NEURAL REGULATION OF PANCREATIC FUNCTION

描述（改编自申请人摘要）：约10% 急性胰腺炎患者死于不受控制的胰腺炎 导致大量体液流失和休克的炎症。 调控 胰腺炎的发病机制知之甚少 在气管中，感觉 神经通过释放与细胞结合的速激肽来调节炎症 内皮细胞并诱导小动脉血管扩张、血浆外渗 和中性粒细胞浸润。 这种特征鲜明的现象被称为 神经源性炎症 这一建议的一般假设是， 神经原性机制在急性脑梗死的发病机制中至关重要， 胰腺炎 具体来说，我们假设a）速激肽诱导 胰腺中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） 速激肽由细胞表面肽酶终止，和c）速激肽 它们的受体在广泛使用的急性炎症模型中调节炎症 胰腺炎 由于最近可用的“敲除”小鼠， 编码神经激肽受体或细胞表面肽酶的基因已经被 通过同源重组删除，这些实验将在 小鼠 胰腺炎症将通过1）定量和 使用伊文思蓝和单星蓝定位血浆外渗， 2）识别和测量内皮细胞间隙， 使用银染色和光学显微镜检查血浆外渗，3） 使用髓过氧化物酶定量和定位嗜中性粒细胞浸润， 以及4）使用细胞内空泡形成来确定水肿和细胞质空泡形成的程度。 组织学标准。 具体目标1将确定以下方面的贡献： 外源性和内源性速激肽对急性胰腺炎的启动作用 炎症 将确定时间过程和剂量反应， 神经激肽受体介导这些影响将确定使用 拮抗剂和敲除小鼠，并使用受体特异性 抗血清 具体目标2将研究肽酶在 终止速激肽诱导的胰腺炎症。 肽酶 中性内肽酶和血管紧张素转换酶将定位于 使用特异性抗血清的胰腺，以及它们在 将使用抑制剂测定速激肽诱导的炎症， 敲除小鼠 具体目标3将定义感觉神经的重要性， 特别是速激肽在急性胰腺炎发病机制中的作用。 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.