IL1 AND GINGIVAL COLLAGENASE GENE REGULATION

白细胞介素-1（IL-1）刺激牙龈成纤维细胞产生 金属蛋白酶（e . G.胶原酶、基质溶解素（蛋白聚糖酶））和 因此与牙龈软组织破坏有关， 牙周炎与IL-1牙龈炎相关的细胞内生化事件 成纤维细胞受体激活到金属蛋白酶基因激活是 不太了解。很可能是一些蛋白质（例如， 转录前/转录后调节因子、酶 参与信号转导）在这些调节中起关键作用。 金属蛋白酶滑膜和牙龈成纤维细胞的最新研究 在文化中提供了新的见解的分子事件相关的 随着IL-1对细胞金属蛋白酶的诱导， 这些金属蛋白酶的诱导信息， 牙龈成纤维细胞 在此基础上，本提案的目标 包括一系列的实验，旨在开始，以确定六个牙龈 IL-1诱导的成纤维细胞基因和产物可能参与了 可能与牙周炎相关的金属蛋白酶的调节。的 具体目的是：（1）获得四种IL-10基因的全长cDNA序列， 1诱导基因（以前未鉴定）从人牙龈获得 成纤维细胞，表征它们并表达它们的蛋白质，（2）确定 如果/如何蛋白产物来源于这三个IL-1诱导基因和两个 其它已知的被IL-1上调的基因（C/EBP和NF κ B）在功能上结合 启动子区域或调节金属蛋白酶转录，和（3） 为了确定这些选择的基因是否在功能上参与IL-1 金属蛋白酶的诱导。这些目标将通过以下方式实现： (a)牙龈成纤维细胞系和来自 从正常人和牙周炎患者获得的牙龈组织;（B） 全长cDNA克隆分离、表征和测序;（c） 计算机化序列分析;（d）体外蛋白质翻译;（e）共- 转染、过表达和低表达（反义）研究;以及 (f)凝胶移位结合和足迹分析。这些拟议的体外 对人牙龈成纤维细胞的研究提供了一种模型系统， 关于IL-1诱导的基因活化的基本分子信息，以及 从而鉴定可能参与发病机制的事件 牙周炎这些基因的鉴定有望改善 定义结缔组织破坏的分子基础， 牙周炎因此，一个重要的治疗意义， 目前的建议是，这些IL-1的阻断剂诱导的生化事件 为牙周炎的临床治疗提供了新的思路。

Interleukin-1 (IL-1) stimulates production of gingival fibroblast metalloproteinases (e . g. collagenase, stromelysin (proteoglycanase)) and thereby has been linked to gingival soft tissue destruction associated with periodontitis. The intracellular biochemical events that link IL-1 gingival fibroblast receptor activation to metalloproteinase gene activation are poorly understood. It is likely that a number of proteins (e.g. pretranscriptional/post-transcriptional regulatory factors, enzymes involved in signal transduction) play key roles in the modulation of these metalloproteinases. Recent studies with synovial and gingival fibroblasts in culture have provided new insights on the molecular events associated with the IL-1 induction of cellular metalloproteinases, there is much less information available on the induction of these metalloproteinases in gingival fibroblasts. Based on this, the objective of the present proposal involves a series of experiments designed to begin to identify six gingival fibroblast genes and products induced by IL-1 that could be involved in the modulation of metalloproteinases likely associated with periodontitis. The specific aims are to: (l) To obtain full length cDNA sequences for four IL- 1 induced genes (previously unidentified) obtained from human gingival fibroblasts, characterize them and express their proteins, (2) To determine if/how protein products derived from these three IL-1 induced genes and two other known genes (C/EBP and NFkB) upregulated by IL-1, functionally bind to the promoter region or modulate metalloproteinase transcription and (3) To determine if these selected genes functionally participate in the IL-1 induction of metalloproteinases. These aims will be accomplished by using: (a) gingival fibroblast cell lines and primary cultures derived from gingival tissue obtained from normal and patients with periodontitis; (b) full length cDNA clone isolation, characterization and sequencing; (c) computerized sequence analysis; (d) in vitro protein translation; (e) co- transfection, overexpression and underexpression (anti sense) studies; and (f) gel shift binding and foot printing assays. These proposed in-vitro studies with human gingival fibroblasts offer a model system to provide fundamental molecular information on gene activation induced by IL-1, and thereby identifying events which are likely involved in the pathogenesis of periodontitis. Identification of such genes holds promise for better defining the molecular basis of connective tissue destruction in periodontitis. Consequently, an important therapeutic implication of the present proposal is that blockers of these IL-1 induced biochemical events could be useful clinically for the future treatment of periodontitis.