Microvascular Injury and Pancreatic Enzyme Products

描述(由申请人提供)：重要的未解决问题之一 生理性休克是理解导致心绞痛形成的机制。 炎性级联反应。休克伴随着细胞的激活。 微循环、白细胞浸润、细胞功能障碍、细胞凋亡与器官 失败了。我们最近获得的证据表明，胰酶是一种 产生体液炎症介质的强大来源 缺血性肠病。胰酶在缺血管腔内的阻断作用 肠道可产生高水平的抗炎保护和 多器官衰竭。我们的结果指出了胰腺的一个重要作用 丝氨酸蛋白酶。根据这一证据，我们假设 肠道中的胰腺消化酶可以越过灌木丛逃逸 缺血时的边界细胞屏障和随后产生的体液炎症 通过消化细胞外基质蛋白和其他细胞因子产生的介质 组件。我们的具体目标是：(1)鉴定特定的胰酶 促进体液微血管激活剂产生的活动 酶阻断缺血肠腔内失血性休克 通过将纯化的胰酶引入小鼠的管腔 肠道；(2)检测胰酶在组织中的定位 并测定肠缺血前后的产生量 小鼠肾间质中炎性介质的产生 肠道；(3)研究炎症发生的分子机制 在外周器官中由胰酶产生激活剂后 肠道；以及(4)提纯和鉴定选定的炎性介质 由胰腺匀浆产生的胰腺酶作为一种无处不在的 炎性介质的来源和纯化的胞外消化液 基质蛋白(包括纤维连接蛋白、玻璃连接蛋白和I型、III型和III型 4)丝氨酸蛋白酶。我们将学习，使用最先进的数字 胰酶的体内显微镜、转运和联合作用 大鼠休克时炎性介质的生化鉴定 模特。便于对炎症介质进行生化鉴定 通过从收获的大鼠和 猪胰腺组织。了解炎症的触发机制 休克将为人类带来新的治疗方式。

DESCRIPTION (provided by applicant): One of the important unresolved issues in physiological shock is understanding the mechanism leading to formation of an inflammatory cascade. Shock is accompanied by cell activation in the microcirculation, leukocyte infiltration, cell dysfunction, apoptosis and organ failure. We recently obtained evidence that pancreatic enzymes serve as a powerful source for generation of humoral inflammatory mediators in the ischemic intestine. Blockade of pancreatic enzymes in the lumen of the ischemic intestine leads to high levels of protection against inflammation and multi-organ failure. Our results point to an important role for pancreatic serine proteinases. In accordance with this evidence we hypothesize that pancreatic digestive enzymes in the intestine can escape across the brush border cell barrier during ischemia and thereafter produce humoral inflammatory mediators by digestion of extracellular matrix proteins and other cellular components. Our Specific Aims are (1) to identify specific pancreatic enzyme activities which contribute to humoral microvascular activator production in hemorrhagic shock by enzyme blockade in the lumen of the ischemic intestine and by introduction of purified pancreatic enzymes into the lumen of the small intestine; (2) to examine the pancreatic enzyme localization in the tissue before and after intestinal ischemia and determine the production of inflammatory mediator production in the interstitial space of the small intestine; (3) to examine molecular mechanisms for initiation of inflammation in a peripheral organ after activator production by pancreatic enzymes in the intestine; and (4) to purify and characterize selected inflammatory mediators produced by pancreatic enzymes from homogenates of pancreas as an ubiquitous source of inflammatory mediators and from digests of purified extracellular matrix proteins (including fibronectin, vitronectin, and collagen I, III and IV) with serine proteinases. We will study, using state of the art digital in-vivo microscopy, transport and action of pancreatic enzymes in combination with biochemical identification of inflammatory mediators in the rat shock model. Biochemical identification of the inflammatory mediators is facilitated by availability of large quantities of starting material from harvested rat and pig pancreatic tissue. Understanding the trigger mechanisms for inflammation in shock will lead to new treatment modalities in man.