NEURAL REGULATION OF PANCREATIC FUNCTION

描述(改编自申请者摘要)：约10% 急性胰腺炎患者死于胰腺失控 导致大量体液流失和休克的炎症。这项规定 胰腺炎症的发病机制还知之甚少。在气管里，感官 神经通过释放快速激肽来调节炎症 血管内皮细胞和诱导小动脉血管扩张、血浆外渗 中性粒细胞渗入。这种被充分描述的现象被称为 神经源性炎症。这项提议的一般假设是 神经源性机制在急性胰腺炎的发病机制中起重要作用。 胰腺炎。具体地说，我们假设a)速激肽诱导 胰腺的血浆外渗和中性粒细胞浸润 与神经激肽受体相互作用，b)促炎作用 速激肽由细胞表面肽酶终止，c)速激肽 它们的受体在一种广泛使用的急性心肌梗死模型中调节炎症 胰腺炎。由于最近出现了“基因敲除”小鼠， 编码神经激肽受体或细胞表面多肽酶的基因一直是 被同源重组删除，这些实验将在 老鼠。胰腺炎症将通过1)量化和 用伊文思蓝和莫纳星蓝定位血浆外渗， 分别通过以下方式识别和测量内皮细胞间隙 使用银染和光学显微镜观察血浆渗出的情况，3) 用髓过氧化物酶定量和定位中性粒细胞的渗透， 和4)确定水肿的程度和细胞质空泡化 组织学标准。具体目标1将确定以下贡献 外源性和内源性速激肽在急性胰腺发病中的作用 发炎。将确定时间进程和剂量反应，并 介导这些效应的神经激动素受体将通过 拮抗剂和基因敲除小鼠，并使用受体特异性定位 抗血清。《特定目标2》将研究多肽酶在 终止速激肽引起的胰腺炎症。多肽酶 中性内肽酶和血管紧张素转换酶将定位在 使用特异性抗血清的胰腺及其在临床中的重要性 速激肽引起的炎症将使用抑制剂和 基因敲除老鼠。具体目标3将定义感觉神经的重要性， 尤其是速激肽，在急性胰腺炎的发病机制中。 使用神经激肽受体拮抗剂、肽酶抑制剂和基因敲除 小鼠，我们将描绘调节炎症的神经发生机制 急性胰腺炎。

DESCRIPTION (Adapted from Applicant's Abstract): Approximately 10% of patients with acute pancreatitis die from uncontrolled pancreatic inflammation that results in massive fluid losses and shock. The regulation of pancreatic inflammation is poorly understood. In the trachea, sensory nerves regulate inflammation by releasing tachykinins that bind to endothelial cells and induce arteriolar vasodilatation, plasma extravasation and neutrophil infiltration. This well-characterized phenomenon is called neurogenic inflammation. The general hypothesis of this proposal is that neurogenic mechanisms are essential to the pathogenesis of acute pancreatitis. Specifically, we hypothesize that a) tachykinins induce plasma extravasation and neutrophil infiltration in the pancreas by interacting with neurokinin receptors, b) the pro-inflammatory effects of tachykinins are terminated by cell surface peptidases, and c) tachykinins and their receptors regulate inflammation in a widely-used model of acute pancreatitis. Due to the recent availability of "knockout" mice in which the genes encoding neurokinin receptors or cell surface peptidases have been deleted by homologous recombination, these experiments will be performed in mice. Pancreatic inflammation will be assessed by 1) quantifying and localizing plasma extravasation using Evans blue and Monastral blue, respectively, 2) identifying and measuring endothelial cell gaps through which plasma extravasates using a silver stain and light microscopy, 3) quantifying and localizing neutrophil infiltration using myeloperoxidase, and 4) defining the extent of edema, and cytoplasmic vacuolization using histological criteria. Specific Aim 1 will define the contribution of exogenous and endogenous tachykinins to the initiation of acute pancreatic inflammation. The time-course and dose-response will be determined, and the neurokinin receptors that mediate these effects will be identified using antagonists and knockout mice, and localized using receptor-specific antisera. Specific Aim 2 will examine the role of peptidases in the termination of tachykinin-induced pancreatic inflammation. The peptidases neutral endopeptidase and angiotensin converting enzyme will be localized in the pancreas using specific antisera, and their importance in tachykinin-induced inflammation will be determined using inhibitors and knockout mice. Specific Aim 3 will define the importance of sensory nerves, and specifically tachykinins, in the pathogenesis of acute pancreatitis. Using neurokinin receptor antagonists, peptidase inhibitors, and knockout mice, we will delineate the neurogenic mechanisms that regulate inflammation in acute pancreatitis.