ACUTE PANCREATITIS--ROLES OF CYTOKINES AND ANTIOXIDANTS

这些研究的总体目标是应用分子，细胞， 和转基因方法来阐明炎症和 氧化应激在急性胰腺炎小鼠。这些研究将 还提供了有关机制的重要信息， 胰腺在急性胰腺炎期间可以得到保护。的机制 诱发急性胰腺炎的方法有哪些 对疾病的了解很少。因此，这种毁灭性的疾病仍然是一个 严重的临床问题，治疗主要是支持性的。 炎性细胞因子与急性胰腺炎有关，但 细胞因子基因表达的调节尚未被研究和直接 尚未获得关于其作用的证据。其他近期研究 提示自由基可能在急性胰腺炎发病中起重要作用。 氧化应激激活了几种保护基因的表达， 这些抗氧化剂基因的调节尚未被研究， 急性胰腺炎此外，潜在的保护作用 细胞内抗氧化剂在急性胰腺炎中的作用还没有被彻底 考察因此，这项建议的具体目标是： 研究细胞因子在雨蛙素-或 胆碱缺乏的乙硫氨酸补充饮食诱导的胰腺炎 小鼠;和2）检查氧化应激在这些模型中的作用， 急性胰腺炎目标1将通过确定时间 胰腺肿瘤坏死表达的空间模式 因子-α、白细胞介素-1（α和β）和白细胞介素-6基因， 北方印迹、原位杂交和免疫组化。的 重组细胞因子对基因表达和细胞功能的影响 在分离的胰腺腺泡中， 在胰腺中组成型表达细胞因子或细胞因子拮抗剂 将被创建为模型，以描述的职能作用， 急性胰腺炎的发病机制目标2将通过以下方式实现： 确定表达的时间和空间模式，在急性 胰腺炎，抗氧化基因编码金属硫蛋白，血红素 加氧酶和锰超氧化物歧化酶。此外，外源性 抗氧化剂和氧化应激诱导化学物质对激活 基因表达和急性胰腺炎的诱导将被检查 在胰腺中的体内试验和在培养的胰腺腺泡中的体外试验。 最后，细胞内抗氧化剂的功能作用 金属硫蛋白在急性胰腺炎将检查使用遗传小鼠 有针对性地删除金属硫蛋白基因的模型， 在包括胰腺在内的许多组织中过度表达金属硫蛋白，或 在胰腺腺泡细胞中特异性过表达金属硫蛋白。

The overall objective of these studies is to apply molecular, cellular, and transgenic approaches to elucidate the roles of inflammation and oxidative stress in acute pancreatitis In the mouse. These studies will also provide important information regarding mechanisms by which the pancreas can be protected during acute pancreatitis. The mechanisms of induction of acute pancreatitis aryl methods to prevent and treat this disorder are poorly understood. Thus, this devastating illness remains a significant clinical problem, and treatment is largely supportive. Inflammatory cytokines have been implicated in acute pancreatitis, but the regulation of cytokine gene expression has not been examined and direct evidence for their roles has not been obtained. Other recent studies indicate that free radicals may play a key role In acute pancreatitis. Oxidative stress activates the expression of several protective genes, but the regulation of these antioxidant genes has not been examined during acute pancreatitis. Furthermore, the potential protective roles of intracellular antioxidants in acute pancreatitis have not been thoroughly examined. The specific aims of this proposal are, therefore, to: 1) Examine the regulation and potential roles of cytokines in caerulein- or choline-deficient ethionine-supplemented diet-Induced pancreatitis in the mouse; and 2) examine the roles of oxidative stress in these models of acute pancreatitis. Aim 1 will be approached by determining the temporal and spatial patterns of pancreatic expression of the tumor necrosis factor-alpha, Interleukin-1 (alpha and beta) and interleukin-6 genes using Northern blotting, in situ hybridization and immunohistochemistry. The effects of recombinant cytokines on gene expression and cellular functions in isolated pancreatic acini will be examined and transgenic mice that constitutively express cytokines or cytokine antagonists in the pancreas will be created to serve as models for delineating the functional roles of inflammation in acute pancreatitis. Aim 2 will be approached by determining the temporal and spatial patterns of expression, during acute pancreatitis, of the antioxidant genes encoding metallothionein, heme oxygenase and Mn-superoxide dismutase. In addition, effects of exogenous antioxidants and oxidative stress-Inducing chemicals on the activation of gene expression and the induction of acute pancreatitis will be examined in vivo in the pancreas, and in vitro in cultured pancreatic acini. Finally, the functional role of the intracellular antioxidant metallothionein In acute pancreatitis will be examined using genetic mouse models that have targeted deletions of the metallothionein genes, that over-express metallothionein in many tissues including the pancreas, or that over-express metallothionein specifically in pancreatic acinar cells.