(Circulation. 2001;103:1695.)
© 2001 American Heart Association, Inc.
Basic Science Reports |
Hmox-1 Constitutes an Adaptive Response to Effect Antioxidant Cardioprotection
A Study With Transgenic Mice Heterozygous for Targeted Disruption of the Heme Oxygenase-1 Gene
From the Cardiovascular Research Center, University of Connecticut School of Medicine, Farmington (T.Y., N.M., D.K.D.); the Institute of Chemical Toxicology, Wayne State University, Detroit, Mich (Y.-S.H.); and the Department of Molecular Genetics, Louisiana University Medical Center, New Orleans (J.A.).
Correspondence to Dipak K. Das, PhD, University of Connecticut, School of Medicine, Farmington, CT 06030-1110. E-mail ddas{at}neuron.uchc.edu
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Abstract |
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Background—Heme oxygenase-1 (Hmox-1) has been implicated in protection of cells against ischemia/reperfusion injury.
Methods and Results—To examine the physiological role of Hmox-1, a line of heterozygous Hmox-1-knockout mice was developed by targeted disruption of the mouse Hmox-1 gene. Transgene integration was confirmed and characterized at the protein level. A 40% reduction of Hmox-1 protein occurred in the hearts of Hmox-1+/- mice compared with those of wild-type mice. Isolated mouse hearts from Hmox-1+/- mice and wild-type controls perfused via the Langendorff mode were subjected to 30 minutes of ischemia followed by 120 minutes of reperfusion. The Hmox-1+/- hearts displayed reduced ventricular recovery, increased creatine kinase release, and increased infarct size compared with those of wild-type controls, indicating that these Hmox-1+/- hearts were more susceptible to ischemia/reperfusion injury than wild-type controls. These results also suggest that Hmox-1+/- hearts are subjected to increased amounts of oxidative stress. Treatment with 2 different antioxidants, Trolox or N-acetylcysteine, only partially rescued the Hmox-1+/- hearts from ischemia/reperfusion injury. Preconditioning, which renders the heart tolerant to subsequent lethal ischemia/reperfusion, failed to adapt the hearts of the Hmox-1+/- mice compared with wild-type hearts.
Conclusions—These results demonstrate that Hmox-1 plays a crucial role in ischemia/reperfusion injury not only by functioning as an intracellular antioxidant but also by inducing its own expression under stressful conditions such as preconditioning.
Key Words: heme oxygenase • genes • oxygen • stress • ischemia
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Introduction |
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Heme oxygenase (HO) catalyzes the reaction for heme metabolism, yielding equimolar quantities of CO, iron, and biliverdin.1 The HO system consists of 2 isoforms: oxidative stress–inducible HO-1 (Hmox-1), also known as heat-shock protein (HSP) 32, and the constitutive isozyme HO-2.2 In addition to oxidants, a variety of other environmental stresses, including heat stress, hypoxia, metals, endotoxin, and certain hormones, can induce Hmox-1.3 4
Recent evidence suggests that Hmox-1 induction plays a role in cellular protection against injury caused by the reactive oxygen species (ROS).5 Enhancement of HO activity by endotoxin attenuated renal failure, and this protection was lost when HO was inhibited with protoporphyrin.6 In another study, Hmox-1 mediated an adaptive response to oxidative stress in skin fibroblasts.7 ROS produced in the ischemic/reperfused myocardium has been shown to induce Hmox-1 gene expression.8
The physiological significance of Hmox-1 induction during myocardial ischemia remains unknown. It seems reasonable to speculate that Hmox-1 induction during ischemia/reperfusion is the heart’s own stress signal for survival against oxidative stress. Although a role of Hmox-1 in myocardial protection has been speculated upon, a definitive cardioprotective role of Hmox-1 is lacking. To fill this gap, we developed an Hmox-1+/- mouse by targeted disruption of the mouse Hmox-1 gene. The isolated hearts with 1 functional copy of the Hmox-1 gene and those of matched wild-type mice were subjected to ischemia/reperfusion. To examine whether Hmox-1 plays a role in stress adaptation, the hearts were subjected to preconditioning by cyclic episodes of ischemia/reperfusion. The results demonstrated impaired ventricular recovery and increased infarct size for the Hmox-1+/- hearts compared with those from wild-type controls, suggesting that Hmox-1+/- hearts are vulnerable to ischemia/reperfusion injury. Improvement of cardiac function was less apparent for the Hmox-1+/- mice after preconditioning.
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Methods |
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Targeted Disruption of the Mouse Hmox-1 Gene
As shown in Figure 1
, the XhoI
fragment of the mouse Hmox-1
gene9 spanning from part of
exon 1 to intron 2 was cloned into the
XhoI site of the targeting
vector pPNT.10 The 3'
XbaI-EcoRI
fragment of the gene-containing sequence of intron 3 to 3' flanking
region of the gene was subcloned into the corresponding
XbaI and
EcoRI sites in pPNT. The
targeting vector was linearized with
NotI digestion and then
transfected into R1 embryonic stem (ES)
cells11 by electroporation.
Five homologous recombinant ES clones were identified from 224 clones
by DNA blot analysis using the 5' external probe encompassing
sequences from nucleotide -133 to +73. Male chimeric
mice, generated by microinjecting C57BL/6 blastocytes with 3
independent clones, were used in breeding with C57BL/6 female mice.
Germ-line transmission of 129/Sv chromosomes was achieved by chimeric
mice derived from each of the homologous recombinant ES clones. Line
183
Hmox-1+/-
mice were used in this study.
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Protein Blot Analysis of
Hmox-1 in Hearts
Hearts were homogenized in 50 mmol/L
phosphate buffer, pH 7.8, containing 0.1% Triton X-100 with a Polytron
homogenizer. The homogenates were
centrifuged at 20 000g
and stored at -70°C. Protein contents were determined with an assay
kit (Pierce).
For protein blot analysis, total cellular protein was separated on a 12% SDS-polyacrylamide gel and electrophoretically transferred onto a nitrocellulose paper. The protein blot filter paper was incubated in 1x HBSS plus a 1:2000 dilution of a monoclonal anti-human Hmox-1 antibody (StressGen) for 2 hours. The filter paper was washed with HBSS containing 1% nonfat milk, incubated with a 1:3000 dilution of horseradish peroxidase–conjugated goat anti-mouse IgG (Bio-Rad, Hercules) in HBSS plus 5% nonfat milk for 1 hour, washed again with HBSS, and then subjected to autoradiography in the presence of an enhanced chemiluminescent substrate (Pierce). Western blot data were quantified by densitometric scanning.
Measurement of Contractile Function
Twenty-four mice (10 to 12 weeks old) were divided
into 2 groups:
Hmox-1+/-
(n=12) and wild-type (n=12). They were anesthetized with
pentobarbital (200 mg/kg). The heart was excised, the aorta cannulated,
and the heart perfused with Krebs-Henseleit buffer (KHB) by the
Langendorff method.12 The
effluent was collected before ischemia and during reperfusion
and stored at -20°C for subsequent measurement of creatine kinase
(CK) and malondialdehyde (MDA). A 4-0 silk suture on a round-bodied
needle was passed through the apex of the heart and attached to the
apex, which in turn was attached to a force transducer. The heart rate,
force developed by the heart (DF), and first derivative of developed
force (dF/dt) were
recorded.12 The data on
myocardial contractile function were recorded and analyzed
in real time with the Cordat II data acquisition and
presentation system (Triton Technologies). The hearts were
subjected to 30 minutes of ischemia by clamping of the aortic
cannula, followed by 2 hours of reperfusion.
Antioxidant Therapy
To examine whether antioxidants could protect the
Hmox-1+/-
hearts from ischemia/reperfusion injury, 2 different protocols
were used. (1) For in vivo studies, both wild-type and
Hmox-1+/-
mice were injected once every 24 hours with either Trolox (10 mg/kg
IV), an analogue of vitamin E, or
N-acetylcysteine (NAC, 1.5
mg/kg IV), a precursor of glutathione synthesis and an antioxidant.
Control hearts were injected with saline only. (2) In vitro experiments
were performed by preperfusion of the isolated wild-type and
Hmox-1+/-
hearts for 15 minutes with KHB buffer in the presence or absence of
Trolox (10 mmol/L) or NAC (10 mmol/L). All hearts were
subjected to 30 minutes of ischemia followed by 2 hours of
reperfusion. The cardioprotective abilities of the antioxidants were
examined by study of ventricular function, infarct size,
and CK release.
Preconditioning Protocol
Another group of hearts from
Hmox-1+/-
and wild-type mice were preconditioned by subjection to 3 cycles of 5
minutes of ischemia each, followed by another period of 5
minutes of
reperfusion.13 14
This preconditioning protocol has been found to render the heart
tolerant to the subsequent ischemic stress. All hearts were
subjected to 30 minutes of ischemia followed by 2 hours of
reperfusion.
Estimation of CK Release
CK release from the heart was estimated in the
perfusate collected from the heart by use of a CK assay kit
(Sigma). The results are expressed as IU/L of total CK release from the
heart.
Estimation of MDA
MDA was assayed in the coronary
perfusate as described
previously15 to monitor the
development of oxidative stress. The MDA was derivatized with
DNPH.15 Aliquots of 25 µL
of derivatized MDA in acetonitrile were injected onto a Beckman
Ultrasphere C18 (3-mm) column in a Waters
high-performance liquid chromatograph (Waters Corp).
The products were eluted isocratically and detected at 307, 325,
and 356 nm. The amount of MDA was quantified by use of a Maxima
software program (Waters).
Measurement of Infarct Size
At the end of reperfusion, a 10% (wt/vol) solution
of triphenyl tetrazolium in phosphate buffer was infused into the
aortic cannula.16 The hearts
were excised and stored at -70°C. Sections (0.8 mm) of frozen
heart were fixed in 2% paraformaldehyde, placed
between 2 coverslips, and digitally imaged with a Microtek ScanMaker
600z. To quantify the areas of interest in pixels, an NIH Image 5.1 (a
public-domain software package) was used. The infarct size (transmural)
was quantified in pixels.
Statistical Analysis
For statistical analysis, a 2-way ANOVA
followed by Scheffé’s test was first carried out to test for any
differences between groups. If differences were established, the values
were compared by Student’s t
test for paired data. The values were expressed as mean±SEM. The
results were considered significant at a level of
P<0.05.
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Results |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Generation and Characterization of Hmox-1+/- Mice
As shown in Figure 1a
, the mouse Hmox-1 gene was
inactivated by replacement of exon 2 with a neomycin
resistance (neo) cassette. Insertion of the neo gene creates a new
PstI restriction site in the
Hmox-1 gene, resulting in a shorter
PstI genomic fragment from the
targeted allele than that from the wild-type allele
(Figure 1b). Heterozygous
Hmox-1+/-
mice were apparently healthy and showed no pathological or phenotypic
differences from the wild-type littermates. No homozygous knockout mice
could be detected in the progeny from heterozygote intercross, however,
indicating that embryonic lethality occurs in the homozygous mice. A
40% reduction of Hmox-1 was evident in the
heart of the heterozygous mouse compared with that of the wild-type
mouse
(Figure 1c). We reasoned that a decrease in expression of
Hmox-1 protein due to the targeted mutation in
Hmox-1+/-
mice might render the mice more susceptible to an increased oxidative
stress than controls.
Myocardial Performance
All hearts recovered their beats spontaneously after 30
minutes of ischemia. The heart rates remained slightly but not
significantly lowered, however, compared with baseline (results not
shown). No significant difference was found in heart rate between
groups throughout the experiment. Coronary flow progressively
lowered during the reperfusion, but differences were not significant
(Figure 2). The DF during reperfusion was lower for the
Hmox-1+/-
mouse hearts than wild-type hearts
(Figure 3, top). In control hearts, DF recovered to near
baseline levels after 90 minutes of reperfusion. Conversely, DF did not
recover beyond 87% of baseline for the
Hmox-1+/-
hearts. At all points, DF showed significantly lower recovery for
Hmox-1+/-
hearts than wild-type hearts after 30 minutes of reperfusion
(P<0.05). A similar trend was
seen in dF/dtmax
(Figure 3, bottom). A significantly lower recovery of
dF/dtmax occurred in
Hmox-1+/-
hearts after 30 minutes of reperfusion
(P<0.05). These results
demonstrate that
Hmox-1+/-
mouse hearts showed significantly lower contractile recovery after 30
minutes of ischemia.
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), wild-type preconditioned (•), Hmox-1+/- (
), and Hmox-1+/- preconditioned (
) hearts. Results are expressed as mean±SEM for n=12 per group.
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Preconditioning rendered the hearts of the wild-type mice tolerant to ischemia/reperfusion injury, as evidenced by improved DF and dF/dt compared with wild-type hearts not subjected to preconditioning. In contrast, the preconditioned hearts of the Hmox-1 mice did not exhibit similar improvement in functional recovery, and there were no significant differences of DF and dF/dt between preconditioned and nonpreconditioned hearts.
Both in vivo and in vitro, Trolox and NAC treatments
partially improved the contractile function. For example,
postischemic DF and dF/dt were improved significantly but
only partially compared with those of
Hmox-1+/-
hearts
(Table).
Interestingly, both antioxidants (in vivo or in vitro) improved DF and
dF/dt to the same extent. Baseline values of the antioxidant-treated
(in vivo or in vitro) and untreated groups were
identical.
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CK Release From Heart
Total CK release from the heart
(Figure 4) was negligible for all the groups before
ischemia. After ischemia, CK release increased in both
groups, but the amount of release was much higher for
Hmox-1+/-
hearts. At 30 minutes of reperfusion, CK release was 70±9.7 IU/mL for
Hmox-1+/-
mice versus 50±11.5 IU/mL
(P<0.05) for nontransgenic
controls. At 60 minutes of reperfusion, CK release was 120±8.7 IU/mL
for
Hmox-1+/-
hearts versus 82±9.4 IU/mL
(P<0.05) for nontransgenic
controls. Similarly, after 120 minutes of reperfusion, CK release from
the knockout mouse hearts amounted to 239±8.1 IU/mL compared with
135±7.4 IU/mL for controls.
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P<0.05 vs Hmox-1+/-.Preconditioning significantly reduced the amount of CK in wild-type hearts compared with the nonpreconditioned group. The amount of CK release was also less for the preconditioned Hmox-1+/- hearts than for nonpreconditioned hearts, but the differences did not reach significance except at 90 and 120 minutes of reperfusion.
In vivo and in vitro, Trolox and NAC treatments partially
but significantly reduced the CK release compared with that from the
Hmox-1+/-
hearts
(Table).
Again, both antioxidants reduced the CK release to the same extent.
Baseline values of the antioxidant-treated and untreated groups were
identical.
Myocardial Infarction
The portion of the heart not stained by tetrazolium was
measured and calculated as the infarct area. Mean value of infarct size
in the knockout group was significantly higher than that in the
wild-type group (48.5±3.2% versus 28.5±2.4% for wild-type,
P<0.05)
(Figure 5). Our results indicated that
Hmox-1+/-
hearts had significantly higher myocardial necrosis.
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A reduced amount of infarct size was apparent in the hearts subjected to preconditioning. Significantly less infarct was noticed, however, for the preconditioned wild-type hearts than nonpreconditioned hearts.
Both in vivo and in vitro, Trolox and NAC treatments
partially but significantly reduced the infarct size compared with the
Hmox-1+/-
hearts
(Table).
Similar to the previous results, both antioxidants reduced the infarct
size to the same extent.
MDA Formation
MDA, measured as MDA-DNPH, increased
progressively during reperfusion for the
nonpreconditioned hearts
(Figure 6). The amount of MDA production, however,
was significantly higher at all points in the
Hmox-1+/-
hearts than in control hearts, demonstrating that
Hmox-1+/-
hearts were subjected to increased amounts of oxidative stress.
Preconditioning reduced the amount of MDA formation in both groups of
hearts. A significantly higher amount of MDA was found in the
Hmox-1 hearts than in the wild-type control
hearts.
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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As mentioned earlier, Hmox-1+/- mouse hearts showed
40%
reduction of Hmox-1 protein compared with
Hmox-1+/+ hearts. The
activities of other key antioxidant enzymes, including manganese
superoxide dismutase (Mn-SOD), catalase, glutathione peroxidase,
glutathione reductase, and glucose-6-phosphate-dehydrogenase, remained
unaltered in the
Hmox-1+/-
hearts compared with wild-type controls (data not shown). The results
thus truly reflect the effects of
Hmox-1+/-
in myocardial ischemia/reperfusion injury. Hmox-1+/- hearts were more susceptible to ischemia/reperfusion injury, as evidenced by greater infarct size, increased CK release, and reduced ventricular recovery compared with wild-type controls. In addition, these Hmox-1+/- hearts could not be preconditioned like the hearts of the wild-type controls, suggesting that Hmox-1 plays a role in the adaptation to stress. This is further supported by the results with MDA, because MDA formation, a presumptive marker for oxidative stress, was higher for the preconditioned Hmox-1+/- mouse hearts than the wild-type hearts.
During the course of our studies, Poss and Tonegawa17 18 have reported generation of Hmox-1+/- mice in which exons 3, 4, and part of 5 were replaced by the neo cassette. Whereas they were able to obtain homozygous knockout mice from heterozygous breeding at a lower-than-expected frequency, an indication of embryonic lethality due to Hmox-1 deficiency, we could not detect any homozygous knockout mice in our breeding colony. The discrepancy in our breeding results is not understood. It may result from the fact that different ES cells were being used in each laboratory. The studies by Poss and Tonegawa using homozygous knockout mice have clearly established the antioxidant role of Hmox-1 against the toxicity resulting from treatment with endotoxin, heme, and hydrogen peroxide. We rationalized that a modest decrease in Hmox-1 gene expression in Hmox-1+/- hearts might also cripple the cellular antioxidant defense mechanism, which is protective in certain disease models whose pathogenesis is known to be associated with an overproduction of ROS. Similar to what was observed by Poss and Tonegawa,18 the Hmox-1+/- mice generated in our laboratory showed no increased susceptibility to endotoxin toxicity compared with that of wild-type mice (unpublished data). This report documents that heterozygous Hmox-1+/- mice with an inactivated Hmox-1 allele are more vulnerable to myocardial ischemia/reperfusion injury.
Heme oxygenase, which catalyzes the initial rate-limiting step of heme catabolism, is known to be induced by a wide variety of oxidative stresses, including H2O2, UV radiation, and reduced glutathione depletion.3 4 A study from our laboratory documented that ischemia/reperfusion could induce the expression of Hmox-1 mRNA in the heart.8 Induction of Hmox-1 gene expression increased as a function of duration of reperfusion, which could be inhibited with ROS scavengers and SOD, suggesting that oxidative stress developed during ischemia/reperfusion induces the HO-1 gene. Our results are consistent with these previous reports, indicating that Hmox-1 is induced by oxidative stress.
The role of HO-1 in oxidative stress adaptation was further supported by the observation that the hearts of the Hmox-1+/- mice could not be preconditioned to make them resistant to subsequent oxidative stress compared with wild-type hearts. Preconditioning by repeated brief episodes of ischemia/reperfusion is known to develop oxidative stress.19 The development of oxidative stress presumably leads to adaptive modification of the hearts, which is achieved through a chain of reactions including generation of intracellular mediators and modification of signal transduction processes and gene expression.20 Oxidative stress has been shown to function as a second messenger in this process.21 The results of our study suggest that induction of HO-1 by oxidative stress plays a crucial role in preconditioning-mediated adaptive modification of the heart.
HO, an oxidative stress–inducible protein, is also an HSP, HSP 32. HO-1 is evolutionarily conserved in the primary amino acid and nucleotide sequences, and its extent of homology between mouse, rat, and human is >80%. Hmox-1 is readily induced not only by oxidative stress but also with many other pathological and environmental stresses, such as hypoxia, hyperoxia, cellular transformations, and heat shock.22 This enzyme catalyzes the conversion of heme into biliverdin and bilirubin, which function as in vivo antioxidants.1 In addition, a messenger molecule, carbon monoxide (CO), is generated during the production of biliverdin. Previous studies demonstrated that production of cGMP from CO signaling is beneficial for the heart.23
To examine whether antioxidants could rescue the Hmox-1+/- hearts from increased ischemia/reperfusion injury, the Hmox-1+/- mice were treated with 2 different antioxidants, Trolox and NAC. Both of these antioxidants are cell-permeable. Trolox, an analogue of vitamin E, is a hydrophilic antioxidant that has been found to be cardioprotective. NAC is a precursor of glutathione and functions as an antioxidant. NAC also prevents intracellular oxidation and maintains glutathione levels by reducing cystine to cysteine. Both Trolox and NAC provided similar degrees of cardioprotection when used in vitro or in vivo. Neither of these antioxidants, however, could completely rescue the hearts from the ischemia/reperfusion injury. Nevertheless, they significantly reduced the amount of ischemia/reperfusion injury compared with control Hmox-1+/- hearts. Interestingly, the amount of cardioprotection afforded by these antioxidants was comparable to that obtained by preconditioning the Hmox-1+/- hearts.
Numerous reports in the literature indicate
cardioprotective effects of
preconditioning.24 25
Preconditioning potentiates a signal transduction mechanism coupled
with both G-protein and tyrosine kinase receptors, leading to the
activation of protein kinase C and mitogen-activated protein
kinases and redox-sensitive transcription factors such as nuclear
factor-
B and activating
protein-1.26
Preconditioning-mediated cardioprotection is believed to be achieved
through its ability to induce several cardioprotective genes and
proteins, including HSP 27, HSP 70, HSP 89, SOD, and reduced
glutathione peroxidase-1.27
These genes/proteins have been shown to constitute the defense system
of the myocardium. Our results demonstrate that HO-1 is
also an important member of the antioxidant defense system of the
heart.
In summary, our study documents the crucial role of Hmox-1 in cardioprotection. A decrease in HO-1 in the heart makes it vulnerable to ischemia/reperfusion injury. In vivo or in vitro antioxidant therapy only partially rescued the Hmox-1+/- hearts from the ischemia/reperfusion injury, suggesting that HO-1 plays an important role in cardioprotection. Preconditioning, which provokes oxidative stress and induces a large variety of oxidative stress–inducible genes, was unable to rescue the hearts of heterozygous Hmox-1+/- mice from cellular injury, presumably because of the reduced level of Hmox-1. This paves the way for the potential of Hmox-1 gene therapy for cardioprotection in pathological conditions in which oxidative stress is implicated.
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Acknowledgments |
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This study was supported by NIH grants HL-22559, HL-33889, and HL-34360 (Dr Das), a Grant-in-Aid from the American Heart Association and HL-56803 (Dr Maulik), HL-56421 and P30-ES-06639 (Dr Ho), and DK-43135 (Dr Alam).
Received August 25, 2000; revision received October 9, 2000; accepted October 10, 2000.
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References |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Tenhunen R, Marver HS, Schmid R. The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase. Proc Natl Acad Sci U S A. 1968;61:748–755.
2. Maines MD. Heme oxygenase: function, multiplicity, regulatory mechanisms, and clinical applications. FASEB J. 1988;2:2557–2568.[Abstract]
3.
Vile GF,
Tyrrell RM. Oxidative stress resulting from ultraviolet A irradiation
of human skin fibroblasts leads to a heme
oxygenase-dependent increase in ferritin.
J Biol Chem. 1993;268:14678–14681.
4. Tomaro ML, Frydman J, Frydman RB. Heme oxygenase induction by CoCl2, CO-protoporphyrin IX, phenyl hydrazine and diamide: evidence for oxidative stress involvement. Arch Biochem Biophys. 1991;286:610–617.[Medline] [Order article via Infotrieve]
5. Nath KA, Balla G, Vercellotti G. Induction of heme oxygenase is a rapid, protective response in rhabdomyolysis in the rat. J Clin Invest. 1992;90:267–270.
6. Vogt BA, Alam J, Croatt AJ, et al. Acquired resistance to acute oxidative stress: possible role of heme oxygenase and ferritin. Lab Invest. 1995;72:474–483.[Medline] [Order article via Infotrieve]
7.
Vile GF,
Basu-Modak S, Waltner C, et al. Heme oxygenase 1 mediates
an adaptive response to oxidative stress in human skin fibroblasts.
Proc Natl Acad Sci
U S A. 1994;91:2607–2610.
8. Maulik N, Sharma HS, Das DK. Induction of the haem oxygenase gene expression during the reperfusion of ischemic rat myocardium. J Mol Cell Cardiol. 1996;28:1261–1270.[Medline] [Order article via Infotrieve]
9.
Alam J, Cai
J, Smith A. Isolation and characterization of the mouse heme
oxygenase-1 gene. J Biol
Chem. 1994;269:1001–1009.
10. Tybulewicz VLJ, Crawford CE, Jackson PK, et al. Neonatal lethality and lymphopenia in mice with a homozygous disruption of the c-abl proto-oncogene. Cell. 1991;65:1153–1163.[Medline] [Order article via Infotrieve]
11.
Nagy A,
Rossant J, Nagy R, et al. Derivation of completely cell culture-derived
mice from early-passage embryonic stem cells.
Proc Natl Acad Sci
U S A. 1993;90:8424–8428.
12. Yoshida T, Maulik N, Engelman RM, et al. Glutathione peroxidase knockout mice are susceptible to myocardial ischemia reperfusion injury. Circulation. 1997;96(suppl II):II-216–II-220.
13.
Miller DL,
Van Winkle DM. Ischemic preconditioning limits infarct size
following regional ischemia-reperfusion in in situ mouse
hearts. Cardiovasc Res. 1999;42:680–684.
14.
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.
15. Cordis GA, Maulik N, Das DK. Detection of oxidative stress in heart by estimating the dinitrophenylhydrazine derivative of malonaldehyde. J Mol Cell Cardiol. 1995;27:1645–1653.[Medline] [Order article via Infotrieve]
16.
Yoshida T,
Maulik N, Engelman RM, et al. Targeted disruption of the mouse
SodI gene makes the hearts
vulnerable to ischemic reperfusion injury.
Circ Res. 2000;86:264–269.
17.
Poss KD,
Tonegawa S. Heme oxygenase 1 is required for mammalian iron
reutilization. Proc Natl Acad Sci
U S A. 1997;94:10919–10924.
18.
Poss KD,
Tonegawa S. Reduced stress defense in heme oxygenase
1-deficient cells. Proc Natl Acad Sci
U S A. 1997;94:10925–10930.
19. Das DK, Engelman RM, Maulik N. Oxygen free radical signaling in ischemic preconditioning. Ann N Y Acad Sci. 1999;874:49–68.[Medline] [Order article via Infotrieve]
20. Maulik N, Das DK. Redox regulation of ischemic adaptation. In: Sen CK, Sies H, Baeuerle PA, eds. Antioxidant and Redox Regulation of Genes. New York, NY: Academic Press; 1999:491–516.
21. Das DK, Maulik N, Sato M, et al. Reactive oxygen species function as second messengers during ischemic preconditioning of heart. Mol Cell Biochem. 1999;196:59–67.[Medline] [Order article via Infotrieve]
22. Maines MD. The heme oxygenase system: a regulator of second messenger gases. Annu Rev Pharmacol Toxicol. 1997;37:517–554.[Medline] [Order article via Infotrieve]
23. Maulik N, Engelman D, Watanabe M, et al. Nitric oxide/carbon monoxide: a free radical-dependent molecular switch for myocardial preservation during ischemic arrest. Circulation. 1996;94(suppl II):II-398–II-406.
24. Flack JE, Kimura Y, Engelman RM, et al. Preconditioning the heart by repeated stunning improves myocardial salvage. Circulation. 1991;84(suppl III):III-369–III-374.
25. Kukreja RC. Role of KATP channel in heat shock and pharmacological preconditioning. Ann N Y Acad Sci. 1999;874:211–221.[Medline] [Order article via Infotrieve]
26. Maulik N, Yoshida T, Zu Y-L, et al. Ischemic adaptation of heart triggers a tyrosine kinase regulated signaling pathway: a potential role for MAPKAP kinase 2. Am J Physiol. 1998;275:H1857–H1864.
27. Das DK, Maulik N, Moraru II. Gene expression in acute myocardial stress. J Mol Cell Cardiol. 1995;27:181–193. [Medline] [Order article via Infotrieve]
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 D. Frank, C. Kuhn, B. Brors, C. Hanselmann, M. Ludde, H. A. Katus, and N. Frey Gene Expression Pattern in Biomechanically Stretched Cardiomyocytes: Evidence for a Stretch-Specific Gene Program Hypertension, February 1, 2008; 51(2): 309 - 318. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. D. Orozco, M. H. Kapturczak, B. Barajas, X. Wang, M. M. Weinstein, J. Wong, J. Deshane, S. Bolisetty, Z. Shaposhnik, D. M. Shih, et al. Heme Oxygenase-1 Expression in Macrophages Plays a Beneficial Role in Atherosclerosis Circ. Res., June 22, 2007; 100(12): 1703 - 1711. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Zhu, Y. Zhang, B. A. Ojwang, M. A. Brantley Jr, and J. M. Gidday Long-Term Tolerance to Retinal Ischemia by Repetitive Hypoxic Preconditioning: Role of HIF-1{alpha} and Heme Oxygenase-1 Invest. Ophthalmol. Vis. Sci., April 1, 2007; 48(4): 1735 - 1743. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-i. Tsuchihashi, M. Livhits, Y. Zhai, R. W. Busuttil, J. A. Araujo, and J. W. Kupiec-Weglinski Basal Rather Than Induced Heme Oxygenase-1 Levels Are Crucial in the Antioxidant Cytoprotection J. Immunol., October 1, 2006; 177(7): 4749 - 4757. [Abstract] [Full Text] [PDF]
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|
 |
|
 A. S. Pachori, L. G. Melo, L. Zhang, S. D. Solomon, and V. J. Dzau Chronic Recurrent Myocardial Ischemic Injury Is Significantly Attenuated by Pre-Emptive Adeno-Associated Virus Heme Oxygenase-1 Gene Delivery J. Am. Coll. Cardiol., February 7, 2006; 47(3): 635 - 643. [Abstract] [Full Text] [PDF]
|
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 |
|
 K. Kamada, F. S. Gaskin, T. Yamaguchi, P. Carter, T. Yoshikawa, M. Yusof, and R. J. Korthuis Role of calcitonin gene-related peptide in the postischemic anti-inflammatory effects of antecedent ethanol ingestion Am J Physiol Heart Circ Physiol, February 1, 2006; 290(2): H531 - H537. [Abstract] [Full Text] [PDF]
|
|
|
|
|
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L. Wu and R. Wang Carbon Monoxide: Endogenous Production, Physiological Functions, and Pharmacological Applications Pharmacol. Rev., December 1, 2005; 57(4): 585 - 630. [Abstract] [Full Text] [PDF]
|
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|
 |
|
 X. Liu, J. Wei, D. H. Peng, M. D. Layne, and S.-F. Yet Absence of Heme Oxygenase-1 Exacerbates Myocardial Ischemia/Reperfusion Injury in Diabetic Mice Diabetes, March 1, 2005; 54(3): 778 - 784. [Abstract] [Full Text] [PDF]
|
|
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|
|
|
J. Tongers, B. Fiedler, D. Konig, T. Kempf, G. Klein, J. Heineke, T. Kraft, S. Gambaryan, S. M Lohmann, H. Drexler, et al. Heme oxygenase-1 inhibition of MAP kinases, calcineurin/NFAT signaling, and hypertrophy in cardiac myocytes Cardiovasc Res, August 15, 2004; 63(3): 545 - 552. [Abstract] [Full Text] [PDF]
|
|
|
|
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|
 C.-M. Hu, Y.-H. Chen, M.-T. Chiang, and L.-Y. Chau Heme Oxygenase-1 Inhibits Angiotensin II-Induced Cardiac Hypertrophy In Vitro and In Vivo Circulation, July 20, 2004; 110(3): 309 - 316. [Abstract] [Full Text] [PDF]
|
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|
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M. Jain, L. Cui, D. A. Brenner, B. Wang, D. E. Handy, J. A. Leopold, J. Loscalzo, C. S. Apstein, and R. Liao Increased Myocardial Dysfunction After Ischemia-Reperfusion in Mice Lacking Glucose-6-Phosphate Dehydrogenase Circulation, February 24, 2004; 109(7): 898 - 903. [Abstract] [Full Text] [PDF]
|
|
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|
|
F. Eefting, B. Rensing, J. Wigman, W. J. Pannekoek, W. M. Liu, M. J. Cramer, D. J Lips, and P. A Doevendans Role of apoptosis in reperfusion injury Cardiovasc Res, February 15, 2004; 61(3): 414 - 426. [Abstract] [Full Text] [PDF]
|
|
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|
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N. C Chi and J. S Karliner Molecular determinants of responses to myocardial ischemia/reperfusion injury: focus on hypoxia-inducible and heat shock factors Cardiovasc Res, February 15, 2004; 61(3): 437 - 447. [Abstract] [Full Text] [PDF]
|
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G. Yamaura, T. Turoczi, F. Yamamoto, M. A. Q. Siddqui, N. Maulik, and D. K. Das STAT signaling in ischemic heart: a role of STAT5A in ischemic preconditioning Am J Physiol Heart Circ Physiol, July 11, 2003; 285(2): H476 - H482. [Abstract] [Full Text] [PDF]
|
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 |
|
 M. de Perrot, M. Liu, T. K. Waddell, and S. Keshavjee Ischemia-Reperfusion-induced Lung Injury Am. J. Respir. Crit. Care Med., February 15, 2003; 167(4): 490 - 511. [Abstract] [Full Text] [PDF]
|
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S. R. Vulapalli, Z. Chen, B. H. L. Chua, T. Wang, and C.-S. Liang Cardioselective overexpression of HO-1 prevents I/R-induced cardiac dysfunction and apoptosis Am J Physiol Heart Circ Physiol, August 1, 2002; 283(2): H688 - H694. [Abstract] [Full Text] [PDF]
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G. Imamura, A. A. Bertelli, A. Bertelli, H. Otani, N. Maulik, and D. K. Das Pharmacological preconditioning with resveratrol: an insight with iNOS knockout mice Am J Physiol Heart Circ Physiol, June 1, 2002; 282(6): H1996 - H2003. [Abstract] [Full Text] [PDF]
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L. G. Melo, R. Agrawal, L. Zhang, M. Rezvani, A. A. Mangi, A. Ehsan, D. P. Griese, G. Dell'Acqua, M. J. Mann, J. Oyama, et al. Gene Therapy Strategy for Long-Term Myocardial Protection Using Adeno-Associated Virus-Mediated Delivery of Heme Oxygenase Gene Circulation, February 5, 2002; 105(5): 602 - 607. [Abstract] [Full Text] [PDF]
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