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(Circulation. 2001;103:877.)
© 2001 American Heart Association, Inc.
Basic Science Reports |
Gene Transfer of Heat-Shock Protein 70 Reduces Infarct Size In Vivo After Ischemia/Reperfusion in the Rabbit Heart
From the Division of Cardiology, Department of Medicine, Medical College of Virginia, Virginia Commonwealth University, Richmond (J.C.C., M.L.H., R.C.K.); the Department of Cardiology, Kanazawa Medical University, Ishikawa, Japan (S.O.); the Robert-Roessle-Klinik, Max-Delbrueck Center for Molecular Medicine, Berlin, Germany (O.W.); and Baylor College of Medicine, Houston, Tex (M.R.S.).
Correspondence to Rakesh C. Kukreja, PhD, Professor, Division of Cardiology, Medical College of Virginia, Virginia Commonwealth University, 1101 E Marshall St, Richmond, VA 23298. E-mail rakesh{at}hsc.vcu.edu
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Abstract |
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Background—Heat-shock protein 70 (HSP 70) plays a role in myocardial protection. No studies are available, however, to show that direct gene transfer of HSP 70 reduces myocardial infarction in vivo.
Methods and Results—Rabbit hearts were injected with vehicle or Ad.HSP70 at 3 sites (1.5x109 pfu, 50 µL/site) in the left ventricle (LV). Four days later, hearts were removed, and expression of inducible (HSP 70) and constitutive (HSC 70) proteins was measured in the LV and right ventricle (RV). Subsets of 5 to 7 animals in the vehicle-, Ad.lacZ-, and Ad.HSP70-treated groups were subjected to 30 minutes of ischemia and 3 hours of reperfusion. Infarct size was measured by tetrazolium staining. Increased expression of HSP 70 was observed in LV injected with Ad.HSP70 compared with vehicle-treated hearts. HSP 70 was undetectable in RV, the noninjected region of the heart. The expression of HSC 70 remained unchanged in hearts treated with vehicle or Ad.HSP70. Infarct size (% risk area) decreased to 24.5±2.8 in Ad.HSP70-injected hearts compared with 41.9±2.8 and 42.7±2.5 in the vehicle- and Ad.LacZ-treated hearts (P<0.01). The infarct size was not different between the vehicle- and Ad.LacZ-treated hearts (P>0.05). The risk areas (% of LV) were not different among the 3 groups, ie, 50.1±5.2, 47.7±3.5, and 53.3±2.9 in vehicle-, Ad.lacZ-, and Ad.HSP70-treated groups (P>0.05).
Conclusions—Direct gene delivery of HSP 70 in vivo reduces the severity of ischemic injury in the heart.
Key Words: myocardial infarction • ischemia • proteins • genes • viruses
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Introduction |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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The stress response is commonly associated with a rapid overexpression of a family of heat-shock proteins.1 The major heat-shock proteins are a set of highly conserved proteins having molecular masses of 27, 70, 82, and 90 kDa. The most abundant and best-studied subset is the 70-kDa (HSP 70) protein family. This protein serves an important role by associating with nascently formed proteins that have not reached their permanent folding state and prevents their denaturation.2 The mechanism of action of HSP 70 and related proteins still remains largely a matter of speculation. There is now compelling genetic and biochemical evidence that these proteins belong to a family of ATP-dependent "unfoldases" implicated in protein assembly/dissociation of multimeric complexes, translocation, and import/export processes across membranes.3
It is well established that preconditioning with brief episodes of ischemia,4 5 6 as well as other forms of stress, such as heat shock,5 6 7 8 result in increased production of HSP 70 in the heart. The amount of HSP 70 synthesis correlates with the extent of myocardial protection after ischemia/reperfusion injury.5 Experimental evidence supports the proposition that in vitro cardiac myocytes transfected with the gene encoding for HSP 70 are protected from ischemic injury.9 No work has been done, however, to show direct transfer of HSP 70 gene in vivo into the beating heart. In the present study, therefore, we sought to investigate the direct cause-and-effect relationship of HSP 70 overexpression in myocardial protection during prolonged ischemia/reperfusion injury in vivo in the rabbit heart. We used recombinant adenovirus encoding for the inducible form of human HSP 70 (Ad.HSP70) to transfer the gene into the cardiac muscle to show whether the increased expression of HSP 70 results in reduction of infarct size subsequent to ischemia/reperfusion injury.
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Methods |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Animals
Male New Zealand White rabbits (2.8 to 3.3 kg) were used for the studies. The rabbits were supplied by the Prince Rabbitry (Oakhill, WVa) or the Blue and Gray Rabbitry (Unionville Lane, Va). The animals were allowed to readjust to the new housing environment for
1
week before the experiment. The care and use of the animals were
conducted in accordance with the guidelines of the Committee on Animals
of Virginia Commonwealth University.
Generation of the Adenoviral Construct
The E1 region of the replication-defective adenoviral
vector was replaced by an expression cassette containing the entire
coding region for human heat-shock protein 70 (HSP70) driven by the
human CMV-IE promoter in parallel to the transcriptional direction of
the adenovirus E1 ORF and terminated by the simian virus 40 large tumor
antigen gene. Ad.HSP70 was generated by cloning human HSP 70 from the
plasmid pH2.3 (ATCC 57494, American Type Culture Collection) as
a BamHI-ScaI fragment into the
BamHI-PmeI sites of the adenoviral shuttle plasmid
pAVC3.10 In a subsequent step, Ad.HSP70 was rescued by homologous recombination of pJM1711 and pAVC3.HSP70 in
293 cells as previously described.12 Ad.HSP70 was
propagated in 293 cells, purified by 2 rounds of CsCl density
centrifugation,12 dialyzed against 1500 mL of PBS with 1 mmol/L MgCl2 and 10% glycerol 4 times at 4°C (1 hour each) with a Slide-A-Lyzer cassette (Pierce), and stored at -80°C. Virus concentration was
determined by measuring absorbency at 260
nm,13 and the titer was
estimated by plaque assay on 293
cells.14 The virus titer was
2x1010 plaque-forming units (pfu) per mL,
and the particle-to-plaque ratio was 
80:1. The presence of
replication-competent adenovirus in the plaque-purified Ad.HSP70
preparation was assessed by infecting A549 cells and excluded by the
absence of lysis of the indicator cells 21 days after
infection.
Experiment Protocol
The rabbits were randomly assigned into 1 of the
following 3 groups: group 1, vehicle, hearts injected with saline
alone; group 2, vector control, hearts injected with adenovirus
encoding irrelevant gene, lacZ (Ad.lacZ); and group 3, Ad.HSP70, hearts
injected with adenovirus Ad.HSP70.
Separate hearts from each of groups 1 and 3 were used to evaluate the expression of HSP 70.
In Vivo Gene Injection
The animals were anesthetized with an intramuscular
injection of ketamine HCl 35 mg/kg and xylazine 5 mg/kg. Further
injections of ketamine/xylazine were given as needed throughout the
surgical procedure. The animals were intubated orotracheally and
ventilated on a positive-pressure ventilator. The tidal volume was set
at 15 mL, and the respiratory rate was adjusted to 30 to 40
cycles/min. Ventilator setting and
PO2
were adjusted as needed to maintain the blood gas parameters within the
physiological range. The surgery was carried out under sterile
conditions. A left thoracotomy was performed at the fourth intercostal
space, and the heart was exposed by stripping the pericardium. With a
26-gauge needle, 150 µL of virus (1.5x109
pfu) or 150 µL of sterile saline was injected directly into the
myocardium at 3 sites in the perceived area at risk. After the
injections, the air was expelled from the chest, and the surgical
wounds were sutured closed. The animals were observed during recovery
until fully conscious and then extubated. The animals received
intramuscular doses of analgesia (buprenorphine 0.02 mg/kg) and
antibiotic (penicillin 200 000 U/kg).
Myocardial Infarction Protocol
Four days after the injection of saline or virus, the
animals were reanesthetized and after tracheotomy, artificially
ventilated with room air. The thorax was reopened and the heart exposed
to identify the coronary artery branch. A ligature was then placed
around the left coronary artery, and the artery was occluded by snaring
with a small tube through which the ligature had been passed. After 30
minutes of ischemia, the ligature was released and the heart reperfused
for 3 hours.
Measurement of Infarction
At the end of the infarction protocol, the ligature
around the coronary artery was retightened, and 1 mL of 10% Evans
blue dye was injected as a bolus into the jugular vein until the eyes
turned blue. The animals were euthanized immediately, and the heart was
removed and frozen. The heart was then cut from apex to base into 6 to
8 transverse slices of equal thickness. The area at risk was determined
by negative staining with Evans blue. The slices were then incubated in
1% triphenyltetrazolium chloride solution in isotonic pH 7.4 phosphate
buffer at 37°C for 20 minutes. The slices were subsequently fixed in
10% formalin solution for 6 hours. Red-stained viable tissue was
easily distinguished from the infarcted pale/unstained necrotic tissue.
The areas of infarcted tissue, the risk zone, and the whole left
ventricle (LV) were determined by computer morphometry with Bioquant
imaging software. The area for each region was averaged from slices.
Infarct size was expressed both as a percentage of the total LV and as
a percentage of the ischemic risk area.
Evaluation of Gene Transfer
Gene transfer in the LV and right ventricle (RV) of
hearts treated with vehicle or Ad.HSP70 was evaluated by Western blot
as described previously15
with a mouse monoclonal antibody cross-reacting to the HSP 70 or the
constitutive form of HSP 70 (HSC 70) (Stressgen Biotechnologies
Corp). The secondary antibody was horseradish
peroxidase–conjugated rabbit anti-mouse
IgG.
Statistical Analysis
All measurements are expressed as group mean±SEM.
Changes in hemodynamics and infarct size variables were analyzed by a
1-way repeated-measures ANOVA to determine the effects of time, group,
and time-by-group interaction. If the global tests showed major
interactions, post hoc contrasts between different time points within
the same group or between different groups were performed with a
t test. Statistical differences
with a value of P<0.05 were
considered significant.
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Results |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Expression of HSP 70
In vivo intraventricular injection of Ad.HSP70 resulted in a robust expression of HSP 70 in the LV compared with the hearts injected with the vehicle (Figure 1
). The noninjected area of the heart, ie, the RV,
did not show HSP 70 expression in vehicle- or Ad.HSP70-treated hearts.
Previous studies have shown that treatment of viral vectors has no
effect on expression of HSP 70 in the
myocytes.16 17
The expression of HSC 70 was not altered in LV and RV with the vehicle
and Ad.HSP70 injections.
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Myocardial Infarction
Ad.HSP70-injected hearts demonstrated significant
reduction in infarct size (% risk area: 24.5±2.8) compared with
41.9±2.9 and 42.7±2.5 in the vehicle- and Ad.lacZ-injected hearts,
respectively (P<0.01,
Figure 2A). There were no significant differences in the
infarct size between the vehicle- and Ad.lacZ-injected hearts
(41.9±2.9 versus 42.7±2.5,
P>0.05), suggesting that the
reduced infarct size observed in Ad.HSP70-injected hearts was entirely
due to the expression of HSP 70 but not HSC 70, which was uniformly
expressed in the LV and RV. A similar pattern was observed when infarct
size was expressed as percentage of LV
(Figure 2B). The risk areas (% of LV) were not significantly
different among the groups, ie, 50.1±5.2, 47.7±3.5, and 53.3±2.9 in
the vehicle-, Ad.lacZ-, and Ad.HSP70-treated groups, respectively
(P>0.05,
Figure 2C).
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Hemodynamics
Heart rate, mean arterial blood pressure, and
rate-pressure product are shown in the
Table.
Except for the indicated differences, these parameters were comparable
among the 3 groups at baseline, during occlusion, and during the
reperfusion period. All groups had a similar decline in blood pressure
after coronary occlusion. During reperfusion, the heart rate, mean
arterial pressure, and rate-pressure product decreased gradually,
sometimes significantly, as indicated in all the
groups.
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Gene therapy is a rapidly expanding field with potential applications to every human organ system.18 19 Recently, adenoviruses have been used as efficient vectors for in vivo gene transfer into the myocardium.20 Using this delivery system, we set out to elucidate the direct cause-and-effect relationship of HSP 70 in inducing antinecrotic cardioprotection in the intact rabbit heart. Our results show that direct injection of Ad.HSP70 into the LV muscle caused robust expression of HSP 70 compared with the RV (the noninjected region). No significant increase in the expression of HSP 70 was observed in the vehicle-treated LV, suggesting that the increased expression in the Ad.HSP70-injected hearts was not due to injection-related stress. The expression of HSC 70 did not change significantly in the LV and RV injected with vehicle, Ad.lacZ, or Ad.HSP70. The infarct area was significantly reduced in the ischemic hearts injected with Ad.HSP70. No significant difference in the infarct size was observed between the vehicle- (saline) and Ad.lacZ-injected hearts. Systemic hemodynamics, ie, heart rate, mean arterial pressure, and rate-pressure product during baseline (preischemia), ischemia, and reperfusion, were not significantly different in the hearts treated with saline, Ad.lacZ, or Ad.HSP70. Taken together, our data show that direct delivery of HSP 70 with multiple injections of Ad.HSP70 into the LV muscle overexpressed the protein, which subsequently protected the heart against ischemia/reperfusion injury by reducing necrosis in the injected region of the heart.
A number of gene transfer approaches have been used, including direct injection of naked plasmid DNA, ex vivo genetically engineered transplanted cells, liposome-DNA complexes, and several recombinant and conjugated viruses.21 Replication-defective adenoviruses have been exploited as gene delivery vehicles because of their ability to infect a wide variety of hosts and tissues. In addition, adenoviruses are highly efficient in infecting slowly replicating or nonreplicating cells, particularly myocytes.22 23 Direct injection of the adenovirus vectors into the heart muscle transduced genes in the transmural region of myocardium, which peaked during the first week20 but was completely extinguished at 30 days. Another study showed the expression of the gene to persist for as long as 55 days after injection, although the magnitude of expression was much lower.24 The cardiac myocytes were the target of the adenovirus-mediated gene transfer, as confirmed by histological examination.20 In the present study, we demonstrated increased expression of HSP 70 four days after injection of Ad.HSP70 into the LV, although long-term expression was not investigated. On the basis of the published studies with lacZ expression, it is possible that the duration of HSP 70 expression by our technique may have lasted for extended periods after in vivo injection. Further studies would be necessary to demonstrate the time course of gene expression with this technique and its correlation with ischemic tolerance in vivo.
Our direct gene transfer approach resulted in significant reduction in infarct size, suggesting that HSP 70 could be added into the number of gene products that can potentially be used clinically to reduce ischemic injury. A previous study demonstrated better functional recovery and less leakage of creatine phosphokinase after ischemia in the hearts transfected with HSP 70 gene than in the control or nontreated hearts.25 This study used a combination of intracoronary infusion of hemagglutinating virus of Japan liposome and heart transplantation to transfer the HSP 70 gene, followed by Langendorff perfusion to evaluate the effect of HSP 70 on myocardial protection. The intracoronary infusion technique caused global transfection of the HSP 70 gene in this study. In contrast, our approach is straightforward and involved direct injection of Ad.HSP70 into LV muscle in vivo, which resulted in localized expression of HSP 70.
Previous studies have shown distinct correlations between
the expression of HSP 70 by pathophysiological stressors and cardiac
resistance to ischemia.5
Transgenic mice overexpressing HSP 70 have been shown to be resistant
to ischemic injury.26A In
contrast, other studies have shown that expression of HSP 70 is not
always related to tissue
protection.8 26B
This is because the acquisition of cardiac resistance to ischemia after
ischemic preconditioning or heat shock is a multifactorial phenomenon
that includes several other possible mechanisms besides the induction
of 1 members of the heat-shock protein family. These include the
activation of protein kinase
C,27 28 receptor
tyrosine kinase,29
stress-activated MAPKAP kinase
2,30 nuclear
factor-
B,31 nitric oxide
synthase(s),32 33
antioxidant defense systems such as increased activities of
SOD34 and
catalase,35 opening of
ATP-sensitive potassium
channels,15 36 or
possibly other unknown mechanisms. Some of the above mediators may have
a role in signaling pathways leading to the synthesis of 1 of the
protective proteins, including HSP
70.37 The present study
suggests that overexpression of HSP 70 by in vivo transfer of the gene
has a direct protective effect, which appears to be independent of
other factors.
In conclusion, the present study provides direct evidence of the protective role of HSP 70 during ischemia/reperfusion in vivo. Our gene delivery approach caused overexpression of the protein in the injected region only, ie, the LV. This in vivo gene transfer technique can be applied to study other candidate genes whose products may have regulatory or physiological effects because they affect the function/response of the heart at the cellular and molecular levels. By use of adenovirus gene constructs of the key mediators and effectors of preconditioning, the direct injection technique can be used to determine the cause-and-effect relationship of a number of other agents that play a role in acquisition of ischemic tolerance in vivo.
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Acknowledgments |
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This work was supported in part by grants HL-51045 and HL-59469 from the National Institutes of Health (Dr Kukreja).
Received June 26, 2000; revision received August 22, 2000; accepted August 23, 2000.
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References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Lindquist S, Craig EA. The heat shock proteins. Annu Rev Genet. 1988;22:631–637.[Medline] [Order article via Infotrieve]
2.
Mirault ME,
Goldschmidt-Clermont M, Artavanis-Tsakonas S, et al.
Organization of the multiple genes for the 70,000 dalton heat shock
protein in Drosophila
melanogaster. Proc Natl Acad
Sci
U S A. 1979;76:5254–5258.
3. Rothman JE. Polypeptide chain binding proteins: catalysts of protein folding and related processes in cells. Cell. 1989;59:591–601.[Medline] [Order article via Infotrieve]
4. Knowlton AA, Brecher P, Apstein CS. Rapid expression of heat shock protein in the rabbit after brief cardiac ischemia. J Clin Invest. 1991;87:139–147.
5.
Marber MS, Latchman
DS, Walker JM, et al. Cardiac stress protein elevation 24 hours after
brief ischemia or heat stress is associated with resistance to
myocardial infarction.
Circulation. 1993;88:1264–1272.
6. Qian Y-Z, Bernardo NL, Nayeem MA, et al. Induction of 72 kilodalton heat shock protein does not produce second window of ischemic preconditioning in rat heart. Am J Physiol. 1999;276:H224–H234.
7.
Currie RW, Tanguay
RM, Kingma JG Jr. Heat-shock response and limitation of tissue necrosis
during occlusion/reperfusion in rabbit hearts.
Circulation. 1993;87:963–971.
8. Qian Y-Z, Shipley JS, Levasseur JE, et al. Dissociation of the expression of 72 and 27 kDa heat shock proteins with ischemic tolerance following heat shock in rat heart. J Mol Cell Cardiol. 1998;30:1163–1172.[Medline] [Order article via Infotrieve]
9.
Hutter MW, Sievers
RE, Barbosa V, et al. Heat-shock protein induction in rat hearts:
direct correlation between the amount of heat-shock protein induced and
the degree of myocardial protection.
Circulation. 1994;89:355–360.
10. Lozier JN, Yankaskas JR, Ramsey WJ, et al. Gut epithelial cells as targets for gene therapy of hemophilia. Hum Gene Ther. 1997;8:1481–1490.[Medline] [Order article via Infotrieve]
11. McGrory WJ, Bautista DS, Graham FL. A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5. Virology. 1988;163:614–617.[Medline] [Order article via Infotrieve]
12. Graham FL, Prevec L. Methods for construction of adenovirus vectors. Mol Biotechnol. 1995;3:207–220.[Medline] [Order article via Infotrieve]
13. Mittereder N, March KL, Trapnell BC. Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J Virol. 1996;70:7498–7509.[Abstract]
14.
Graham FL, Smiley
J, Russell WC, et al. Characteristics of a human cell line transformed
by DNA from human adenovirus type 5.
J Gen Virol. 1977;36:59–74.
15. Hoag JB, Qian Y-Z, Nayeem MA, et al. ATP-sensitive potassium channel mediates delayed ischemic protection by heat stress in rabbit heart. Am J Physiol. 1997;42:H861–H868.
16. Mestril R, Giordano FJ, Conde AG, et al. Adenovirus-mediated gene transfer of a heat shock protein 70 (hsp70i) protects against simulated ischemia. J Mol Cell Cardiol. 1996;28:2351–2358.[Medline] [Order article via Infotrieve]
17. Brar BK, Stephanou A, Wagstaff MJ, et al. Heat shock proteins delivered with a virus vector can protect cardiac cells against apoptosis as well as against thermal or hypoxic stress. J Mol Cell Cardiol. 1999;31:135–146.[Medline] [Order article via Infotrieve]
18.
Anderson WF.
Human gene therapy. Science. 1992;256:808–813.
19.
Quinones MJ, Leor
J, Kloner RA, et al. Avoidance of immune response prolongs expression
of genes delivered to the adult rat myocardium by replication-defective
adenovirus. Circulation. 1996;94:1394–1401.
20.
Guzman RJ,
Lemarchand P, Crystal RG, et al. Efficient gene transfer into
myocardium by direct injection of adenovirus vectors.
Circ Res. 1993;73:1202–1207.
21.
Nabel EG. Gene
therapy for cardiovascular disease.
Circulation. 1995;91:541–548.
22. Brody SL, Crystal RG. Adenovirus-mediated in vivo gene transfer. Ann N Y Acad Sci. 1994;716:90–101.[Medline] [Order article via Infotrieve]
23. Berker KL. Development of adenovirus vectors for the expression of heterologous genes. Biotechniques. 1988;6:616–629.[Medline] [Order article via Infotrieve]
24.
Kass-Eisler A,
Falck PE, Alvira M, et al. Quantitative determination of
adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and
in vivo. Proc Natl Acad Sci
U S A. 1993;90:11498–11502.
25. Suzuki K, Sawa Y, Kaneda Y, et al. In vivo gene transfection with heat shock protein 70 enhances myocardial tolerance to ischemia-reperfusion injury in rat. J Clin Invest. 1997;99:1645–1650.[Medline] [Order article via Infotrieve]
26. Marber MS, Mestril R, Chi SH, et al. Overexpression of the rat inducible 70-KD heat stress protein in a transgenic mouse increases the resistance of the heart to ischemic injury. J Clin Invest. 1995;95:1446–1456.
26. Joannidis M, Cantley LG, Spokes K, et al. Induction of heat-shock proteins does not prevent renal tubular injury following ischemia. Kidney Int. 1995;47:1752–1759.[Medline] [Order article via Infotrieve]
27. Kukreja RC, Qian Y-Z, Okubo S, et al. Role of protein kinase C and 72 kDa heat shock protein in ischemic tolerance following heat stress in the rat heart. Mol Cell Biochem. 1999;195:123–131.[Medline] [Order article via Infotrieve]
28. Baxter GF, Goma FM, Yellon DM. Involvement of PKC in the delayed cytoprotection following sublethal ischemia in rabbit myocardium. Br J Pharmacol. 1995;115:222–224.[Medline] [Order article via Infotrieve]
29. Imagawa J-I, Baxter GF, Yellown DM. Genistein, a tyrosine kinase inhibitor, blocks the "second window of protection" 48 h after ischemic preconditioning in the rabbit. J Mol Cell Cardiol. 1997;29:1885–1893.[Medline] [Order article via Infotrieve]
30. Engel K, Ahlers A, Brach MA, et al. MAPKAP kinase 2 is activated by heat shock and TNF-alpha: in vivo phosphorylation of small heat shock protein results from stimulation of the MAP kinase cascade. J Cell Biochem. 1995;57:321–330.[Medline] [Order article via Infotrieve]
31. Maulik N, Sato M, Price BD, et al. An essential role of NFkappaB in tyrosine kinase signaling of p38 MAP kinase regulation of myocardial adaptation to ischemia. FEBS Lett. 2000;429:365–369.
32. Imagawa J, Yellon DM, Baxter GF. Pharmacological evidence that inducible nitric oxide synthase is a mediator of delayed preconditioning. Br J Pharmacol. 1999;126:701–708.[Medline] [Order article via Infotrieve]
33.
Zhao T, Xi
L, Chelliah J, et al. Inducible nitric oxide synthase mediates delayed
myocardial protection induced by activation of adenosine
A1 receptors: evidence from gene-knockout mice.
Circulation. 2000;102:902–907.
34. 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]
35.
Auyeung Y,
Sievers RE, Weng D, et al. Catalase inhibition with
3-amino-1,2,4-triazole does not abolish infarct size reduction in
heat-shocked rats. Circulation. 1995;92:3318–3322.
36. Bernardo NL, D’Angelo M, Okubo S, et al. Second window of ischemic preconditioning is mediated by opening of ATP-sensitive potassium channels in the rabbit heart. Am J Physiol. 1999;276:H1323–H1330.
37. Okubo S, Bernardo NL, Elliott GT, et al. Tyrosine kinase signaling is involved in the action potential shortening and expression of HSP 72 in late preconditioning. Am J Physiol. 2000;279:H2269–H2276.
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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]
|
|
|
|
 |
|
 L. Grossin, C. Cournil-Henrionnet, A. Pinzano, N. Gaborit, D. Dumas, S. Etienne, J. F. Stoltz, B. Terlain, P. Netter, L. M. Mir, et al. Gene transfer with HSP 70 in rat chondrocytes confers cytoprotection in vitro and during experimental osteoarthritis FASEB J, January 1, 2006; 20(1): 65 - 75. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G.-C. Fan, X. Ren, J. Qian, Q. Yuan, P. Nicolaou, Y. Wang, W. K. Jones, G. Chu, and E. G. Kranias Novel Cardioprotective Role of a Small Heat-Shock Protein, Hsp20, Against Ischemia/Reperfusion Injury Circulation, April 12, 2005; 111(14): 1792 - 1799. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Zhang Prenatal Hypoxia and Cardiac Programming Reproductive Sciences, January 1, 2005; 12(1): 2 - 13. [Abstract] [PDF]
|
|
|
|
|
|
S. Okubo, Y. Tanabe, K. Takeda, M. Kitayama, S. Kanemitsu, R. C. Kukreja, and N. Takekoshi Ischemic preconditioning and morphine attenuate myocardial apoptosis and infarction after ischemia-reperfusion in rabbits: role of {delta}-opioid receptor Am J Physiol Heart Circ Physiol, October 1, 2004; 287(4): H1786 - H1791. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Ren, R. Strube, X. Zhang, S.-Y. Chen, and X. F. Huang Potent Tumor-Specific Immunity Induced by an In vivo Heat Shock Protein-Suicide Gene-Based Tumor Vaccine Cancer Res., September 15, 2004; 64(18): 6645 - 6651. [Abstract] [Full Text] [PDF]
|
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|
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 |
|
 R. Bolli, L. Becker, G. Gross, R. Mentzer Jr, D. Balshaw, and D. A. Lathrop Myocardial Protection at a Crossroads: The Need for Translation Into Clinical Therapy Circ. Res., July 23, 2004; 95(2): 125 - 134. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Li, S. Bae, and L. Zhang Effect of prenatal hypoxia on heat stress-mediated cardioprotection in adult rat heart Am J Physiol Heart Circ Physiol, May 1, 2004; 286(5): H1712 - H1719. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. G. MELO, A. S. PACHORI, D. KONG, M. GNECCHI, K. WANG, R. E. PRATT, and V. J. DZAU Gene and cell-based therapies for heart disease FASEB J, April 1, 2004; 18(6): 648 - 663. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. L. Tang, Y. Tang, Y. C. Zhang, K. Qian, L. Shen, and M. I. Phillips Protection From Ischemic Heart Injury by a Vigilant Heme Oxygenase-1 Plasmid System Hypertension, April 1, 2004; 43(4): 746 - 751. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
 |
|
 J. R. McMullen, T. Shioi, W.-Y. Huang, L. Zhang, O. Tarnavski, E. Bisping, M. Schinke, S. Kong, M. C. Sherwood, J. Brown, et al. The Insulin-like Growth Factor 1 Receptor Induces Physiological Heart Growth via the Phosphoinositide 3-Kinase(p110{alpha}) Pathway J. Biol. Chem., February 6, 2004; 279(6): 4782 - 4793. [Abstract] [Full Text] [PDF]
|
|
|
|
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|
J. A. Yaglom, D. Ekhterae, V. L. Gabai, and M. Y. Sherman Regulation of Necrosis of H9c2 Myogenic Cells upon Transient Energy Deprivation: RAPID DEENERGIZATION OF MITOCHONDRIA PRECEDES NECROSIS AND IS CONTROLLED BY REACTIVE OXYGEN SPECIES, STRESS KINASE JNK, HSP72, AND ARC J. Biol. Chem., December 12, 2003; 278(50): 50483 - 50496. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Rafiee, Y. Shi, K. A. Pritchard Jr., H. Ogawa, A. L. W. Eis, R. A. Komorowski, C. M. Fitzpatrick, J. S. Tweddell, S. B. Litwin, K. Mussatto, et al. Cellular Redistribution of Inducible Hsp70 Protein in the Human and Rabbit Heart in Response to the Stress of Chronic Hypoxia: ROLE OF PROTEIN KINASES J. Biol. Chem., October 31, 2003; 278(44): 43636 - 43644. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Z. Simkhovich, P. Marjoram, C. Poizat, L. Kedes, and R. A. Kloner Brief episode of ischemia activates protective genetic program in rat heart: a gene chip study Cardiovasc Res, August 1, 2003; 59(2): 450 - 459. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. R. Hampton, A. Shimamoto, C. L. Rothnie, J. Griscavage-Ennis, A. Chong, D. J. Dix, E. D. Verrier, and T. H. Pohlman HSP70.1 and -70.3 are required for late-phase protection induced by ischemic preconditioning of mouse hearts Am J Physiol Heart Circ Physiol, July 11, 2003; 285(2): H866 - H874. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. P. Cappola, L. Cope, A. Cernetich, L. A. Barouch, K. Minhas, R. A. Irizarry, G. Parmigiani, S. Durrani, T. Lavoie, E. P. Hoffman, et al. Deficiency of different nitric oxide synthase isoforms activates divergent transcriptional programs in cardiac hypertrophy Physiol Genomics, June 24, 2003; 14(1): 25 - 34. [Abstract] [Full Text] [PDF]
|
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|
|
M. C. LaPointe, X.-P. Yang, O. A. Carretero, and Q. He Left ventricular targeting of reporter gene expression in vivo by human BNP promoter in an adenoviral vector Am J Physiol Heart Circ Physiol, October 1, 2002; 283(4): H1439 - H1445. [Abstract] [Full Text] [PDF]
|
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|
|
J. Jayakumar, K. Suzuki, I. A. Sammut, R. T. Smolenski, M. Khan, N. Latif, H. Abunasra, B. Murtuza, M. Amrani, and M. H. Yacoub Heat Shock Protein 70 Gene Transfection Protects Mitochondrial and Ventricular Function Against Ischemia-Reperfusion Injury Circulation, September 18, 2001; 104 (2009): I-303 - I-307. [Abstract] [Full Text] [PDF]
|
|
|
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|
|
D. Tekin, L. Xi, T. Zhao, M. I. Tejero-Taldo, S. Atluri, and R. C. Kukreja Mitogen-activated protein kinases mediate heat shock-induced delayed protection in mouse heart Am J Physiol Heart Circ Physiol, August 1, 2001; 281(2): H523 - H532. [Abstract] [Full Text] [PDF]
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S. C. FRANCIS, M. K. RAIZADA, A. A. MANGI, L. G. MELO, V. J. DZAU, P. R. VALE, J. M. ISNER, D. W. LOSORDO, J. CHAO, M. J. KATOVICH, et al. Genetic targeting for cardiovascular therapeutics: are we near the summit or just beginning the climb? Physiol Genomics, December 21, 2001; 7(2): 79 - 94. [Abstract] [Full Text] [PDF]
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