Overexpression of TGFbeta1 by adeno-associated virus type-2 vector protects myocardium from ischemia-reperfusion injury.
Transforming growth factor beta(1) (TGFbeta(1)) has been purported to protect tissues from ischemia-reperfusion (I-R) injury. This study was designed to examine if overexpression of TGFbeta(1) using adeno-associated virus type 2 (AAV) protects cardiomyocytes from reoxygenation injury. TGFbeta(1) was overexpressed in cultured HL-1 mouse cardiomyocytes by transfection with AAV/TGFbeta(1)(Latent) or with AAV/TGFbeta(1)(ACT) (active TGFbeta(1)). TGFbeta(1) upregulation reduced cardiomyocyte apoptosis and necrosis induced by 24 h of hypoxia followed by 3 h of reoxygenation concomitant with reduction in reactive oxygen species release, activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and NF-kappaB expression. Transfection with AAV/TGFbeta(1)(ACT) was superior to that with AAV/TGFbeta(1)(Latent). To determine if AAV/TGFbeta(1)(ACT) upregulation in vivo would induce cardioprotection from I-R injury, rat hearts were injected with AAV/TGFbeta(1)(ACT) or phosphate-buffered saline (PBS). Six weeks later, TGFbeta(1)(ACT) was upregulated throughout the myocardium. Following I-R, AAV/TGFbeta(1)(ACT)-overexpressing rats had much smaller infarct size (P<0.01 vs PBS group), which was also related to reduced activation of NADPH oxidase and NF-kappaB, and lower levels of malondialdehyde in I-R tissues. These data demonstrate that overexpression of TGFbeta(1) by AAV can protect cardiac tissues from reperfusion injury, possibly via antioxidant mechanism. These findings suggest potential of TGFbeta(1)(ACT) gene therapy for cardioprotection from I-R injury.
Dandapat
A
A
Division of Cardiovascular Medicine, Department of Cardiology, Gene Therapy Program, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
Hu
C P
CP
Li
D
D
Liu
Y
Y
Chen
H
H
Hermonat
P L
PL
Mehta
J L
JL
D000818:Animals; D017209:Apoptosis; D015415:Biomarkers / Q000032:analysis; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D008297:Male; D008315:Malondialdehyde / Q000032:analysis; D051379:Mice; D015428:Myocardial Reperfusion Injury / Q000378:metabolism* / Q000473:pathology; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism* / Q000473:pathology; D019255:NADPH Oxidases / Q000032:analysis; D016328:NF-kappa B / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D017382:Reactive Oxygen Species / Q000032:analysis; D053773:Transforming Growth Factor beta1 / Q000235:genetics*; D015854:Up-Regulation
D015415:Biomarkers; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D053773:Transforming Growth Factor beta1; D008315:Malondialdehyde; D019255:NADPH Oxidases