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乙醇引起的胰腺外分泌障碍的分子机制。这项拟议的工作旨在确定含酒精饮食对介导胰腺炎的因素的影响。我们的假设是，酒精饮食单独或与低剂量的八肽胆囊收缩素(CCK-8)输注联合使用会导致参与调节促炎细胞因子和趋化因子表达的转录因子激活。这些细胞因子/趋化因子反过来介导炎症和细胞死亡反应，这些反应是胰腺炎的特征。采用大鼠连续灌胃含乙醇饲料的模型，我们提出了以下的具体目标：1.确定酒精饲料加和不加CCK-8对转录因子(即核因子-kappaB和P-1)激活的影响，这些转录因子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.探讨了在有无CCK-8输注的情况下，乙醇饮食诱导胰腺组织中NF-kappaB和AP-1活化的机制(S)。3.观察乙醇饮食加CCK-8和不加CCK-8对胰腺炎大鼠体内特异性细胞因子/趋化因子表达和调控的影响及其在炎症反应和细胞死亡反应中的作用。4.探讨乙醇饮食加和不加CCK-8对胰酶活性的影响及其作用机制(S)。尺寸。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：胰腺胰酶激活；使用凝胶位移分析激活转录因子；以及使用RT-PCR表达细胞因子/趋化因子。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.