IL-1 REGULATION OF MMP GENES--HUMAN GINGIVAL FIBROBLASTS

IL-1对MMP基因的调控--人牙龈成纤维细胞

• 批准号: 2132941
• 负责人: RUTH C BORGHAEI
• 金额: $ 3.49万
• 依托单位: PHILADELPHIA COLLEGE OF OSTEOPATHIC MED
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-15 至 1997-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2132941
• 关键词: DNA binding protein; DNA footprinting; affinity chromatography; chloramphenicol acetyltransferase; collagenase; complementary DNA; crosslink; enzyme induction /repression; fibroblasts; gel mobility shift assay; genetic library; genetic promoter element; genetic regulation; gingiva; human subject; interleukin 1; ion exchange chromatography; molecular cloning; molecular site; protein purification; site directed mutagenesis; stromelysin; tissue /cell culture; transcription factor; transfection

Interleukin-1 (IL-1) stimulates production of metalloproteinases (MMPs. e.g. collagenase and stromelysin) by gingival fibroblasts. and has therefore been linked to soft tissue destruction associated with periodontitis. Although the precise mechanisms of IL-1 induction of MMP gene transcription have not been fully elucidated, evidence suggests that the effects of IL-1 are mediated by multiple transcription factors which may interact with the promoters in complex and cell specific ways. The specific aims of this proposal are: 1) to identify transcription factors that specifically interact with the promoter regions of collagenase and stromelysin genes following IL-1 stimulation of gingival fibroblasts; 2) to determine the role(s) of factors identified in Specific Aim #1 in regulating transcription of collagenase and/or stromelysin genes in response to IL-1; and 3) to further characterize and clone or purify any previously unknown factor(s) identified in Specific Aim #1 and shown to play a significant role in regulation of the MMP genes. These specific aims will be accomplished through the use of standard molecular biology techniques including electrophoretic mobility shift assays (EMSA), DNAse footprinting and transient transfection experiments to systematically dissect the promoters In search of regulator elements and binding complexes. In addition to providing important new information about how production of MMPs is regulated. these studies might also identify new potential targets for therapeutic agents for periodontitis.

白细胞介素-1（IL-1）刺激金属蛋白酶（MMPs）的产生。 例如胶原酶和基质溶解素）。并且具有 因此与软组织破坏有关， 牙周炎虽然IL-1诱导MMP的确切机制 基因转录尚未完全阐明，证据表明， IL-1的作用由多种转录因子介导， 可以以复杂的和细胞特异性的方式与启动子相互作用。 的 该建议的具体目标是：1）鉴定转录因子 与胶原酶的启动子区域特异性相互作用， IL-1刺激牙龈成纤维细胞后的基质溶解素基因; 2） 确定具体目标#1中确定的因素的作用 调节胶原酶和/或溶基质素基因在 对IL-1的应答;以及3）进一步表征和克隆或纯化任何 在具体目标#1中确定的先前未知因素， 在MMP基因的调控中起重要作用。这些具体 目标将通过使用标准分子生物学来实现 技术包括电泳迁移率变动分析（EMSA）、DNA酶 足迹法和瞬时转染实验系统地 剖析启动子寻找调节元件和结合 配合物除了提供关于如何 MMPs的产生受到调控。这些研究也可能发现新的 牙周炎治疗剂的潜在靶点。