CKIP-1对氟致成骨细胞凋亡的调控作用研究

项目名称： CKIP-1对氟致成骨细胞凋亡的调控作用研究

项目编号： No.31272623

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 韩博

作者单位： 中国农业大学

项目金额： 77万元

中文摘要： 近年来，本研究团队通过分子生物学和细胞生物学方法进行了一系列的氟对成骨细胞（OB）细胞因子表达和OB凋亡的影响研究，具备良好的研究基础。而CKIP-1因子于近期被证实对成骨细胞凋亡具有正向调控作用。因此本研究旨在通过RNAi技术干扰成骨细胞CKIP-1基因的表达，进行CKIP-1对体外氟致成骨细胞增殖凋亡的调控作用研究。研究内容包括：①建立OB原代培养模型并进行鉴定；②实时荧光定量PCR和Western-blot对氟致OB凋亡过程中CKIP-1基因在 mRNA和蛋白水平的表达；③用MTT法、电镜技术研究OB凋亡过程中其增殖活力和形态学的变化；④流式细胞术分析OB增殖周期和凋亡的变化；⑤利用电子顺磁共振波谱仪测定OB凋亡过程中自由基的变化。通过以上研究，阐明CKIP-1在氟致成骨细胞增殖凋亡过程中发挥的作用，从而为CKIP-1经过干扰表达技术应用于临床防治氟骨症提供依据。

中文关键词： CKIP-1；成骨细胞；凋亡；氟；调控

英文摘要： The casein kinase 2 interacting protein-1 (CKIP-1) plays an important role in regulation of bone formation and muscle differentiation. However, whether and how CKIP-1 activity is regulated on osteoblast apoptosis induced by fluoride remains largely unclear. The objective of this program is to evaluate the regulation of CKIP-1 on osteoblast apoptosis induced by fluoride. Currently, our research group has studied the effect of fluorine on cytokine expression and apoptosis of osteoblasts for a long time, it was proved that herein we have obtained good groundwork for the osteoblasts related research. The CKIP-1 has recently been demonstrated to express positive regulatory action for apoptosis of osteoblasts. So, we combined these research findings together to clarify the role of CKIP-1 when it join in the apoptosis of osteoblast caused by fluorine using RNAi assay. The contents of our design hereby are as following: (1) cultivation and identification of osteoblasts of ovine in vitro; (2) determination of both mRNA and protein expression of CKIP-1 during the process of apoptosis caused by fluoride using real time-PCR and western-blot; (3) determining the proliferation vitality and observing the morphology characteristics of osteoblasts by MTT assay and electron microscope; (4) analyzing the generation cycle and apopt

英文关键词： CKIP-1；Osteoblast；Apoptosis；Fluoride；Regulation