BONE CELL CALCIUM REGULATION

骨细胞钙调节

• 批准号: 3155289
• 负责人: ROSEMARY Mary DZIAK
• 金额: $ 12.14万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3155289
• 关键词: arachidonate; bone metabolism disorder; calcium metabolism; cytoplasm; epidermal growth factor; fluorescence; fluorouracil; high performance liquid chromatography; homeostasis; hormone regulation /control mechanism; infant animal; interleukin 1; laboratory rat; osteoblasts; parathyroid hormones; phosphatidylinositols; phospholipids; plasminogen activator; prostaglandin E; prostaglandin endoperoxide synthase; protein kinase C; sarcoma; thin layer chromatography; tissue /cell culture

The long term objective of this study is to understand the nature of bone cell calcium regulation. The endogenous and exogenous factors that control this parameter will be assessed and the potential interrelationships between these factors and calcium in the regulation of bone metabolism will be evaluated. The effects of parathyroid hormone (PTH), prostaglandin E2 (PGE2), epidermal growth factor (EGF) and interleukin -I(IL-l) will be studied in normal rat osteoblastic cells and the osteosarcoma lines - UMR-106 and SaOS-2. The interrelationships between putative effects of these agents on calcium and phosphoinositide metabolism will also be studied by investigating the regulation of this phospholipid in the bone cells and by studying the effects of inositol phosphates on calcium mobilization in these isolated cell systems. Interrelationships between arachidonic and cytosolic calcium will also be evaluated and the effects of arachidonic acid on intracellular calcium mobilization will also be studied. The role of protein kinase C in the regulation of osteoblastic calcium metabolism will also be studied using inhibitors and activators of this enzyme. Studies on protein, DNA synthesis and plasminogen activator will also be conducted in an attempt to make correlations with changes in cytosolic calcium. Arachidonic acid metabolism will also be studied in the various cell systems to better understand the interrelationship between cyclooxygenase products and cytosolic calcium. Cytosolic calcium levels will be measured using the Fura 2 fluorescent assay with cells in suspension and attached to coverslips. Phosphoinositol metabolism will be studied with thin layer and ion exchange chromatography and high performance liquid chromatography. Arachidonic acid metabolism will also be studied by high performance liquid chromatography. These studies should lead to a better understanding of the nature of bone cell metabolism and aid in the development of therapy for systemic and localized bone metabolic disorders.

这项研究的长期目标是了解本质 骨细胞钙调节。 内源性和外源性 将评估控制该参数的因素，并 这些因素与钙之间的潜在相互关系 将评估骨代谢的调节。 效果 甲状旁腺激素（PTH）、前列腺素E2（PGE2）、表皮 生长因子（EGF）和白细胞介素-I（IL-l）将在 正常大鼠成骨细胞和骨肉瘤细胞系 - UMR-106 和SaOS-2。 假定的影响之间的相互关系 这些药物对钙和磷酸肌醇代谢的影响也会 通过研究这种磷脂的调节来研究 骨细胞并通过研究磷酸肌醇的作用 关于这些分离的细胞系统中的钙动员。 花生四烯酸和细胞质钙之间的相互关系 还评估了花生四烯酸对 细胞内钙动员也将被研究。 角色 蛋白激酶C在成骨细胞钙调节中的作用 还将使用抑制剂和激活剂来研究代谢 这种酶。 蛋白质、DNA合成和纤溶酶原的研究 还将进行激活器以尝试建立关联 随着细胞质钙的变化。 花生四烯酸代谢 还将在各种细胞系统中进行研究，以更好地 了解环氧合酶产物之间的相互关系 和细胞质钙。 将测量细胞质钙水平 使用 Fura 2 荧光测定法对悬浮细胞进行检测 附着在盖玻片上。 将研究磷酸肌醇代谢 具有薄层和离子交换色谱法以及高 高效液相色谱法。 花生四烯酸代谢 还将通过高效液相色谱进行研究。 这些研究应该有助于人们更好地了解自然 骨细胞代谢并有助于开发治疗方法 全身和局部骨代谢紊乱。