我们发现成骨分化异常的BMSCs上Integrinα2、Notch1表达下调，且有密切的相互作用关系。因此拟进一步探讨Notch1调控Integrinα2影响成骨分化的分子机制及同OP的相关性，首先在人BMSCs成骨分化模型上采用了免疫共沉淀和激光共聚焦两种共定位方法分析它们的定位及关系，分化过程中的变化特点；进而在细胞及体内试验中分别上调或下调Notch1对Integrin α2活性的影响、信号通道相关分子的改变及对成骨分化能力的调节；我们引入邻位连接技术，检测Notch1调节Integrin α2下游复合体的机制；在骨质疏松患者BMSCs观察以上结构和通路在骨疏条件下的改变及意义；最后通过体内实验验证Notch1调控Integrin α2影响骨质疏松的机制。研究将阐明Notch1调节Integrin α2影响BMSCs成骨分化及骨质疏松的分子机制，并为药物研发提供新靶点。

We found that the decreased Integrin a2, Notch1 expression in BMSCs with abnormal osteogenic differentiation , and there is a close interaction. Accordingly, we propose to further investigate the role and mechanism of Notch1 regulating Integrin a2 on osteogenic differentiation of BMSCs and the relevance to OP.First,we will demonstrate the localization/structural changes by co-immunoprecipitation and laser scanning confocal in human BMSCs during osteogenesis differentiation model.Second,the changes of expression,signal passway,ability of regulate differentiation during Notch1 regulating Integrin a2 will be detected in vivo and vitro models;using the Proximity Ligation Assay system to identify proteins sites that interact with another protein;observing the changes of protein structure and passways in BMSCs that come from osteoporosis patients;verifying the molecular mechanisms of Notch1 regulating Integrin a2 on osteogenic differentiation of BMSCs and the relevance to OP in vivo models.Finally,the project will provide the significant theoretical basis for the research of osteogenic differentiation and OP,and offer new targets for the medicine research and development.