(Circulation. 2000;101:33.)
© 2000 American Heart Association, Inc.
Clinical Investigation and Reports |
Gene Expression of Antioxidative Enzymes in the Human Heart
Increased Expression of Catalase in the End-Stage Failing Heart
From the Department of Cardiology and Angiology, University Freiburg, Freiburg, Germany (S.D., U.B., K.B.) and the Department of Cardiology and Pneumology, Georg-August-University Goettingen, Germany (G.H., J.P.).
Correspondence to Juergen Prestle, PhD, Klinikum der Georg-August-Universitaet Goettingen, Zentrum Innere Med/Abt Kardiologie & Pneumologie, Robert-Koch-Strasse 40, D-37075 Goettingen, Germany. E-mail prestle{at}med.uni-goettingen.de
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Abstract |
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Background—An increase in oxidative stress is suggested to be intimately involved in the pathogenesis of heart failure. However, gene expression of enzymes that metabolize reactive oxygen metabolites has not been investigated in the human heart.
Methods and Results—Myocardial tissue homogenates of the left ventricular wall from hearts in end-stage failure due to dilated (DCM) or ischemic (ICM) cardiomyopathy (n=12 each), as well as from nonfailing donor hearts (n=12), were analyzed for mRNA levels of manganese superoxide dismutase (MnSOD), copper-zinc superoxide dismutase (CuZnSOD), glutathione peroxidase (GPX), and catalase by Northern blot analyses. Protein levels of MnSOD, CuZnSOD, and catalase were determined by Western blot or ELISA. MnSOD, CuZnSOD, and GPX mRNA levels were similar in all 3 groups. In contrast, catalase mRNA levels were found to be increased by 123±23% in DCM hearts and by 93±10% in ICM hearts (P<0.01 each) compared with control hearts. Likewise, catalase protein levels were found to be increased in failing hearts (DCM by 90±10%, ICM by 90±13%; P<0.05 each) compared with control hearts. In addition, the observed upregulation of catalase mRNA and protein in failing hearts was attended by an increased catalase enzyme activity (DCM by 124±16%, ICM by 117±15%; P<0.01 each), whereas MnSOD, CuZnSOD, and GPX enzyme activity levels were unchanged in failing compared with nonfailing myocardium.
Conclusions—Increased oxidative stress in human end-stage heart failure may result in a specific upregulation of catalase gene expression as a compensatory mechanism, whereas SOD and GPX gene expression remain unaffected.
Key Words: antioxidants • enzymes • free radicals • heart failure • molecular biology
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Introduction |
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Increasing evidence suggests that oxidative stress mediated by the generation of oxygen free radicals plays a critical role in the pathogenesis of heart failure.1 2 3 The myocardium is equipped with a variety of endogenous enzymatic and nonenzymatic antioxidant systems that are sufficient to metabolize oxygen free radicals generated during normal cellular activity. In particular, dismutation of superoxide anion by cytosolic copper/zinc- and mitochondrial manganese-containing superoxide dismutase (CuZnSOD and MnSOD, respectively) and the degradation of H2O2 by glutathione peroxidase (GPX) and catalase limit the cytotoxic effects of reactive oxygen metabolites.4 During pathophysiological conditions, however, the balance between free radicals and antioxidants may shift in favor of a relative increase in free radicals, resulting in increased oxidative stress.
During the last decade, considerable research effort has been directed at the identification of changes in oxidative stress and in antioxidative enzymes as one of the mechanisms underlying the development of heart failure. It has been reported that heart hypertrophy in rats and guinea pigs is associated with a decrease in oxidative stress and an increase in antioxidant reserve,5 6 7 8 whereas heart failure under both acute and chronic conditions is associated with increased oxidative stress and a reduced antioxidant reserve.6 9 10 11 In humans with chronic heart failure, products of free radical reactions, ie, plasma lipid peroxides, are elevated, whereas plasma thiols, as an index of the oxidative status of the extracellular environment, are decreased.2 Expired breath pentane levels, as an index of lipid peroxidation, were found to be elevated in patients with heart failure as well.12 Furthermore, it has been reported that plasma antioxidative enzyme activities were decreased in patients with ischemic heart disease and with heart failure.13 14 However, direct proof for an increased free radical formation in the failing human heart is yet to be furnished.
In addition to the biochemical aspects of oxidative stress, gene expression of antioxidative enzymes in the heart has been investigated during the last several years. In this regard, an increase in myocardial antioxidant enzyme gene expression has been reported after acute oxidative stress induced by endotoxin,15 cytokines,16 17 and ischemia/reperfusion.16 18 However, gene expression of antioxidants in the human heart with end-stage heart failure has not been investigated. In view of increasing evidence for the involvement of oxidative stress in heart failure, it is of considerable interest to examine whether changes in antioxidative enzymes at the transcriptional or translational level may exist under chronic conditions in human heart failure. Therefore, we studied gene expression of CuZnSOD, MnSOD, GPX, and catalase in human end-stage failing hearts from patients with dilated cardiomyopathy (DCM) and ischemic cardiomyopathy (ICM) compared with nonfailing (NF) control hearts.
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Patients
Hearts from patients with end-stage heart failure who were undergoing cardiac transplantation because of either DCM (n=12) or ICM (n=12) were investigated. Furthermore, NF donor hearts (n=12) that were ultimately rejected for transplantation because of technical reasons were also included in this study. Hemodynamic data at the time of cardiac transplantation and age of patients are given in the Table
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The protocol of this study was reviewed and approved by the ethics committee of the University Clinics of Freiburg.
Tissue Sample Preparation
Excised hearts were rinsed immediately in cardioplegic
Krebs-Henseleit solution containing 30 mmol/L 2,3-butanedione
monoxime. Transmural tissue samples derived from the left
ventricular free wall were frozen in liquid nitrogen and
stored at -80°C until use. For Northern blot and Western blot
analyses, as well as for enzyme activity assays, aliquots of
100 mg of tissue were ground in liquid nitrogen and
homogenized with an FP 120 Fast Prep Cell Disrupter (Savant
Instruments) in the respective homogenization
buffer.
Northern Blot Analysis
Myocardial tissue was homogenized in lysis buffer
RTL (Qiagen), and total RNA was extracted by use of an RNeasy
Mini Kit (Qiagen) according to the manufacturer’s instructions. Eight
micrograms of total RNA per lane was size fractionated on a denaturing
1% agarose-formaldehyde gel, transferred to nylon membrane
(Duralon-UV, Stratagene) by overnight capillary blotting, and
immobilized by ultraviolet irradiation. Specific DNA probes
for detection of catalase, MnSOD, CuZnSOD, GPX, and GAPDH gene
transcripts were generated by polymerase chain reaction (PCR) from a
cardiac specific cDNA sample with the following primer pairs: catalase,
forward 5'-TCCGGGATCTTTTTAACGCCATTG-3', reverse
5'-TCGAGCACGGTAGGGACAGTTCAC-3' (nucleotide position 855 to
1216, according to the sequence of the human catalase
gene19 ); MnSOD, forward
5'-CTCCCCGACCTGCCCTACGACTAC-3', reverse
5'-AAACCAAGCCAACCCCAACCTGAG-3' (nucleotide position 176 to
549, according to the sequence of the human SOD2
gene20 ); CuZnSOD, forward
5'-GTGGGGAAGCATTAAAGGACTGAC-3', reverse
5'-CAATTACACCACAAGCCAAACGAC-3' (nucleotide position
160 to 515, according to the sequence of the human SOD1
gene21 ); GPX, forward 5'-GCGGCGGCCCAGTCGGTGTA-3',
reverse 5'-GAGCTTGGGGTCGGTCATAA-3' (nucleotide
position 343 to 759, according to the sequence of the human
GPX1 gene22 ); GAPDH, forward
5'-CCACCCAGAAGACTGTGGAT-3', reverse 5'-GTTGAAGTCAGAGGAGACCACC-3'
(nucleotide position 608 to 921, according to the sequence
of the human GAPDH gene23 ). The identity
of the PCR fragments was verified by sequencing. DNA probes were
labeled with [
32P]dCTP by random priming
(DNA labeling kit, Pharmacia Biotech), and unbound radioactivity was
removed by spin columns. DNA/RNA hybridization was performed with
QuickHyb hybridization solution (Stratagene) for 2 hours at 68°C.
Blots were washed twice in 2xSSC/0.1% SDS for 10 minutes at room
temperature and in 0.1xSSC/0.1% SDS for 30 minutes at 60°C. After
autoradiography, specific signal intensity was
quantified by video-image analysis. GAPDH data were used as an
internal standard to normalize the catalase, MnSOD, CuZnSOD, and GPX
data.
Western Blot Analysis
Protein levels of catalase, MnSOD, and calsequestrin were
examined by Western blot analyses. Tissue samples were
homogenized in ice-cold lysis buffer containing 20
mmol/L Na-HEPES (pH 7.5), 4 mmol/L EGTA, 1% Triton X-100, 2
mmol/L desoxycholate, 1 mmol/L phenylmethylsulfonyl
fluoride, 0.05 mmol/L leupeptin, 1 mmol/L
iodoacetamide, 1 µg/mL aprotinin, and 20 µg/mL trypsin
inhibitor. Crude homogenates were
centrifuged for 5 minutes at 10 000g, and protein
concentration of supernatants was determined by the BCA method (Pierce
Chemical Co). Samples were denatured in electrophoresis buffer
containing 100 mmol/L Tris/Cl (pH 6.8), 8 mmol/L DTT, 2%
SDS, 2% glycerol, and 0.05% bromophenol blue at 95°C and subjected
to SDS-PAGE. Proteins were transferred to nitrocellulose membranes by
electroblotting. Membranes were blocked in 5% nonfat dry milk in
Tris-buffered saline and processed for immunodetection with a rabbit
polyclonal antibody specific for catalase (Paesel 38 Lorei, Hanau,
Germany), a sheep polyclonal antibody specific for MnSOD (Biotrend,
Koeln, Germany), and a rabbit polyclonal antibody specific for
calsequestrin as primary antibodies.24 The secondary
antibodies were peroxidase-conjugated anti-rabbit or anti-sheep
antisera. Visualization of immunoreactive bands was performed with the
enhanced chemoluminescence assay (ECL, Amersham), and signal intensity
was quantified by video-image analysis. The calsequestrin data
were used as an internal standard to normalize the respective catalase
and MnSOD data.
CuZnSOD ELISA
Myocardial protein levels of CuZnSOD were determined with a
commercially available ELISA (CuZnSOD SURALISA, Immundiagnostik GmbH).
Determinations were performed in triplicate according to the
manufacturer’s instructions with the same tissue
homogenates that were used for Western blot
analysis.
Enzyme Activity Assays
Catalase Enzyme Assay
Tissue samples were homogenized in an ice-cold
isotonic 0.01 mol/L sodium phosphate buffer (pH 7.4) and
centrifuged for 5 minutes at 12 000g at 4°C.
Catalase activity was examined in the supernatants by use of a rapid
spectrophotometric method described by Cohen et al.25
Briefly, the catalase-catalyzed decomposition of
H2O2 was determined by
subjecting it to reaction for 3 minutes with a standard excess of
KMnO4 and by subsequent measurement of the
residual KMnO4 at 480 nm. Measurements were
performed in triplicate. Protein concentrations were estimated by the
BCA method. Catalase activity was calculated as units per milligram of
protein.
SOD Enzyme Assay
Tissue samples were homogenized in ice-cold 0.01
mol/L sodium phosphate buffer (pH 7.4) supplemented with 0.03 mol/L KCl
to facilitate the recovery of MnSOD and centrifuged for 5
minutes at 12 000g at 4°C. Total superoxide dismutase
(SOD) activity was examined in the supernatants according to the method
described by Del Maestro and McDonald.26 This assay
is based on the ability of SOD to scavenge superoxide anion radical
(O2-), which decreases the
overall rate of pyrogallol autoxidation. In brief, 1 mL of 0.05 mol/L
Tris-HCL buffer (pH 8.2) containing 1 mmol/L DTPA was added to 40
µL of tissue sample (2 mg of total protein/mL). The reaction was
initiated by the addition of 0.2 mmol/L pyrogallol, and the change
in optical density at 420 nm was recorded for 3 minutes. In a
separate reaction, specific MnSOD activity was measured by the addition
of 1 mmol/L sodium cyanide to the tissue sample to inhibit
CuZnSOD. SOD activity was calculated as units per milligram of protein,
with 1 U of SOD defined as the amount that inhibited the rate of
pyrogallol autoxidation by 50%. We calculated CuZnSOD activity by
subtracting the value using cyanide from the total SOD value.
GPX Enzyme Assay
Tissue homogenates were prepared as described for
the SOD assay. GPX activity was examined in the supernatants according
to the method described by Del Maestro and McDonald.26 The
assay is based on the oxidation of reduced glutathione by GPX coupled
to the disappearance of NADPH by glutathione reductase. In brief, a
40-µL tissue sample (2 mg of protein/mL) was added to 0.9 mL of 0.05
mol/L potassium phosphate buffer (pH 7.0) with 0.5 mmol/L DTPA
containing 2 mmol/L glutathione (Sigma), 1 U of glutathione
reductase (Sigma), and 0.16 mmol/L NADPH (Sigma). After addition
of 0.6 mmol/L t-butyl hydroperoxide, the change in
optical density at 340 nm was recorded for 3 minutes. GPX activity
was calculated as units per gram of protein, with 1 U of GPX causing
the oxidation of 1 µmol/L glutathione per minute in the system
outlined.
Statistical Analysis
All experimental values represent the mean of at least 2
independent determinations. Data are expressed as mean±SEM. For
statistical analyses, median values of the 3 study groups (NF,
DCM, and ICM) were compared by Kruskal-Wallis 1-way ANOVA on ranks,
followed by a multiple comparison procedure (Dunn’s method) to isolate
study groups that differed from the others. A value of
P<0.05 was considered statistically significant. For
graphical reasons, NF data from Northern and Western blots were set to
100%. DCM and ICM data are expressed in percent of NF data.
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Results |
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Relative mRNA Abundance of Antioxidative Enzymes
Autoradiograms shown in Figure 1A
indicate that specific cDNA probes for
human CuZnSOD recognized a major 0.7-kb and a minor 0.9-kb transcript,
a single 4-kb transcript for human MnSOD, and a single 1-kb transcript
for human GPX, respectively, in all hearts examined. The relative
mRNA levels, measured as the ratios of the intensities of mRNA
for antioxidative enzymes to those of GAPDH mRNA, are shown in Figure 1B. For CuZnSOD, MnSOD, and GPX mRNA contents, no
significant differences could be detected between the 3 study
groups.
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The autoradiogram in Figure 2A indicates that the cDNA probe for
human catalase recognized a single 2.5-kb transcript. In contrast to
the other antioxidative enzymes examined, relative catalase mRNA
contents were significantly elevated in DCM hearts (Figure 2B;
123±23% increase compared with NF; P<0.01) and in ICM
hearts (93±10% increase compared with NF; P<0.01). No
statistically significant difference was found between the DCM and ICM
groups.
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Protein Contents of Antioxidative Enzymes
Because mRNA levels may not necessarily reflect the corresponding
protein levels, we also investigated protein contents of catalase and
MnSOD by Western blot analyses. The antibodies for catalase and
MnSOD specifically detected proteins with molecular sizes of 58 kDa for
catalase and 21 kDa for MnSOD, respectively. To account for potential
differences in extracellular matrix content in tissue
homogenates from failing and nonfailing
myocardium, protein data represent relative values
normalized to the myocyte-specific protein calsequestrin, for which
equal distribution in NF, DCM, and ICM hearts was reported in former
studies24 and which could also be confirmed in the
present study (data not shown). CuZnSOD protein contents determined
by ELISA represent absolute values and are given in nanograms
per milligram of total protein. Determination of human GPX protein
level was not done in the present study, because the tested
commercially available antibodies against GPX detected proteins of
different molecular weights than the reported size for the human GPX
protein.27
Analyses of antioxidative enzyme protein levels revealed
similar results as for the respective mRNA levels. As shown in Figure 3, catalase protein contents normalized
to calsequestrin were significantly elevated in DCM and ICM hearts
compared with NF controls (DCM 90±10% increase and ICM 90±13%
increase, P<0.05 each), whereas MnSOD (Figure 4A) and CuZnSOD (Figure 4B)
protein levels showed no significant differences between failing and
nonfailing myocardium.
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We normalized protein levels of catalase and MnSOD to total protein content in crude homogenates in addition to normalizing them to calsequestrin. Expressed in relative arbitrary units, protein levels of MnSOD were 1.03±0.03 in NF myocardium, 1.05±0.05 in DCM hearts, and 0.96±0.09 in ICM hearts, with no statistically significant difference between groups. Catalase protein levels were increased in DCM and ICM hearts by 72.9±7.0% and 70.4±7.1% compared with NF hearts, respectively (P<0.05 each; 0.58±0.12 versus 1.01±0.09 and 0.83±0.08 relative arbitrary units in NF, DCM, and ICM hearts, respectively).
Enzyme Activity Levels
In accordance with mRNA and protein data, catalase activity was
found to be increased in DCM and ICM hearts by 124±16% and 117±15%,
respectively (P<0.01 each), compared with NF controls
(Figure 5A). Enzyme activity levels of
MnSOD, CuZnSOD, and GPX were similar in all 3 groups (Figure 5, B through D).
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Discussion |
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There is ample support for the hypothesis that an increased generation of oxygen free radicals is involved in the pathogenesis as well as the progression of heart failure. It has been reported that in various animal models of heart failure, an increase in oxidative stress may be combined with a decrease in scavenging enzyme activity, either as a cause or as a result.5 6 7 8 However, it remains unclear whether human end-stage heart failure is also associated with an altered myocardial antioxidant reserve.
We studied gene expression and enzyme activity levels of myocardial
scavenger enzymes in human hearts with end-stage heart failure of
different origins (DCM and ICM) compared with NF hearts. The results of
our study clearly demonstrate that no differences in gene expression of
MnSOD, CuZnSOD, and GPX exist between failing and NF human hearts.
On the contrary, catalase gene expression was 2-fold higher in the
failing heart than in the NF heart. We found catalase expression to be
increased on both the mRNA and protein level, with a conformable
increase in catalase enzyme activity as well, whereas enzyme activities
of MnSOD, CuZnSOD, and GPX were similar between NF and end-stage
failing myocardium.
These findings are interesting under various aspects. It has been suggested that SOD and GPX play the major role in detoxification of reactive oxygen metabolites in the heart.28 The unchanged gene expression of MnSOD, CuZnSOD, and GPX in human end-stage failing myocardium demonstrated in the present study is in contrast with the findings of several animal studies and the report about a significant reduction in plasma scavenging enzyme activities in patients with heart failure,14 although plasma and tissue levels of antioxidative enzymes cannot necessarily be compared. Our data indicate that the myocardial antioxidant reserve is not diminished during human end-stage heart failure, and a deficit in antioxidative capacity may therefore not play a significant pathophysiological role in human heart failure.
Interestingly, expression and activity of catalase were found to be considerably increased in the failing myocardium. Although SOD is the first line of defense against oxygen free radical–mediated damage, it acts to increase the levels of H2O2 by virtue of catalyzing the dismutation of superoxide anion to H2O2. The major danger of H2O2 accumulation is the production of highly reactive hydroxyl radical, for which no physiological defense system exists.4 As a result, catalase and GPX become the most crucial antioxidative enzymes, because they act to detoxify H2O2. It was shown that the heart contains <2% of the catalase found in liver, yet it produces a greater amount of hydrogen peroxide per gram of tissue than any other organ.29 30 31 Together with the fact that catalase has a lower affinity for H2O2, it is therefore believed that the glutathione redox system acts as the major route for the metabolism of H2O2 in the heart and that catalase is of little importance.32 However, catalase enables the cell to decompose H2O2 regardless of the cellular concentration of glutathione, which may be important in light of evidence that high plasma levels of tumor necrosis factor, as found in patients with heart failure, reduce tissue glutathione levels.33 34 Thus, catalase may be of particular relevance in the failing myocardium. Furthermore, a number of studies do suggest a significant role for endogenous catalase, and catalase mRNA is reported to be upregulated in the mammalian heart after subjection to a stress insult.17 35 For example, it is well known that reperfusion after ischemia causes generation of oxygen free radicals and induces oxidative stress. In this regard, it has been shown that repeated ischemia and reperfusion enhanced catalase expression,17 and there is evidence that in addition to heat shock proteins, catalase is implicated in the cardioprotective effect of heat stress against reperfusion arrhythmia.36 Furthermore, pretreatment with catalase but not SOD prevented abnormalities in contraction-relaxation processes and offered essentially complete functional protection against oxygen-derived free radicals in rat papillary muscle preparations.37 Protection against a lethal oxidant injury of H2O2 has also been demonstrated to be conferred by adenovirus-mediated gene transfer of human catalase into human umbilical vein endothelial cells.38 Finally, it has been found that under chronic oxidative stress by direct challenge of myocytes with H2O2, catalase but not GPX was selectively induced by transcriptional activation.39
In this context, it should be mentioned that we only analyzed gene expression and enzyme activity levels of cellular antioxidative enzymes. Expression and enzyme activity levels of extracellular SOD (EC-SOD), the third SOD isozyme, were not determined in the present study. Transgene overexpression of EC-SOD was shown to provide rabbit hearts with substantial protection against myocardial stunning without concomitant administration of catalase and to preserve myocardial function after ischemia-reperfusion injury in isolated murine hearts.40 41 We cannot exclude that plasma levels or tissue vascular levels of EC-SOD are altered during end-stage heart failure in response to an increase in oxidative stress. For example, EC-SOD expression and activity were shown to be reduced in advanced human atherosclerotic lesions.42 However, Adachi et al43 reported similar plasma concentrations of EC-SOD in healthy individuals compared with patients with heart diseases, although only a limited number of patients with unspecified heart disease were included in that study. Furthermore, because the present study was performed in myocardial tissue samples, we cannot differentiate whether catalase was specifically increased in the myocardium, in the microvascular system, or in both. Likewise, an increased expression of SOD and GPX in cardiomyocytes could be offset by a reduced expression within the vascular system.
In summary, it appears that an increase in endogenous catalase can be regarded as an effort made by the heart to protect itself from an oxidative assault. Under physiological conditions, catalase expression seems to be much lower than with other antioxidative enzymes. However, the results of the present study underscore the importance of catalase induction at the transcriptional level as an adaptive cardioprotective response under chronic pathophysiological conditions.
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Acknowledgments |
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This work was supported by the Deutsche Forschungsgemeinschaft, grant HA 1233/3-3.
Received May 12, 1999; revision received August 4, 1999; accepted August 5, 1999.
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References |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Singh N, Dhalla AK, Seneviratne C, Singal PK. Oxidative stress and heart failure. Mol Cell Biochem. 1995;147:77–81.[Medline] [Order article via Infotrieve]
2.
Belch JJF, Bridges AB, Scott N, Chopra M. Oxygen free
radicals and congestive heart failure. Br Heart J. 1991;65:245–248.
3. Hoeschen RJ. Oxidative stress and cardiovascular disease. Can J Cardiol. 1997;13:1021–1025.[Medline] [Order article via Infotrieve]
4. Hess ML, Manson NH. The oxygen free radical system and myocardial dysfunction. Adv Myocardiol.. 1985;5:177–181.[Medline] [Order article via Infotrieve]
5.
Dhalla AK, Singal PK. Antioxidant changes in
hypertrophied and failing guinea pig hearts. Am J
Physiol. 1994;266:H1280–H1285.
6. Dhalla AK, Hill MF, Singal PK. Role of oxidative stress in transition of hypertrophy to heart failure. J Am Coll Cardiol. 1996;28:506–514.[Abstract]
7.
Gupta M, Singal PK. Higher antioxidative capacity
during a chronic stable heart hypertrophy. Circ
Res. 1989;64:398–406.
8.
Kirshenbaum LA, Singal PK. Increase in
endogenous antioxidant enzymes protects hearts against
reperfusion injury. Am J Physiol. 1993;265:H484–H493.
9.
Hill MF, Singal PK. Right and left myocardial
antioxidant responses during heart failure subsequent to myocardial
infarction. Circulation. 1997;96:2414–2420.
10. Khaper N, Singal PK. Effects of afterload-reducing drugs on pathogenesis of antioxidant changes and congestive heart failure in rats. J Am Coll Cardiol. 1997;29:856–861.[Abstract]
11. Hill MF, Singal PK. Antioxidant and oxidative stress changes during heart failure subsequent to myocardial infarction in rats. Am J Pathol. 1996;148:291–300.[Abstract]
12. Sobotka PA, Brottman MD, Weitz Z, Birnbaum AJ, Skosey JL, Zarling EJ. Elevated breath pentane in heart failure reduced by free radical scavenger. Free Radic Biol Med. 1993;14:643–647.[Medline] [Order article via Infotrieve]
13. Chandra M, Chandra N, Agrawal R, Kumar A, Ghatak A, Pandey VC. The free radical system in ischemic heart disease. Int J Cardiol. 1994;43:121–125.[Medline] [Order article via Infotrieve]
14. Ghatak A, Brar MJS, Agarwal A, Goel N, Rastogi AK, Vaish AK, Sircar AR, Chandra M. Oxy free radical system in heart failure and therapeutic role of oral vitamin E. Int J Cardiol. 1996;57:119–127.[Medline] [Order article via Infotrieve]
15. Ghosh B, Hanevold CD, Dobashi K, Orak JK, Singh I. Tissue differences in antioxidant enzyme gene expression in response to endotoxin. Free Radic Biol Med. 1996;21:533–540.[Medline] [Order article via Infotrieve]
16. Chandrasekar B, Colston JT, Freeman GL. Induction of proinflammatory cytokine and antioxidant enzyme gene expression following brief myocardial ischemia. Clin Exp Immunol.. 1997;108:346–351.[Medline] [Order article via Infotrieve]
17. Das DK, Maulik N, Moraru II. Gene expression in acute myocardial stress: induction by hypoxia, ischemia, reperfusion, hyperthermia and oxidative stress. J Mol Cell Cardiol. 1995;27:181–193.[Medline] [Order article via Infotrieve]
18.
Das DK, Engelman RM, Kimura Y. Molecular adaptation of
cellular defences following preconditioning of the heart by repeated
ischemia. Cardiovasc Res. 1993;27:578–584.
19. Bell GI, Najarian RC, Mullenbach GT, Hallewell RA. cDNA sequence coding for human kidney catalase. Nucleic Acids Res. 1986;14:5561–5562.
20. Ho YS, Crapo JD. Isolation and characterization of complementary DNAs encoding human manganese-containing superoxide dismutase. FEBS Lett. 1988;229:256–260.[Medline] [Order article via Infotrieve]
21.
Hallewell RA, Masiarz FR, Najarian RC, Puma JP, Quiroga
MR, Randolph A, Sanchez-Pescador R, Scandella CJ, Smith B, Steimer KS,
Mullenbach GT. Human Cu/Zn superoxide dismutase cDNA: isolation of
clones synthesizing high levels of active or inactive enzyme from an
expression library. Nucleic Acids Res. 1985;13:2017–2034.
22.
Sukenaga Y, Ishida K, Takeda T, Takagi K. cDNA sequence
coding for human glutathione peroxidase. Nucleic Acids Res. 1987;15:7178.
23.
Tokunaga K, Nakamura Y, Sakata K, Fujimori K, Ohkubo M,
Sawada K, Sakiyama S. Enhanced expression of a
glyceraldehyde-3-phosphate dehydrogenase gene in human
lung cancers. Cancer Res. 1987;47:5616–5619.
24.
Meyer M, Schillinger W, Pieske B, Holubarsch C,
Heilmann C, Posival H, Kuwajima G, Mikoshiba K, Just H, Hasenfuss G.
Alterations of sarcoplasmic reticulum proteins in failing human dilated
cardiomyopathy. Circulation. 1995;92:778–784.
25. Cohen G, Dembiec D, Marcus J. Measurement of catalase activity in tissue extracts. Anal Biochem. 1970;34:30–38.[Medline] [Order article via Infotrieve]
26. Del Maestro RF, McDonald W. Oxidative enzymes in tissue homogenates. In: Greenwald RA, ed. Handbook of Methods for Oxygen Radical Research. Boca Raton, Fla: CRC Press; 1985:291–296.
27. Rey C, Vericel E, Nemoz G, Chen W, Chapuy P, Lagarde M. Purification and characterization of glutathione peroxidase from human blood platelets: age-related changes in the enzyme. Biochim Biophys Acta. 1994;1226:219–224.[Medline] [Order article via Infotrieve]
28. Doroshow JH, Locker GY, Myers CE. Enzymatic defenses of the mouse heart against reactive oxygen metabolites. J Clin Invest. 1980;65:128–135.
29.
Marklund L, Westman G, Lundgren E, Roos G. CuZn
superoxide dismutase, Mn superoxide dismutase, catalase and glutathione
peroxidase in normal and neoplastic cell lines and normal human tissue.
Cancer Res. 1982;42:1955–1961.
30.
Chance B, Sies H, Boveris A. Hydroperoxide
metabolism in mammalian organs. Physiol Rev. 1979;59:527–605.
31. Lai C-C, Huang W-H, Askari A, Klevay LM, Chiu TH. Expression of glutathione peroxidase and catalase in copper-deficient rat liver and heart. Nutr Biochem. 1995;6:256–262.
32.
Janssen M, Van DeMeer P, DeJong JW. Antioxidant
defences in rat, pig, guinea pig, and human hearts: comparison with
xanthine oxidoreductase activity. Cardiovasc Res.. 1993;27:2052–2057.
33.
Ishii Y, Partridge CA, Del Vecchio PJ, Malik AB. Tumor
necrosis factor--mediated decrease in glutathione increases the
sensitivity of pulmonary vascular endothelial
cells to H2O2. J
Clin Invest. 1992;89:794–802.
34.
Phelps DT, Ferro TJ, Higgins PJ, Shankar R, Parker DM,
Johnson A. TNF- induces peroxynitrite-mediated depletion of
lung endothelial glutathione via protein kinase
C. Am J Physiol.. 1995;269:L551–L559.
35. Steare SE, Yellon DM. The potential for endogenous myocardial antioxidants to protect the myocardium against ischemia-reperfusion injury: refreshing the parts exogenous antioxidants cannot reach? J Mol Cell Cardiol. 1995;27:65–74.[Medline] [Order article via Infotrieve]
36. Joyeux M, Ribuot C, Bourlier V, Verdetti J, Durand A, Richard M-J, Godin-Ribout D, Demenge P. In vitro antiarrhythmic effect of prior whole body hyperthermia: implication of catalase. J Mol Cell Cardiol. 1997;29:3285–3292.[Medline] [Order article via Infotrieve]
37. Blaustein AS, Schine L, Brooks WW, Fanburg BL, Bing OHL. Influence of exogenously generated oxidant species on myocardial function. Am J Physiol. 1986;250:H595–H599.
38.
Erzurum SC, Lemarchand P, Rosenfeld MA, Yoo J-H,
Crystal RG. Protection of human endothelial cells from
oxidant injury by adenovirus-mediated transfer of human catalase cDNA.
Nucleic Acids Res. 1993;21:1607–1612.
39. Lai C-C, Peng M, Huang L, Huang W-H, Chiu TH. Chronic exposure of neonatal cardiac myocytes to hydrogen peroxide enhances the expression of catalase. J Mol Cell Cardiol.. 1996;28:1157–1163.[Medline] [Order article via Infotrieve]
40.
Li Q, Bolli R, Qiu Y, Tang XL, Murphree SS, French BA.
Gene therapy with extracellular superoxide dismutase attenuates
myocardial stunning in conscious rabbits. Circulation. 1998;98:1438–1448.
41. Chen EP, Bittner HB, Davis RD, Folz RJ, Van Trigt P. Extracellular superoxide dismutase transgene overexpression preserves postischemic myocardial function in isolated murine hearts. Circulation. 1996;94(suppl II):II-412–II-417.
42.
Luoma JS, Strålin P, Marklund SL, Hiltunen TP,
Särkioja T, Ylä-Herttuala S. Expression of extracellular
SOD and iNOS in macrophages and smooth muscle cells in human
and rabbit atherosclerotic lesions. Arterioscler Thromb Vasc
Biol. 1998;18:157–167.
43. Adachi T, Nakamura M, Yamada H, Futenma A, Kato K, Hirano K. Quantitative and qualitative changes of extracellular-superoxide dismutase in patients with various diseases. Clin Chim Acta. 1994;229:123–131.[Medline] [Order article via Infotrieve]
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