PATHOGENESIS OF NEONATAL NECROTIZING ENTEROCOLITIS

这项建议的重点是描述一种新生儿动物模型 坏死性小肠结肠炎(NEC)的发病机制 早产儿的这种破坏性疾病与PAF和 其他炎症介质。尽管成人的肠道损伤 动物已被证明主要是由PAF介导的，即病原学 新生儿NEC的发病率可能与成人肠坏死显著不同， PAF在NEC中的重要性未知。新生儿NEC将是 在新生大鼠身上复制，通过暴露在 动物对已知的NEC危险因素，即配方奶喂养， 低氧和细菌定植。随后，PAF和PAF的作用 其他介质(内毒素、内皮素、肿瘤坏死因子和 血栓烷)在这个模型中将通过1)测量血浆(ALL)来评估 和肠道PAF，2)研究PAF调节酶的表达和 活性(使用酶分析、Norther印迹分析和原位 杂交)，3)研究PAF受体结合调节和受体 基因转录，以及4)抑制PAF和其他介质使用 特定的抑制物和单抗。此外，内皮细胞 细胞单层将用于评估细胞的相互作用 低氧、PAF、ET之间的水平。最后，早产儿 将对新生儿进行研究，以检验喂奶刺激的假设 内毒素释放、磷脂酶A2激活、PAF合成和NEC。 这些研究的结果可能会提出新的治疗策略 新生儿NEC的防治。 应聘者是一位训练有素的新生儿生化学家。 评价PAF与新生儿病理生理的关系。应用 临床研究人员奖旨在支持受保护的时间 远离临床问题，以巩固分子方面的培训 包括Northern印迹分析和原位杂交在内的生物技术 杂交。随着这些技能的获得，这位研究人员 将独立成为外部资金的竞争对手。 分子生物学培训将由赞助商提供， 公认的调查员；这一培训将在儿童 赞助商负责维护其实验室的纪念医院。本奖项 将使应聘者能够发展所需的技能，以完成 这些都是我在埃文斯顿医院的实验室里的追求。

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.