ENDOTHELIAL INJURY IN TRAUMA AND ISCHEMIA-REPERFUSION

这个项目的主要目标是描述 创伤后内皮细胞损伤的过程、严重程度和来源 或冠状动脉和内脏血管的缺血+再灌注。 表征将采取生理、体液和 内皮完整性的形态学评估。 生理 评估将侧重于EDRF依赖性和EDRF非依赖性血管扩张剂 体液评价将与确定 类二十烷酸产生（例如，前列环素释放）;和形态学 评估将集中在透射和扫描电子 内皮细胞的显微镜检查。 三个实验设置 待研究的潜在内皮损伤是（a）心肌缺血+ 再灌注，（B）内脏缺血+再灌注，和（c）创伤性 冲击. 体外结果（例如，离体血管环制备物 灌注的器官）将与在本领域中进行的体内研究相关联。 完整的动物 将特别注意准确的时间 所有模型再灌注后内皮损伤的过程。 此外，将进行研究，以确定氧气的作用 衍生的自由基（例如，超氧化物、氢氧根离子）， 自由基清除剂，以及其他介质的作用， 内皮损伤包括细胞因子（例如，TNF α）和粘合剂 蛋白质和分析随后的脂质过氧化反应所造成的 氧衍生自由基的作用。 适当的封闭剂 包括抗超氧离子的人超氧化物歧化酶（h-SOD） (e.g., N-2-巯基丙酰基甘氨酸），对抗氢氧根离子， 转化生长因子（TGF-β）对抗细胞因子，特别是 TNF α和针对整合素表达的单克隆抗体 针对LFA-1的共同亚基（CD/18）的RI5.7 粘附蛋白的异二聚体家族，作用于嗜中性粒细胞和 内皮粘附蛋白。 此外，我们将研究 通过研究灌注的中性粒细胞与内皮细胞的相互作用 心脏和离体冠状动脉和肠系膜动脉环， 和中性粒细胞的缺乏，并通过测量髓过氧化物酶，一个特定的 中性粒细胞标志物在经受缺血的完整动物中， 再灌注 这些研究结果应提供宝贵的基本信息， 心肌缺血、肠缺血和创伤的病理生理学 休克，并应开辟新的治疗方法，这些潜在的 致命的循环系统紊乱

The primary objective of this project is to characterize the time course, severity and source of endothelial cell Injury following trauma or ischemia + reperfusion of the coronary and splanchnic vasculatures. Characterization will take the form of physiological, humoral and morphological assessment of endothelial integrity. The physiological assessment will focus on EDRF-dependent and EDRF-independent vasodilator profiles; the humoral evaluation will be related to determining eicosanoid production (e.g., prostacyclin release); and the morphological assessment will concentrate on transmission and scanning electron microscopy of endothelial cells. The three experimental settings of potential endothelial injury to be studied are (a) myocardial ischemia + reperfusion, (b) splanchnic ischemia + reperfusion, and (c) traumatic shock. In vitro findings (e.g., vascular ring preparations, isolated perfused organs) will be correlated with in vivo studies performed in the intact animal. Particular attention will be paid to the precise time course of endothelial injury following reperfusion in all models. Moreover, studies will be conducted to determine the role of oxygen derived free radicals (e.g., superoxide, hydroxyl ions) using appropriate free radical scavengers, as well as the role of other mediators of endothelial injury including cytokines (e.g., TNFalpha) and adhesive proteins and analysis of subsequent lipid peroxidation resulting from the action of oxygen derived free radicals. Appropriate blocking agents including human superoxide dismutase (h-SOD) against superoxide ions (e.g., N-2-mercaptoproprionyl glycine), against hydroxyl ions, Transforming Growth Factor (TGF-beta) against cytokines especially TNFalpha and against the expression of integrins, the monoclonal antibody (RI5.7) directed against the common subunit (CD/18) of the LFA-1 heterodimer family of adhesive proteins, acting against neutrophil and endothelial adhesive proteins. Additionally, we will study the interaction of neutrophils with the endothelium by studying perfused hearts and isolated coronary and mesenteric artery rings in the presence and absence of neutrophils and by measuring myeloperoxidase, a specific neutrophil marker in intact animals subjected to ischemia and reperfusion. These findings should provide valuable basic information on the pathophysiology of myocardial ischemia, bowel ischemia, and traumatic shock, and should open up new therapeutic approaches to these potentially lethal circulatory disorders.