摘要小分子核糖核酸(microRNA)参与几乎所有的生理和病理过程,包括成骨细胞增殖和分化。骨是一个刚性和动态的器官,由破骨细胞和成骨细胞调节骨吸收和骨形成。这种平衡的中断会导致各种各样的疾病,例如骨质疏松症，骨质疏松症的骨丢失，会减少骨吸收和骨形成。因此,调节成骨细胞破骨细胞活动是一个治疗骨质疏松症主要的方法。小分子核糖核酸(microRNA)是一类单链在18到22 nt的核苷酸组成，对于分化、细胞命运、细胞凋亡和各疾病的发病机理，其非编码RNA在细胞有重要的作用。潜在的治疗骨疾病和生物功能的microRNA因此得到了更多的关注。然而，骨修复的缺陷并没有被很好的研究，所以本研究根据Bio-oss骨粉具有良好的生物相容性和成骨细胞系MC3T3E1可以定向分化为典型的成骨细胞的特点，对Bio-oss与成骨细胞系MC3T3E1共培养的相关基因分析，调查microRNA的作用和潜力，研究成骨细胞分化和血管生成调节机制。71958

Abstract Micrornas (mirna) involved in almost all of the physiological and pathological process of edge, including osteoblast proliferation and differentiation。Bone is a rigid and dynamic organs, by the osteoclasts and osteoblasts adjust bone resorption and bone formation。The interruption of this balance will lead to various diseases, such as osteoporosis, bone loss, osteoporosis will reduce bone resorption and bone formation。Therefore, regulating osteoblast osteoclast activity is a method for the treatment of osteoporosis is mainly。Micrornas (mirna) is a kind of single in 18 to 22 nt nucleotide composition, for differentiation, cell fate, cell apoptosis, and the pathogenesis of the disease, the non-coding RNA plays a key role in cells。Potential in the treatment of bone diseases and biological functions of mirnas thus attracted more attention。Bone repair defects, however, has not been very good, so this research according to the Bio - oss has good biological compatibility and bone ossification cell line can MC3T3E1 directional differentiation to the characteristics of the typical osteoblasts, the Bio - oss and ossification cell line MC3T3E1 co-culture related gene analysis, investigating the role of mirnas and potential, and research on osteoblast differentiation and angiogenesis regulation mechanism。

毕业论文关键词：小分子核糖核酸；成骨细胞；MC3T3E1；小鼠颅顶骨成骨细胞

Key words: microRNA; osteoblast; MC3T3E1; Bio-oss

目   录

引言 4

1。材料与方法 6

1。1实验材料 6

1。1。1主要试剂与仪器 6

1。2实验方法 6

1。2。1 成骨细胞系MC3T3E1复苏 6

1。2。2诱导分化 7

1。2。3 Bio-oss与MC3T3E1共培养 7

1。2。4 ALP染色 7

1。2。5 microRNA提取及检测 8

2。结果与分析 11

2。1实验结果 11

2。1 。1 RNA琼脂糖凝胶电泳 11

2。1。2 Nanodrop检测RNA的纯度测量操作 12

2。1。3 反转录合成cDNA实验操作步骤 12

2。2实验分析 13

3。讨论 14

参考文献 15

致谢 16