Osterix在间充质干细胞向成骨细胞分化过程中作用的初步研究

发布时间：2018-02-27 07:28

本文关键词： 成骨分化转录因子 Runx2 Osterix HDAC TSA　出处：《东北师范大学》2010年硕士论文　论文类型：学位论文

【摘要】： 在当今社会,骨骼疾病越来越受到人们的关注,因此对于骨的研究也就越来越深入。骨是一个动态的、复杂的组织,在它的形成以及发挥功能的过程中,受到了各种转录因子、信号通路、染色体改构复合物等因素的影响。其中Runx2和Osterix两种转录因子在成骨细胞分化、骨的发育、骨形成过程中起着决定性的作用,Osterix位于Runx2的下游,是Runx2的靶基因。本文主要对在间充质干细胞向成骨细胞分化过程中Osterix的作用做了初步的分析和研究。通过ALP染色和von Kossa染色及RT-PCR实验,我们发现在过表达Osterix的细胞中,Osterix可以促进间充质干细胞向成骨细胞分化,但是它的促分化能力却不及Runx2强,即使加入TSA, Osterix对成骨细胞分化影响也没有太大的变化。接着通过RT-PCR实验,我们检验到在间充质干细胞向成骨细胞分化的第6天,过表达Runx2的细胞中某些成骨细胞分化过程中标志性基因的表达在TSA的作用下发生了变化如骨涎蛋白(BSP)、骨钙素(OCN)等;而在过表达Osterix的细胞系中标志性基因表达水平的改变并不像过表达Runx2的细胞系那么显著,有时候甚至会出现与过表达Runx2的细胞系相反的现象。所以HDAC对Osterix的作用也许并没有我们所想象的那么明显或是那么简单,组蛋白去乙酰化酶(HDAC)对成骨细胞的影响可能主要还是通过影响Runx2来实现的。我们的结果表明Osterix对成骨细胞的分化有一定的促进作用,但是作用要比Runx2弱。而在间充质干细胞向成骨细胞分化过程中,我们初步探索得到的结果是HDAC对Osterix的作用并没有我们所希望的那么显著,还需要在未来的实验中不断的探索。
[Abstract]:In today's society, bone disease is getting more and more attention, so the study of bone is more and more in-depth. Bone is a dynamic and complex tissue, and in its formation and function, it has been subjected to various transcription factors. Among these factors, Runx2 and Osterix play a decisive role in osteoblast differentiation, bone development and bone formation. Osterix is located downstream of Runx2. It is a target gene of Runx2. The role of Osterix in the differentiation of mesenchymal stem cells into osteoblasts was studied in this paper. By means of ALP staining, von Kossa staining and RT-PCR assay, we found that Osterix can promote the differentiation of mesenchymal stem cells into osteoblasts, but its ability to promote differentiation is not as strong as Runx2. Even if TSA was added, the effect of Osterix on osteoblast differentiation was not much changed. Then, through RT-PCR experiment, we examined the differentiation of mesenchymal stem cells into osteoblasts on the 6th day. During the differentiation of some osteoblasts which overexpressed Runx2, the expression of some iconic genes changed under the action of TSA, such as bone sialoprotein BSPN, osteocalcin (OCN) and so on. The change in the expression level of the iconic gene in the over-expressed Osterix cell line was not as significant as that in the over-expressed Runx2 cell line. Sometimes even the opposite of the cell lines that express Runx2, so the effect of HDAC on Osterix may not be as obvious or as simple as we think. The effect of histone deacetylase (HDAC) on osteoblasts may be achieved mainly by affecting Runx2. Our results show that Osterix can promote the differentiation of osteoblasts to some extent, but the effect is weaker than that of Runx2. However, during the differentiation of mesenchymal stem cells into osteoblasts, Our initial findings are that the effect of HDAC on Osterix is not as significant as we would like, and will need to be explored in future experiments.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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