PATHOGENESIS OF NEONATAL NECROTIZING ENTEROCOLITIS

本建议的重点是表征新生动物模型 坏死性小肠结肠炎（NEC），并探讨其发病机制 这种毁灭性的疾病的早产儿，因为它涉及到PAF和 其他炎症介质。 虽然成年人的肠道损伤 动物已被证明主要是由PAF介导的， 新生儿NEC可能与成人肠坏死有显著差异， PAF在NEC中的重要性尚不清楚。 新生儿NEC将 在新生大鼠中复制，以模拟人类条件， 动物暴露于NEC的已知风险因素，即配方饲料， 缺氧和细菌定植。 随后，PAF和 其他介质（内毒素、内皮素、肿瘤坏死因子和 血栓烷）将在该模型中通过1）测量血浆（所有） 2）研究PAF对肠组织PAF表达的调控作用， 活性（使用酶测定、Norther印迹分析和原位杂交） 3）研究PAF受体的结合调节和受体 基因转录，和4）抑制PAF和其他介质使用 特异性抑制剂和单克隆抗体。 此外，内皮 细胞单层将用于评价细胞内的相互作用。 缺氧、PAF和内皮素之间的水平。 最后，早产儿 将对新生儿进行研究，以检验喂养刺激 内毒素释放、磷脂酶A2活化、PAF合成和NEC。 这些研究的结果可能会为这些疾病提出新的治疗策略。 预防和治疗新生儿NEC。 候选人是一位受过生物化学训练的学术型生物学家， PAF的评价，因为它与新生儿病理生理学有关。 应用 临床研究者奖旨在支持受保护的时间 远离临床问题，以巩固分子 生物学技术，包括北方印迹分析和原位 杂交方法 随着这些技能的获得，这位研究人员 将独立竞争校外资助。 分子生物学培训将由申办者提供，申办者是 已成立的调查员;这项培训将在儿童 申办者在Memorial医院维护其实验室。 这个奖项 将使候选人能够发展和完成所需的技能 我在埃文斯顿医院的实验室里进行了这些研究。

The emphasis of this proposal is to characterize a neonatal animal model of necroitizing enterocolitis (NEC) and to investigate the pathogenesis of this devastating disease of premature infants as it relates to PAF and other inflammatory mediators. Although intestinal injury in adult animals has been shown to be predominantly mediated by PAF, the etiology of neonatal NEC may be significantly different from adult bowel necrosis, and the importance of PAF in NEC in unknown. Neonatal NEC will be reproduced in newborn rats to simulate the human condition by exposing the animals to the known risk factors of NEC, i.e. formula feeding, hypoxia, and bacterial colonization. Subsequently, the roles of PAF and other mediators (endotoxin, endothelin, tumor necrosis factor, and thromboxane) will be evaluated in this model by 1) measuring plasma (all) and intestinal PAF, 2) studying PAF regulating enzyme expression and activity (using enzyme assays, Norther blot analysis, and in situ hybridization), 3) studying PAF receptor binding regulation and receptor gene transcription, and 4) inhibition of PAF and other mediators using specific inhibitors and monoclonal antibodies. In addition, endothelial cell monolayers will be used to evaluate interactions at the cellular level between hypoxia, PAF, and endothelin. Finally, premature human neonates will be studied to test the hypothesis that feeding stimulates endotoxin release, phospholipase A2 activation, PAF synthesis, and NEC. Results from these studies may suggest new therapeutic strategies for the prevention and treatment of neonatal NEC. The candidate is an academic neonatologist trained in the biochemical evaluation of PAF as it relates to neonatal pathophysiology. Application for the Clinical Investigator Award is designed to support protected time away from clinical concerns in order to solidify training in molecular biological techniques including Northern blot analysis and in situ hybridization. With the acquisition of these skills, this researcher will become competitive independently for extramural funding. Molecular biology training will be provided by the sponsor who is an established investigator; this training will take place at Children's Memorial Hospital where the sponsor maintains her laboratory. This award will enable the candidate to develop and skills necessary to accomplish these pursuits in my laboratory at Evanston Hospital.