CYTOKINE GENE REGULATION IN BONE REMODELING

骨重塑中的细胞因子基因调控

• 批准号: 2128816
• 负责人: KATHERINE A KELLY ABBOTT
• 金额: $ 6.5万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-30 至 1999-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2128816
• 关键词: bone development; bone marrow; cytokine; dimethylsulfoxide; gene expression; genetic regulation; hematopoietic stem cells; interleukin 6; interleukin 8; laboratory mouse; lipopolysaccharides; macrophage; messenger RNA; mixed tissue /cell culture; osteoblasts; osteoclasts; physiologic bone resorption; scanning electron microscopy; stainings; stromal cells; transcription factor; tumor necrosis factor alpha

This research project will address the process of bone remodelling, specifically the role of cytokines in the development and functioning of osteoclasts. In Aim 1, the cytokine gene regulation of activated macrophages will be studied in culture systems stimulated with lipopolysaccharide (LPS). The representative osteolytic cytokines tumor necrosis factor (TNF), macrophage inflammatory protein (MIP), and interleukin 6 (IL-6) will be studied to measure gene expression and activation of the transcriptional regulatory protein NF kappa B. The bioactivity of TNF and IL-6 produced by stimulated macrophages will be calculated. The data from the above studies will provide a foundation for the long- term bone marrow culture experiments in Aim 2. These studies will examine the communication network between hematopoietic/osteoclast progenitors and their supporting stromal/osteoblast cells during osteoclast maturation and function. Aim 2(a) will establish the optimal culture conditions for osteoclast-like cell enrichment, as well as methods to assay osteoclast- like cell number and function using tartrate resistant acid phosphatase staining and scanning electron microscopy. Aim 2(b) addresses the effect of LPS on osteoclast development in primary bone marrow cultures utilizing the LPS non-responsive murine strain C3H/Hej and the (LPS responsive) control C3H/Fej strain. Aim 2(c) examines the role of factors produced by LPS induced stromal/osteoblast cells on osteoclast development and function utilizing the C3H/Fej and C3H/Hej murine strains. These studies will take advantage of the well characterized stromal cell line derived from control (BMS2) and osteopetrotic (OP) mice.

该研究项目将解决骨骼重塑的过程， 特别是细胞因子在开发和功能中的作用 破骨细胞。 在AIM 1中，激活的细胞因子基因调节 将在刺激的培养系统中研究巨噬细胞 脂多糖（LPS）。 代表性骨化细胞因子肿瘤 坏死因子（TNF），巨噬细胞炎症蛋白（MIP）和 将研究白介素6（IL-6），以测量基因表达和 转录调节蛋白NF Kappa B的激活B. 受刺激巨噬细胞产生的TNF和IL-6的生物活性将是 计算。 上述研究的数据将为长期提供基础 AIM 2中的术语骨髓培养实验。这些研究将检查 造血/破骨细胞祖细胞与 它们在破骨细胞成熟期间的支撑基质/成骨细胞和 功能。 AIM 2（a）将建立最佳文化条件 破骨细胞样细胞富集以及测定破骨细胞的方法 例如使用抗tartrate抗酸性磷酸酶的细胞数和功能 染色和扫描电子显微镜。 目标2（b）解决效果 利用原代骨髓培养的破骨细胞发育的LPS的利用 LPS无反应性鼠菌株C3H/HEJ和（LPS响应） 控制C3H/FEJ菌株。 AIM 2（c）检查了由 LPS在破骨细胞发育和 利用C3H/FEJ和C3H/HEJ鼠菌株的功能。 这些研究 将利用衍生得出的良好特征的基质细胞系 从对照（BMS2）和骨质术（OP）小鼠中。