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.

白介素1（IL-1）刺激金属蛋白酶的产生（MMPS。 例如胶原酶和Stromelysin）通过牙龈成纤维细胞。并有 因此与与软组织破坏有关 牙周炎。虽然IL-1诱导MMP的精确机制 基因转录尚未完全阐明，有证据表明 IL-1的作用是由多种转录因子介导的 可以以复杂和细胞特定方式与启动子相互作用。 这 该提案的具体目的是：1）确定转录因子 特别与胶原酶的启动子区域和 IL-1刺激牙龈成纤维细胞后，斯特罗梅辛基因； 2） 确定在特定目的1中确定的因素的作用 调节在 对IL-1的响应； 3）进一步表征和克隆或净化任何 以前未知因素在特定的目标＃1中确定，并显示为 在调节MMP基因中起着重要作用。这些具体 目标将通过使用标准分子生物学来实现 包括电泳迁移率分析（EMSA），DNase在内的技术 足迹和瞬态转染实验系统地进行 剖析启动子以寻找调节器元素和结合 复合物。除了提供有关如何的重要新信息 MMP的生产受到调节。这些研究也可能确定新的 牙周炎治疗剂的潜在靶标。