ALCOHOL SENSITIZES THE PANCREAS FOR CK INDUCED PANCREATITIS

本应用中描述的工作的广泛长期目标是确定乙醇引起的外分泌胰腺疾病的分子机制。提出的工作旨在确定含乙醇饮食对介导胰腺炎因素的影响。我们的假设表明，单独的乙醇饮食或与低剂量的胆囊收缩素八肽（CCK-8）输注联合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/趋化因子介导炎症和细胞死亡反应，这是胰腺炎的标志。采用连续输注含乙醇饮食的大鼠模型，我们提出了以下具体目的：确定有或没有CCK-8输注的乙醇饮食对转录因子（即NF- κ b和P-1）激活的影响，这些转录因子调节胰腺细胞因子/趋化因子的产生以及炎症和细胞死亡反应。2. 确定在CCK-8输注和不输注的情况下，乙醇饮食引起胰腺NF-kappaB和AP-1活化的机制。3. 确定添加和不添加CCK-8的乙醇饮食对胰腺炎中特定细胞因子/趋化因子表达和调节的影响及其在介导炎症和细胞死亡反应中的作用。4. 确定添加和不添加CCK-8的乙醇饲粮对胰腺内胰蛋白酶激活的影响以及这些影响胰腺内胰蛋白酶激活的机制。测量。为了实现这些目标，将使用组织学技术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、中性粒细胞和巨噬细胞：胰蛋白酶激活；利用凝胶移位法激活转录因子；以及细胞因子/趋化因子的表达。Western blot分析，免疫细胞化学和生物测定。本应用程序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.