ALCOHOL SENSITIZES THE PANCREAS FOR CK INDUCED PANCREATITIS

本申请中描述的工作的广泛的长期目标是确定由乙醇引起的胰腺外分泌紊乱的分子机制。这项工作的目的是确定含乙醇的饮食对胰腺炎介导因素的影响。我们的假设表明，乙醇饮食单独或结合低剂量的胆囊收缩素-八肽（CCK-8）输注导致参与调节促炎细胞因子和趋化因子表达的转录因子的激活。反过来，这些细胞因子/趋化因子介导作为胰腺炎标志的炎症和细胞死亡反应。使用大鼠模型的连续输注含乙醇的饮食，我们提出了以下具体目标：1。确定有和没有CCK-8输注的乙醇饮食对转录因子活化的影响（即，NF-κ B和P-1），其调节胰腺中细胞因子/趋化因子的产生以及炎症和细胞死亡反应。2.确定在存在和不存在CCK-8输注的情况下由乙醇饮食引起的胰腺中NF-κ B和AP-1活化的机制。3.确定有和没有CCK-8输注的乙醇饮食对特定细胞因子/趋化因子的表达和调节的影响及其在胰腺炎中介导炎症和细胞死亡反应的作用。4.确定含和不含CCK-8输注的乙醇饮食对胰腺内胰蛋白酶激活的影响以及这些影响对胰腺内胰蛋白酶激活的机制。测量.实现这些目标的方法包括血清淀粉酶和脂肪酶、胰腺重量、胰腺坏死、细胞凋亡、空泡化、嗜中性粒细胞和巨噬细胞，使用组织学技术：胰蛋白酶活化;使用凝胶迁移分析的转录因子活化;和使用RT-PCR的细胞因子/趋化因子表达。免疫印迹分析、免疫细胞化学和生物测定。本申请中描述的研究结果将阐明乙醇在胰腺炎中的作用机制，可用于设计治疗干预和临床试验的策略。

The broad long-term objective of the work described in this application is to determine the molecular mechanisms responsible for the exocrine pancreatic disorders caused by ethanol. The work proposed is designed to determine the effect of an ethanol-containing diet on the factors that mediate pancreatitis. Our hypothesis states that a ethanol diet alone or in combination with a low dose of cholecystokinin-octapeptide (CCK-8) infusion results in activation of transcription factors involved in regulating the expression of pro-inflammatory cytokines and chemokines. These cytokines/chemokines, in turn, mediate the inflammatory and cell death responses that are the hallmarks of pancreatitis. Using a rat model of continuous infusion of an ethanol-containing diet, we proposed the following specific aims: 1. Determine the effect of the ethanol diet with and without the CCK-8 infusion on activation of transcription factors (i.e., NF- kappaB and P-1) that regulate cytokine/chemokine production in the pancreas and inflammatory and cell death responses. 2. Determine the mechanism(s) of NF-kappaB and AP-1 activation in the pancreas caused by the ethanol diet in the presence and absence of the CCK-8 infusion. 3. Determine the effect of the ethanol diet with and without the CCK-8 infusion on the expression and regulation of specific cytokines/chemokines and their role in mediating inflammatory and cell death responses in pancreatitis. 4. Determine the effect of the ethanol diet with and without the CCK-8 infusion on intrapancreatic trypsin activation and the mechanism(s) of these effects on intrapancreatic trypsin activation. Measurements. to accomplish these goals will include serum amylase and lipase, pancreatic weight, pancreatic necrosis, apoptosis, vacuolization, neutrophils and macrophages using histologic techniques: pancreatic trypsin activation; transcription factor activation using gel shift assays; and expression of cytokines/chemokines using RT-PCR. Western blot analysis, immunocytochemistry and bioassay. The results of the studies described in this application will be elucidation of the mechanism of ethanol's effects in pancreatitis that can be used to design strategies for therapeutic interventions and clinical trials.