SCOR IN ISCHEMIC HEART DISEASE

这个提议的缺血性心脏病SCOR是一个多学科实验室 和临床研究计划，重点是细胞机制， 缺血后心肌炎症，强调的作用 内皮促进早期中性粒细胞（PMN）靶向和可能的PMN- 介导的心肌再灌注损伤。 具体目标是 确定（1）中性粒细胞和内皮细胞之间的粘附作用， 缺血性心肌炎症，发生在内皮细胞的变化， 粘附分子和肌动蛋白细胞骨架组织的细胞表达 介导PMN在再灌注心肌中的靶向和积聚， 和负责的机制，包括当地生产的 促炎细胞因子，内皮细胞胞浆Ca 2+增加 浓度和酪氨酸激酶活性的改变与 受体和粘着斑部位，（2）氧自由基的作用， 缺血后心肌炎症，包括它们在增强 中性粒细胞和内皮细胞粘附分子表面表达和合成 和补体的激活，以及负责 内皮细胞和中性粒细胞中自由基的产生，（3） 内皮源性一氧化氮（NO）在改善缺血后 炎症，通过不同亚型控制NO产生的因素 一氧化氮合酶（NOS）的缺血/再灌注后，和 负责细胞内定位、遗传调节 以及内皮细胞NOS的表达（4）是否抑制PMN治疗 激活和/或粘附到内皮细胞上， 再灌注，可以产生有意义的和永久性的减少梗死 大鼠和犬动物模型中的大小，与改善左 心室（LV）功能和I.V重塑减少，无阴性 对梗死愈合或感染易感性的影响，（5）是否 联合核磁共振成像的局部左心室功能， 射频标签和空间定位的31 P-NMR光谱 心肌高能磷酸盐浓度的测量可以 成功区分功能失调性坏死和存活心肌， 缺血/再灌注后患者。 拟采用的技术 包括分子克隆、逆转录聚合酶链反应 （RT-PCR）、原位RT-PCR、反义核酸酶抑制 寡核苷酸，免疫组织学，免疫电镜，荧光 显微镜，测量细胞内pH和Ca 2+浓度， 荧光指示剂用电子测量氧自由基和NO 顺磁共振，联合核磁共振成像 和空间定位的31 P光谱，以及使用培养的细胞， 灌注心脏和体内动物模型。 我们预计， 拟议的SCOR计划将产生有关细胞的基本信息， 参与缺血后炎症早期阶段的机制， 将提供新的治疗方法，以减少缺血后 心肌损伤

This proposed Ischemic Heart Disease SCOR is a multidisciplinary laboratory and clinical research program focused on the cellular mechanisms underlying post-ischemic myocardial inflammation, emphasizing the role of the endothelium in promoting early neutrophil (PMN) targeting and possible PMN- mediated myocardial reperfusion injury. The specific objectives are to determine (1) the role of adhesion between PMNs and endothelium in post- ischemic myocardial inflammation, the changes that occur in endothelial cell expression of adhesion molecules and actin cytoskeletal organization that mediate targeting and accumulation of PMNs in reperfused myocardium, and the mechanisms responsible, including local production of proinflammatory cytokines, increases in endothelial cell cytoplasmic Ca2+ concentration, and alterations in tyrosine kinase activity associated with receptors and focal adhesion sites, (2) the role of oxygen free radicals in post-ischemic myocardial inflammation, including their role in enhanced surface expression and synthesis of PMN and endothelial adhesion molecules and activation of complement, and the cellular mechanisms responsible for free radical generation in endothelial cells and PMNs, (3) the role of endothelial-derived nitric oxide (NO) in modifying post-ischemic inflammation, the factors controlling NO production by different isoforms of nitric oxide synthase (NOS) following ischemia/reperfusion, and the mechanisms responsible for intracellular localization, genetic regulation and expression of endothelial cell NOS (4) whether therapy to inhibit PMN activation and/or adhesion to endothelial cells, given shortly before reperfusion, can produce a meaningful and permanent reduction in infarct size in rat and dog animal models, associated with improved left ventricular (LV) function and reduced I.V remodelling without a negative impact on infarct healing or susceptibility to infection, (5) whether combined NMR imaging of regional left ventricular function using radiofrequency tags and spatially localized 31P-NMR spectroscopic measurements of myocardial high energy phosphate concentrations can successfully differentiate dysfunctional necrotic and viable myocardium in patients following ischemia/reperfusion. The techniques to be utilized include molecular cloning, reverse transcriptase polymerase chain reaction (RT-PCR), in situ RT-PCR, enzyme inhibition with antisense oligonucleotides, immunohistology, immuno-electron microscopy, fluorescence microscopy, measurement of intracellular pH and Ca2+ concentration with fluorescent indicators, measurement of oxygen radicals and NO by electron paramagnetic resonance (EPR), combined nuclear magnetic resonance imaging and spatially localized 31P spectroscopy, and use of cultured cells, perfused hearts and in vivo animal models. We anticipate that this proposed SCOR program will yield fundamental information about the cellular mechanisms involved in the early stages of post-ischemic inflammation and will provide novel therapeutic approaches to reducing post-ischemic myocardial injury.