LPS-Peptide Interaction in Bladder Inflammation

LPS-肽在膀胱炎症中的相互作用

• 批准号: 6616412
• 负责人: RICARDO SABAN
• 金额: $ 32.23万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-29 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6616412
• 关键词: Escherichia coli; RNase protection assay; acute disease /disorder; chronic disease /disorder; endopeptidases; endotoxins; enzyme activity; enzyme inhibitors; enzyme reconstitution; gene targeting; genetically modified animals; immunocytochemistry; in situ hybridization; inflammation; interstitial cystitis; laboratory mouse; lipopolysaccharides; mast cell; molecular pathology; neuropeptide receptor; substance P; urinary bladder disorder; western blottings

DESCRIPTION (provided by applicant): This proposal is a competing continuation of a four year grant which seeks to expand upon the role of mast cells and mast cell products, substance P (SP), and LPS in bladder inflammation and further elucidate the molecular pathways resulting in up-regulation of inflammatory genes. The original specific aims were to determine the role of neutral metalloendopeptidase (NEP), SP receptors, and mast cells in the development of cystitis, utilizing NEP -/- mice, NK-1R -/- mice, and mast cell deficient mice (kit[w]/kit[w-v]), and those which have been reconstituted with mast cells by adoptive transfer from normal mice and NK-1R -/- mice. In the present proposal, we seek to expand upon the hypothesis that regardless of the stimuli there is a common intracellular activation pathway of bladder inflammation. We now include the possible role of Protease Activated-Receptors (PAR) as a signaling mechanism owing to tryptase release following mast cell degranulation. There are 6 specific aims in this proposal seeking to determine the role of PARs in the development of acute and chronic bladder inflammation. We will employ methodology and animal models published by this laboratory, including determination of cellular pattern of the 4 different PAR proteins and their gene expression by immunohistochemistry and in situ hybridization. Results will be confirmed by western blotting and quantifiable RNase protection assay developed by this laboratory. The role of PAR in inflammation will be confirmed in PAR -/- mice. In addition, the role of PARs 1 and 2 on mast cells will be determined by examining kit[w]/kit[w-v] mice, which are deficient in mast cells, after they receive bone-marrow mast cells from PAR1-/- or PAR2-/- mice. This study seeks to determine the molecular pathway of acute and chronic bladder inflammation. Information generated by this study should elucidate whether bladder inflammatory responses, regardless of etiology, occur via a common mechanism or whether the endpoints are specific for each stimulus.

描述（由申请人提供）：该提案是一项为期四年的资助的竞争性延续，该资助旨在扩大肥大细胞和肥大细胞产物、P物质（SP）和LPS在膀胱炎症中的作用，并进一步阐明导致炎症基因上调的分子途径。最初的具体目的是确定中性金属内肽酶（NEP）、SP受体和肥大细胞在膀胱炎发展中的作用，使用NEP -/-小鼠、NK-1 R-/-小鼠和肥大细胞缺陷小鼠（kit[w]/kit[w-v]），以及通过过继转移从正常小鼠和NK-1 R-/-小鼠用肥大细胞重建的小鼠。在本提案中，我们试图扩大的假设，无论刺激有一个共同的细胞内激活途径的膀胱炎症。我们现在包括蛋白酶激活受体（PAR）的可能作用作为一种信号机制，由于类胰蛋白酶释放肥大细胞脱粒。该提案中有6个具体目标，旨在确定PAR在急性和慢性膀胱炎症发展中的作用。我们将采用本实验室发表的方法和动物模型，包括通过免疫组织化学和原位杂交确定4种不同PAR蛋白的细胞模式及其基因表达。结果将通过本实验室开发的蛋白质印迹法和可定量的RNA酶保护试验进行确认。PAR在炎症中的作用将在PAR -/-小鼠中得到证实。此外，PAR 1和2对肥大细胞的作用将通过在接受来自PAR 1-/-或PAR 2-/-小鼠的骨髓肥大细胞后检查缺乏肥大细胞的kit[w]/kit[w-v]小鼠来确定。本研究旨在确定急性和慢性膀胱炎症的分子途径。本研究产生的信息应阐明膀胱炎症反应，无论病因如何，是否通过共同机制发生，或者终点是否对每种刺激具有特异性。