Dandapat A, Hu C P, Li D, Liu Y, Chen H, Hermonat P L, Mehta J L
Division of Cardiovascular Medicine, Department of Cardiology, Gene Therapy Program, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
Gene Ther. 2008 Mar;15(6):415-23. doi: 10.1038/sj.gt.3303071. Epub 2007 Nov 15.
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.
转化生长因子β1（TGFβ1）据称可保护组织免受缺血再灌注（I-R）损伤。本研究旨在检验使用2型腺相关病毒（AAV）过表达TGFβ1是否能保护心肌细胞免受复氧损伤。通过用AAV/TGFβ1（潜伏型）或AAV/TGFβ1（活性型）（活性TGFβ1）转染，使培养的HL-1小鼠心肌细胞中TGFβ1过表达。TGFβ1上调减少了缺氧24小时后再复氧3小时诱导的心肌细胞凋亡和坏死，同时伴随着活性氧释放减少、烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶激活和NF-κB表达降低。用AAV/TGFβ1（活性型）转染优于用AAV/TGFβ1（潜伏型）转染。为了确定体内AAV/TGFβ1（活性型）上调是否能诱导对I-R损伤的心脏保护作用，给大鼠心脏注射AAV/TGFβ1（活性型）或磷酸盐缓冲盐水（PBS）。六周后，整个心肌中TGFβ1（活性型）上调。I-R后，过表达AAV/TGFβ1（活性型）的大鼠梗死面积小得多（与PBS组相比，P<0.01），这也与I-R组织中NADPH氧化酶和NF-κB的激活减少以及丙二醛水平降低有关。这些数据表明，AAV介导的TGFβ1过表达可通过抗氧化机制保护心脏组织免受再灌注损伤。这些发现提示了TGFβ1（活性型）基因治疗对I-R损伤心脏保护的潜力。
