Matherne G P, Linden J, Byford A M, Gauthier N S, Headrick J P
Department of Pediatrics, Cardiovascular Research Center, University of Virginia, Charlottesville, VA 22908, USA.
Proc Natl Acad Sci U S A. 1997 Jun 10;94(12):6541-6. doi: 10.1073/pnas.94.12.6541.
Activation of myocardial A1 adenosine receptors (A1AR) protects the heart from ischemic injury. In this study transgenic mice were created using the cardiac-specific alpha-myosin heavy chain promoter and rat A1AR cDNA. Heart membranes from two transgene positive lines displayed approximately 1,000-fold overexpression of A1AR (6,574 +/- 965 and 10,691 +/- 1,002 fmol per mg of protein vs. 8 +/- 5 fmol per mg of protein in control hearts). Compared with control hearts, transgenic Langendorff-perfused hearts had a significantly lower intrinsic heart rate (248 beats per min vs. 318 beats per min, P < 0. 05), lower developed tension (1.2 g vs. 1.6 g, P < 0.05), and similar coronary resistance. The difference in developed tension was eliminated by pacing. Injury of control hearts during global ischemia, indexed by time-to-ischemic contracture, was accelerated by blocking adenosine receptors with 50 microM 8-(p-sulfophenyl) theophylline but was unaffected by addition of 20 nM N6-cyclopentyladenosine, an A1AR agonist. Thus A1ARs in ischemic myocardium are presumably saturated by endogenous adenosine. Overexpressing myocardial A1ARs increased time-to-ischemic contracture and improved functional recovery during reperfusion. The data indicate that A1AR activation by endogenous adenosine affords protection during ischemia, but that the response is limited by A1AR number in murine myocardium. Overexpression of A1AR affords additional protection. These data support the concept that genetic manipulation of A1AR expression may improve myocardial tolerance to ischemia.
心肌A1腺苷受体(A1AR)的激活可保护心脏免受缺血性损伤。在本研究中,使用心脏特异性α-肌球蛋白重链启动子和大鼠A1AR cDNA创建了转基因小鼠。来自两个转基因阳性品系的心脏膜显示A1AR过表达约1000倍(每毫克蛋白质6574±965和10691±1002 fmol,而对照心脏为每毫克蛋白质8±5 fmol)。与对照心脏相比,转基因Langendorff灌注心脏的固有心率显著降低(每分钟248次搏动 vs. 每分钟318次搏动,P < 0.05),舒张期张力降低(1.2 g vs. 1.6 g,P < 0.05),冠状动脉阻力相似。通过起搏消除了舒张期张力的差异。在整体缺血期间,对照心脏的损伤以缺血性挛缩时间为指标,用50μM 8-(对磺基苯基)茶碱阻断腺苷受体可加速损伤,但添加20 nM N6-环戊基腺苷(一种A1AR激动剂)则无影响。因此,缺血心肌中的A1AR可能被内源性腺苷饱和。过表达心肌A1AR可增加缺血性挛缩时间并改善再灌注期间的功能恢复。数据表明,内源性腺苷激活A1AR在缺血期间提供保护,但该反应受小鼠心肌中A1AR数量的限制。A1AR的过表达提供了额外的保护。这些数据支持这样的概念,即对A1AR表达进行基因操作可能会提高心肌对缺血的耐受性。