Inflammation of reperfused heart: gene deletion

项目二：再灌注小鼠心脏的炎症：基因缺失项目2的主题仍然是心肌缺血后调节心脏白细胞运输的细胞和分子机制的定义。我们和其他人对缺乏细胞粘附分子的小鼠的开发和表征清楚地表明，调节白细胞运输的分子机制是刺激和组织特异性的。最近在使用CAM缺陷小鼠的缺血/再灌注小鼠模型中的研究表明，可利用替代途径进行白细胞外渗。在本申请中，我们将继续追求分子遗传学方法，但重点已经从研究白细胞在缺血/再灌注后增强心肌细胞损伤中的作用转移到了解缺血性损伤后控制心脏白细胞浸润的独特分子机制和细胞粘附分子在缺血性损伤后心脏愈合中的作用。提出了以下具体目标：1）使用细胞粘附分子沿着缺陷的小鼠以及阻断性单克隆抗体，定义在缺血损伤后有和没有再灌注的情况下调节白细胞运输的心脏特异性分子机制。2)通过检查小鼠心脏微血管内皮细胞和来自下腔静脉的小鼠内皮细胞之间的表型和功能差异来表征组织特异性的潜在机制，并通过全面分析细胞因子，趋化因子，在最近开发的小鼠缺血和再灌注的慢性闭胸模型中，在有和没有再灌注的缺血损伤后心脏中的它们的受体; 3）表征和鉴定白细胞与内皮细胞的初级粘附的新途径的分子机制，该途径由IL-4诱导并且不依赖于E-、P-和L-选择素;四、通过在心肌缺血小鼠模型中使用细胞粘附分子缺陷的小鼠来确定细胞粘附分子在调节缺血性损伤后心脏修复和重塑中的作用梗死伴慢性存活。

PROJECT 2: Inflammation of Reperfused Mouse Heart: Gene Deletion The theme of Project 2 continues to be the definition of the cellular and molecular mechanisms that regulate leukocyte trafficking in the heart after myocardial ischemia. Development and characterization of mice deficient in cell adhesion molecules by ourselves and others have clearly demonstrated that the molecular mechanisms that regulate leukocyte trafficking are both stimulus and tissue specific. Recent studies in the mouse model of ischemia/reperfusion using mice deficient in CAMs have shown that alternative pathways can be utilized for leukocyte extravasation. In this application we will continue to pursue a molecular genetic approach, but the focus has shifted away from studying the role of leukocytes in potentiating injury to myocytes after ischemia/reperfusion to understanding the unique molecular mechanisms that control leukocyte infiltration in the heart after ischemic injury and the role of cell adhesion molecules in healing of the heart after ischemic injury. The following specific aims are proposed: 1) define cardiac-specific molecular mechanisms that regulate leukocyte trafficking with and without reperfusion after ischemic injury using mice deficient in cell adhesion molecules along with blocking monoclonal antibodies. 2) characterize potential mechanisms for tissue specificity by examining phenotyping and functional differences between murine cardiac microvascular endothelial cells and murine endothelial cells from the inferior vena cava, and explore potential mechanisms for stimulus specificity by comprehensively profiling mRNA levels for cytokines, chemokines, and their receptors in the heart after ischemic injury with and without reperfusion in a recently developed chronic closed chest model of murine ischemia and reperfusion; 3) characterization and identification of the molecular mechanisms for a novel pathway for the primary adhesion of leukocytes to endothelial cells that is induced by IL-4 and is independent of E-, P-, and L-selectin; 4) definition of the role of cell adhesion molecules in regulating repair and remodeling of the heart after ischemic injury by using mice deficient in cell adhesion molecules in the mouse model of myocardial infarction with chronic survival.