BONE CELL CALCIUM REGULATION

本研究的长期目标是了解 对骨细胞钙离子的调节。 内源性和外源性 将评估控制该参数的因素， 这些因素和钙之间的潜在相互关系， 评价骨代谢的调节。 的影响 甲状旁腺激素（PTH），前列腺素E2（PGE 2），表皮 生长因子（EGF）和白细胞介素-I（IL-1）将在 正常大鼠成骨细胞和骨肉瘤细胞系UMR-106 SaOS-2 的假定影响之间的相互关系 这些对钙和磷酸肌醇代谢的作用剂也将 通过研究这种磷脂的调节， 通过研究磷酸肌醇的作用， 在这些孤立的细胞系统中钙的动员。 花生四烯酸和胞浆钙之间的相互关系将 也被评估和花生四烯酸的影响， 还将研究细胞内钙动员。 的作用 蛋白激酶C在成骨细胞钙调节中的作用 代谢也将研究使用抑制剂和激活剂， 这种酶。 蛋白质、DNA合成和纤溶酶原的研究 还将进行活化剂试验， 细胞质钙离子的变化。 花生四烯酸代谢 也将在各种细胞系统中进行研究， 了解环氧合酶产物之间的相互关系 和胞质钙。 将测量细胞溶质钙水平 使用Fura 2荧光测定，细胞悬浮， 附在盖玻片上。 将研究磷酸肌醇代谢 采用薄层和离子交换色谱法， 高效液相色谱法 花生四烯酸代谢 还将通过高效液相色谱法进行研究。 这些研究应该能使我们更好地了解 骨细胞代谢和援助的治疗发展， 全身和局部骨代谢紊乱。

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.