ventricular portions were averaged for each heart. Mean area ratio (myocarditis-affected area ratio) in each group was then compared. These data were determined by two blind observers.
Immunohistochemical Study
Tissue specimens were fixed in periodate lysine-paraformaldehyde fixative and frozen in Optimal Cutting Temperature Compound (O.C.T. Compound) after a complete rinse with sucrose-containing PBS or fixed in phosphate-buffered 10% formalin. For immunostaining, frozen sections or deparaffinized sections were immersed in 3% hydrogen peroxidase in methanol for 30 minutes. Normal goat serum was then applied for 10 minutes. Sections were incubated with primary antibodies overnight at 4°C. The primary antibodies were OX19 (1:200 dilution, Serotec Inc), a marker for pan-T cells; W3/25 (CD4, 1:200, Serotec), a marker for helper T cells; OX8 (CD8, 1:200, Serotec), a marker for cytotoxic suppressor T cells; OX33 (1:200, Serotec), a marker for B cells; anti-iNOS antisera31 32 33 34 35 36 (1:300 dilution, Santa Cruz Biotechnology); anti-eNOS antisera (Calbiochem-Novablochem Co); and anti-nitrotyrosine antibody (1:500 dilution, Upstate Biotechnology). The PBS-washed sections were then incubated with goat anti-rabbit IgG-peroxidase conjugate (1:200) for 60 minutes at room temperature. Peroxidase activity was detected in Tris-buffered saline solution containing 3,3'-diaminobenzidine tetrahydrochloride and 0.1% H2O2 for iNOS and alkaline phosphate fast red for nitrotyrosine. The specimens were counterstained with hematoxylin. To assess staining specificity, control staining, including an immunoabsorption test, was performed simultaneously as previously described.37 38
Determination of serum NO (NO2- + NO3-)
Levels of NO (NO2- + NO3-) in serum were determined according to the Cd/Cu method using the TCI-NOX1000 system (Tokyo Kasei Kogyo Co).39 In principle, NO2- was reduced to NO3- with a high-performance Cd/Cu reduction column, and the total NO3- content was determined by high-performance liquid chromatography in an NO (NO2- + NO3-) assay system. The absorbance of this system was 546 nm.
Determination of serum CK-MB
After the rats were killed, serum was obtained from blood collected from the heart chamber. Levels of CK-MB, as a marker of cell injury in serum, were determined in a Type II Chemical Analyzer (Ciba Corning) with Chemilumi CK-MB Magic Lite (Ciba Corning) used as the reagent.
Determination of Superoxides
The ex vivo monitor system40 consisted of a light-proof box, a photomultiplier, and a photon counter, which were completely shielded from light. The hearts of the control animals and those in groups A, B, C1, and C2 were perfused with physiological saline buffer with O2 and were placed upside down in the black box. The O2-·-specific chemiluminescence probe MCLA, a Luciferin analogue, was introduced via a side arm. Chemiluminescence from the surface of the heart was counted every second (CPS) through a window (7-mm diameter) with a final concentration of MCLA of 55 mmol/L. The sensitivity of the system was as high as 4x10-16 mol of O2-· per second. The specificity of MCLA chemiluminescence for O2-· was examined with the use of isolated human leukocytes. The addition of phorbol-12-myristate-13-acetate led to an increase in photon counts in both probes.
In Situ Hybridization
A 604–base-pair fraction was excised from the noncoding region of murine macrophage iNOS cDNA, which was kindly provided by Drs C. Nathan and N. McCartney-Francis,41 to prepare an antisense probe and a sense probe (homology with rat, 94%) labeled with digoxigenin. Specimens of heart were fixed in 4% paraformaldehyde with 0.5% glutaraldehyde and embedded in paraffin with a low melting point. After deparaffinization in xylene, sections were digested with 100 g/L proteinase K in 10 mmol/L Tris-HCl, pH 7.6, and 1 mmol/L EDTA for 10 minutes at room temperature. The hybridization buffer contained 0.6 mol/L NaCl, 1 mmol/L EDTA, 10 mmol/L Tris-HCl, 10% dextran sulfate, 0.25% SDS, 200 g/L tRNA, 1x Denhardt's solution, 10 mmol/L dithiothreitol, and 50% deionized formamide. A volume of 50 µL of hybridization buffer that contained either the antisense or sense probe was applied to each section. This was followed by incubation in a moist chamber for 16 hours. Sections were washed in 2x SSC and 50% formamide for 30 minutes at 50°C, followed by incubation with 20 g/L RNase A (Boehringer-Mannheim) for 30 minutes at 37°C. After being washed in 2x SSC for 20 minutes and then washed in 0.2x SSC for 20 minutes twice at 42°C, immunological detection of the digoxigenin-labeled probe was accomplished using anti-digoxigenin-alkaline phosphatase. It was visualized with the use of 4-nitro blue tetrazolium chloride and 5-bromo-4-chloro-3-indolyl-phosphate.
Statistical Analysis
Results are expressed as mean±SD. Affected area (in percent), NO (NO2- + NO3-), CK-MB, and superoxides were compared by ANOVA. In hemodynamics, one-way ANOVA and a Sheffe's method of comparison analysis were used to determine the statistical significance of the differences among the four groups. A value of P<.05 was considered statistically significant.
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Results |
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Hemodynamics
In group A, significant weight loss was observed compared with groups B, C1, and C2. In group A, both left and right ventricular end-diastolic pressures were significantly increased, whereas the systolic left ventricular and mean aortic pressures were markedly decreased. A substantial decrease in positive dP/dt of the left ventricle was also evident in group A, indicating a global impairment of left ventricular function. Five parameters-left and right ventricular end-diastolic pressures, systolic left ventricular pressures, mean aortic pressures, and positive dP/dt of the left ventricle-in group B were similar to those in groups C1 and C2 (Table 1
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Histopathology
Tissue specimens from rats in group A exhibited marked infiltration by inflammatory cells, including lymphocytes, macrophages, neutrophils, and giant cells and extensive necrosis of cardiac muscle cells (Fig 1A). In contrast, those from rats in group B exhibited only focal infiltration by inflammatory cells, and necrosis was limited to cardiac myocytes that were in contact with the inflammatory cells (Fig 1B). No infiltration by inflammatory cells or myocyte necrosis was noted in either control group (Fig 1C and 1D, groups C1 and C2, respectively).
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Affected Area Ratio
The myocarditis-affected area ratios in groups A and B were 56±13% and 3±2%, respectively (Fig 2). The value in group B was significantly low compared with that in group A. Values in control groups (C1 and C2) were both 0% (Fig 2).
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Immunohistochemistry
In tissue specimens from rats in group A, macrophages, neutrophils, giant cells, vascular endothelial cells, and vascular smooth muscle cells stained positive for iNOS (Fig 3A). Infiltrating lymphocytes showed more predominantly immunopositivity for CD4 than for CD8 in the inflammatory lesion. Myocardial cells in the inflammatory lesion also showed positive immunostaining for iNOS (Fig 3A). An immunopositive substance was present diffusely in the cytoplasm of myocytes. Unlike immunostaining with the anti-iNOS antibody, immunohistochemical investigation with the anti-eNOS antibody revealed positivity only in vascular endothelial cells (data not shown). Preabsorption of the iNOS antiserum with iNOS antigen completely abolished immunostaining (data not shown). It was confirmed by these results that the anti-iNOS antibody showed no cross-reaction with the anti-eNOS antibody. Immunohistochemical staining with anti-nitrotyrosine antibody revealed positive reactivity in the damaged myocytes (Fig 3B).
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In tissue specimens from rats in groups B, some of the focally infiltrating inflammatory cells and a small number of cardiac muscle cells in contact with the inflammatory cells were iNOS positive (Fig 3C
), and a small number of cardiac muscle cells in contact with inflammatory cells were nitrotyrosine negative (Fig 3D). The two control groups (groups C1 and C2) exhibited no staining for iNOS (Fig 3E1 and 3E2) or nitrotyrosine (data not shown).
Determination of Serum NO (NO2- + NO3-)
NO (NO2- + NO3-) values in groups A (n=10), B (n=10), C1 (n=5), and C2 (n=5) were 102±23, 25±9,* 12±3,* and 10±10* nmol/mL, respectively (*P<.01 versus group A) (Fig 4). There was a significant difference between groups A and B and between groups A and C1 or C2.
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Determination of Serum CK-MB
CK-MB values in groups A (n=10), B (n=10), C1 (n=5), and C2 (n=5) were 68±13, 16±13,* 6±3,* and 9±6* IU/L, respectively (*P<.001 versus group A) (Fig 5). There was a significant difference between groups A and B and between groups A and C1 or C2.
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Superoxide Levels
Studies of chemiluminescence ex vivo revealed a clear difference among the three groups. The peak count was significantly higher in group A (1035±212 CPS) versus group B (188±103 CPS), group C1 (159±78 CPS), and group C2 (153±62 CPS) (P<.05) (Fig 6).
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In Situ Hybridization
In group A, the in situ hybridization study revealed positive reactivity for the iNOS mRNA antisense probe in the myocytes involved in the inflammatory lesion, as well as in the vascular smooth muscle cells, vascular endothelial cells, and macrophages (Fig 7A and 7B). The myocardium in groups C1 and C2 that was not involved in the inflammation in a heart showed negative reactivity (Fig 7C). The sense probe showed also negative reactivity (Fig 7D). In group B, the in situ hybridization study revealed focally positive reactivity for the iNOS mRNA antisense probe in the myocytes in contact with inflammatory cells and macrophages (data not shown).
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Discussion |
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The results of the present study demonstrate that iNOS was expressed in the myocardium and a large amount of NO and superoxide (O2-·) was produced in the heart of rats with experimental myocarditis. It was also demonstrated that myocytes stained positive for nitrotyrosine in the affected area. These findings indicate that the production of excess amounts of NO by iNOS reacts with superoxide and forms a peroxynitrite, changing tyrosine in myocardium to nitrotyrosine and leading to myocardial injury in the autoimmune myocarditis of rats.
There have been no previous reports of an increase of iNOS in myocarditis, although Belder et al42 described a significant increase in myocardial iNOS activity in dilated cardiomyopathy. Numerous reports have shown that iNOS is induced by LPS and by cytokines such as IFN-, TNF-, and IL-1ß.19 43 Schulz et al44 reported that iNOS is induced in isolated cardiac myocytes from guinea pig hearts exposed to IFN- and to bacterial endotoxin. Roberts et al45 demonstrated that IL-1ß induces iNOS activity and iNOS mRNA levels in cardiac myocytes from neonatal rats maintained in vitro. We reported the expression of human myocardial iNOS in acute myocarditis, suggesting that the enhanced production of NO by iNOS is cytotoxic and accounts, in part, for myocardial injury and reduced myocardial contractility during acute illness.46 Large amounts of cytokines are released from inflammatory cells in myocarditis, including macrophages and lymphocytes, in the myocardial tissue.47 Thus, it seems reasonable that iNOS would be induced in the tissue of a myocarditic heart. Using a specific antiserum against iNOS, whose specificity was confirmed by various immunoabsorption tests and other relevant examinations and which shows no cross-reaction with the anti-eNOS antibody, positive reactivity was detected in the cytoplasm of macrophages, neutrophils, vascular smooth muscle or endothelial cells, and cardiac myocytes in the inflammatory lesion. The cytokines released by the surrounding inflammatory cells were likely responsible for the induction of iNOS in these cells. iNOS mRNA was strongly expressed in the myocardial cells similar to the protein levels in the present study. The induction of iNOS in the myocardium caused an extensive and prolonged release of NO. NO (NO2- + NO3-) levels were significantly higher in the serum and heart of rats with myocarditis compared with control rats.
Superoxide (O2-·) is generated in the tissue with inflammation or ischemia/reperfusion.48 49 It was demonstrated in the present study that a large amount of O2-· was produced in the heart with myocarditis. Steuer et al50 suggested that NO caused cell injury by damaging iron and sulfur proteins, but attention has recently focused on other nitrogen oxides, including peroxynitrite. Peroxynitrite, formed from NO, is a powerful oxidant and causes tissue damage.51 Peroxynitrite formation can be estimated immunohistochemically using anti-nitrotyrosine antibody because nitrotyrosine is a major product of the attack of peroxynitrite on proteins.52 53 54 We observed positive immunoreactivity for nitrotyrosine in the damaged myocytes of rats with myocarditis. Peroxynitrite may also form hydroxyl radicals under certain conditions that, in turn, may damage tissues. It is suggested that NO is cytotoxic to the myocardial cells and that peroxinitrite may be a major cytotoxic agent in myocarditis.
The present study also demonstrated that clinical features and hemodynamics were significantly improved in the AG-treated group. In addition, it was demonstrated that histopathological findings of myocardial inflammation were significantly less pronounced, area ratios of inflammatory lesions were significantly smaller, and serum CK-MB and blood NO (NO2- + NO3-) levels were significantly lower in rats with myocarditis that had been administered AG than in those that had not been administered AG. In the former group, a small number of cardiac muscle cells in contact with inflammatory cells were iNOS positive but nitrotyrosine negative. These findings indicate that in rats with autoimmune myocarditis treated with AG, an iNOS inhibitor, iNOS is expressed but there is minimal production of NO and, therefore, inflammatory myocardial injury is significantly reduced. The number of infiltrating inflammatory cells was quite small in the AG-treated group. However, this finding could not be attributed to inhibition of inflammatory cell migration because there is no evidence that AG inhibits cytokines or directly inhibits the infiltration of inflammatory cells. Thus, impairment of the myocardium in myocarditis may be the result of the following cycle; an initial focal myocarditic inflammatory reaction would occur, and then further infiltration of inflammatory cells would be induced by the resultant myocardial cell destruction. These cells would release cytokines, extending the area of myocardial damage by various factors, including NO. It is possible that AG breaks this vicious cycle by inhibiting inflammation with reduction of NO production at the early stages.
In conclusion, iNOS was highly expressed in myocardial cells within the inflammatory lesion in experimental rat myocarditis. In turn, iNOS induced the production of excess amounts of NO, which reacts with superoxide, producing peroxynitrite, a highly cytotoxic compound. In addition, AG, an inhibitor of NOS, prevented the progression of myocardial lesions. Therefore, it is suggested that NO participated in the pathogenesis of myocardial injury in this model of autoimmune myocarditis rat. Further study should determine whether specific inhibition of iNOS will be of therapeutic benefit in this condition of vast overproduction of NO.
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Selected Abbreviations and Acronyms |
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Acknowledgments |
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This study was supported by a Grant-in Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan (06670746, 07670765); by a grant from the Japan Private School Promotion Foundation; by a grant from the Japan Research Promotion Society for Cardiovascular Diseases; and by a Research Grant for Diseases from the Ministry of Health and Welfare of Japan.
Received June 17, 1996; revision received August 14, 1996; accepted August 28, 1996.
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References |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Lieberman EB, Hutchins GM, Herskowitz A, Rose NR, Baughman KL. Clinicopathologic description of myocarditis. J Am Coll Cardiol.. 1991;18:1617-1626.[Abstract]
2.
Mason JW, O'Connell JB, Herskowitz A, Rose NR, McManus BM, Billingham ME, Moon TE. The myocarditis treatment trial investigators: a clinical trial of immunosuppressive therapy for myocarditis. N Engl J Med.. 1995;333:269-275.
3. Huber SA, Job LP, Woodruff JF. Lysis of infected myofibers by coxsackievirus B3-immune T lymphocytes. Am J Pathol.. 1980;98:681-694.[Abstract]
4. Alvarez FL, Neu N, Rose NR, Craig SW, Beisel KW. Heart-specific autoantibodies induced by coxsackievirus B3: identification of heart autoantigens. Clin Immunol Immunopathol.. 1987;43:129-139.[Medline] [Order article via Infotrieve]
5.
Wolfgram LJ, Beisel KW, Rose NR. Heart-specific autoantibodies following murine coxsackievirus B3 myocarditis. J Exp Med.. 1985;161:1112-1121.
6. Deguchi H, Hayashi Y, Kotaka M, Kawamura K. Cell-mediated immune cardiocyte injury in viral myocarditis of mice and patients. Jpn Circ J.. 1989;53:61-77.[Medline] [Order article via Infotrieve]
7. Pummerer C, Berger P, Fruhwirth M, Ofner C, Neu N. Cellular infiltrate, major histocompatibility antigen expression and immunopathogenic mechanisms in cardiac myosin-induced myocarditis. Lab Invest.. 1991;65:538-547.[Medline] [Order article via Infotrieve]
8. Neumann DA, Lane JR, Wulff SM, Allen GS, Lafond-Walker A, Herskowitz A, Rose NR. In vivo deposition of myosin-specific autoantibodies in the hearts of mice with experimental autoimmune myocarditis. J Immunol.. 1992;148:3806-3813.[Abstract]
9. Neu N, Rose NR, Beisel KW, Herskowitz A, Gurri-Glass G, Craig SW. Cardiac myosin induces myocarditis in genetically predisposed mice. J Immunol.. 1987;139:3630-3636.[Abstract]
10. Neu N, Craig SW, Beisel KW, Rose NR. Coxsackievirus induced autoimmune myocarditis in mice: cardiac myosin autoantibodies do not cross-react with the virus. Clin Exp Immunol.. 1987;69:566-574.[Medline] [Order article via Infotrieve]
11.
Kodama M, Matsumoto Y, Fujiwara M. In vivo lymphocyte-mediated myocardial injuries demonstrated by adoptive transfer of experimental autoimmune myocarditis. Circulation.. 1992;85:1918-1926.
12.
Kodama M, Matsumoto Y, Fujiwara M, Zhang S, Hanawa H, Itoh E, Tsuda T, Izumi T, Shibata A. Characteristics of giant cells and factors related to the formation of giant cells in myocarditis. Circ Res.. 1991;69:1042-1050.
13. Moncada S, Palmer RMJ, Higgs EA. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev.. 1991;43:109-142.[Medline] [Order article via Infotrieve]
14. Nathan C. Nitric oxide as a secretory product of mammalian cells. FASEB J.. 1992;6:3051-3064.[Abstract]
15. Corbett JA, Tilton RG, Chang K, Hasan KS, Ido Y, Wang JL, Sweetland MA, Lancaster JR, Jr, Williamson JR, McDaniel ML. Aminoguanidine, a novel inhibitor of nitric oxide formation, prevents diabetic vascular dysfunction. Diabetes.. 1992;41:552-556.[Abstract]
16.
Worrall NK, Lazenby WD, Misko TP, Lin T-S, Rodi CR, Manning PT, Tilton RG, Williamson JR, Ferguson TB, Jr. Modulation of in vivo alloreactivity by inhibitor of inducible nitric oxide synthase. J Exp Med.. 1995;181:63-70.
17. Bredt DS, Hwang PM, Glatt CE, Lowenstein C, Reed RR, Snyder SH. Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase. Nature.. 1991;351:714-718.[Medline] [Order article via Infotrieve]
18. Marsden AP, Schappert TK, Michel T. Molecular cloning and characterization of human endothelial nitric oxide synthase. FEBS Lett.. 1992;307:287-293.[Medline] [Order article via Infotrieve]
19. Nathan C, Xie Q-W. Nitric oxide synthases: roles, tolls, and controls. Cell.. 1994;78:915-918.[Medline] [Order article via Infotrieve]
20.
Baok KJ, Thiel BA, Stuehr DJ. Macrophage nitric oxide synthase subunits. J Biol Chem.. 1993;268:21120-21129.
21. Hibbs JB, Taintor RR, Jr, Vavrin Z, Granger DL, Drapier J-C, Amber IJ, Lancaster JR, Jr. Synthesis of nitric oxide from a terminal guanidino nitrogen atom of L-arginine: a molecular mechanism regulating cellular proliferation that targets intracellular iron. In: Moncada S, Higgs EA, eds. Nitric Oxide From L-Arginine: A Bioregulatory System. Amsterdam, Netherlands: Elsevier; 1990:189-223.
22.
Stuehr DJ, Nathan CF. Nitric oxide: a macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J Exp Med.. 1989;169:1543-1555.
23. Misko TP, Moore WM, Kasten TP, Nickols GA, Corbett JA, Tilton RG, McDaniel ML, Williamson JR, Currie MG. Selective inhibition of the inducible nitric oxide synthase by aminoguanidine. Eur J Pharmacol.. 1993;233:119-125.[Medline] [Order article via Infotrieve]
24.
Crbett JA, Mikhael A, Shimizu J, Frederick K, Misko TP, Kanagawa O, Unanue ER. Nitric oxide production in islets from nonobese diabetic mice: aminoguanidine-sensitive and resistant stages in the immunological diabetic process. Proc Natl Acad Sci U S A.. 1993;90:8992-8995.
25. Misko TP, Moore WM, Kasten TP, Nickols GA, Corbett JA, Tilton RG, McDaniel ML, Williamson JR, Currie MG. Selective inhibition of the inducible nitric oxide synthase by aminoguanidine. Eur J Pharmacol.. 1993;233:119-125.
26. Tilton RG, Chang K, Hasan KS, Smith SR, Petrash JM, Misko TP, Moore WM, Currie MG, Corbett JA, McDaniel ML, Williamson JR. Prevention of diabetic vascular dysfunction by guanidines: inhibition of nitric oxide synthase versus advanced glycation end-product formation. Diabetes.. 1993;42:221-232.[Abstract]
27.
Tilton RG, Chang K, Corbett JA, Misko TP, Currie MG, Bora NS, Kaplan HJ, Williamson JR. Endotoxin-induced uveitis in the rat is attenuated by inhibition of nitric oxide production. Invest Ophthalmol Visual Sci.. 1994;35:3278-3288.
28. Cross AH, Misko TP, Lin RF, Hickey WF, Trotter JL, Tilton RG. Aminoguanidine, an inhibitor of inducible nitric oxide synthase, ameliorates experimental autoimmune encephalomyelitis in SJL mice. J Clin Invest.. 1994;93:2684-2690.
29. Hanawa H, Tsuchida M, Matsumoto Y, Watanabe H, Abo T, Sekikawa H, Kodama M, Zhang S, Izumi T, Shibata A. Characterization of T cells infiltrating the heart in rats with experimental autoimmune myocarditis. J Immunol.. 1993;150:5682-5695.[Abstract]
30.
Hiroe M, Ohta Y, Fujita N, Nagata M, Toyozaki T, Kusakabe K, Sekiguchi M, Marumo F. Myocardial uptake of 111In monoclonal antimyosin Fab in detecting doxorubicin cardiotoxicity in rats. Circulation.. 1992;86:1965-1972.
31. Schmidt HHHW, Walter U. NO at work. Cell.. 1994;78:919-925.[Medline] [Order article via Infotrieve]
32.
Farias-Eisner R, Sherman MP, Aeberhard E, Chaudhuri G. Nitric oxide is important mediator for tumoricidal activity in vivo. Proc Natl Acad Sci U S A.. 1994;91:9407-9411.
33. Marietta MA. Nitric oxide synthase: aspects concerning structure and catalysis. Cell.. 1994;78:927-930.[Medline] [Order article via Infotrieve]
34.
Bukrinsky ME, Nottet HS, Schmidtmayerova N, Dubrovsky L, Flanagan CR, Mullins ME, Lipton SA, Gendelman HE. Regulation of nitric oxide synthase activity in human immunodeficiency virus type 1 (HIV-1)-infected monocytes: implications for HIV-associated neurological disease. J Exp Med.. 1995;181:735-745.
35. Heiss LN, Lancaster JRJr, Corbett JA, Goldman WE. Epithelial autotoxicity of nitric oxide: role in the respiratory cytopathology of pertussis. Proc Natl Acad Sci U S6A.. 1994;91:267-270.
36.
Kamijo R, Harada H, Matsuyama T, Bosland M, Gerecitano J, Shapiro D, Le J, Im KS, Kimura T, Green S, Mak TW, Taniguchi T, Vilcek J. Requirement for transcription factor IRF-1 in NO synthase induction in macrophages. Science.. 1994;263:1612-1615.
37. Nishikawa T, Kasajima T, Naruse M, Naruse K, Demura H, Hiroe M, Nakazawa M, Nakajima Y, Nagata M. Immunohistochemical study on human atrial natriuretic polypeptide in the ventricle of hearts with endocardial fibroelastosis. Am J Cardiovasc Pathol.. 1990;3:247-25138.[Medline] [Order article via Infotrieve]
38. Hiroe M, Hirata Y, Marumo F, Nagata M, Toyozaki T, Hasumi M, Ohta Y, Horie T, Sekiguchi M. Immunohistochemical localization of endothelin in human vascular endothelial cells. Peptides.. 1989;10:1281-1282.[Medline] [Order article via Infotrieve]
39. Green LC, Wagner DA, Glogowski J, Skipper P, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem.. 1982;126:131-138.[Medline] [Order article via Infotrieve]
40. Pronai L, Nakazawa H, Ichimori K, Saigusa Y, Ohkubo T, Hiramatsu K, Arimori S, Feher J. Time course of superoxide generation by leukocytes: the MCLA chemiluminescence system. Inflammation.. 1992;16:437-450.[Medline] [Order article via Infotrieve]
41.
McCartney-Francis N, Allen JB, Mizel DE, Albina JE, Xie Q-W, Nathan CF, Wahl SM. Suppression of arthritis by an inhibitor of nitric oxide synthase. J Exp Med.. 1993;178:749-754.
42. Belder AJD, Radomski MW, Why HJF, Richardson PJ, Bucknall CA, Salas E, Martin JF, Moncada S. Nitric oxide synthase activities in human myocardium. Lancet.. 1993;341:84-85.[Medline] [Order article via Infotrieve]
43. Bredt DS, Hwang PS, Snyder SH. Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature.. 1990;347:768-770.[Medline] [Order article via Infotrieve]
44. Schulz R, Nava E, Moncada S. Induction and potential biological relevance of a Ca2+-independent nitric oxide synthase in the myocardium. Br J Pharmacol.. 1992;105:575-580.[Medline] [Order article via Infotrieve]
45.
Roberts AB, Vodouotz Y, Roche NS, Sporn MB, Nathan CF. Role of nitric oxide in antagonistic effects of transforming growth factor- and interleukin-1ß on the beating rate of cultured cardiac myocytes. Mol Endocrinol.. 1992;6:1921-1930.
46. Hiroe M, Ishiyama S, Nishikawa T, Shimojo T, Ito H, Kasajima T, Marumo F. Expression of inducible nitric oxide synthase in the myocardium of acute myocarditis: a serial cardiac biopsy study. J Am Coll Cardiol. 1995;special issue:131A, 726-1.
47.
Drapier J-C, Weizesbin J, Hibbs JB. Interferon- and tumor necrosis factor induce the L-arginine-dependent cytotoxic effector mechanism in murine macrophages. Eur J Immunol.. 1988;18:1587-1592.[Medline]
[Order article via Infotrieve]
48. Dudek RR, Wildhirt S, Conforto A, Pinto V, Suzuki H, Winder S, Bing RJ. Inducible nitric oxide synthase activity in myocardium after myocardial infarction in rabbit. Biochem Biophys Res Commun.. 1994;205:1671-1680.[Medline] [Order article via Infotrieve]
49. Wildhirt SM, Dudek RR, Suzuki H, Pinto V, Narayan KS, Bing RJ. Immunohistochemistry in the identification of nitric oxide synthase isoenzymes in myocardial infarction. Cardiovasc Res.. 1995;29:526-531.[Medline] [Order article via Infotrieve]
50. Steuer DJ, Nathan CF. Nitric oxide: a macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J Exp Med.. 1989;169:1543-1555.
51.
Radi RE, Beckman JS, Bush KM, Freeman BA. Peroxynitrite oxidation of sulfhydryls. J Biol Chem.. 1991;266:4244-4250.
52. Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA. Apparent hydroxyl radical production by perioxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci U S A.. 1990;87:1020-1024.
53. Ishiropoulos H, Zhu L, Chen J, Tsai M, Martin JC, Smith CD, Beckman JS. Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutase. Arch Biochem Biophys.. 1992;298:431-437.[Medline] [Order article via Infotrieve]
54. Huie RE, Padmaja S. The reaction of NO with superoxide. Free Radical Res Commun.. 1992;18:195-199.
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