本文關(guān)鍵詞： 成骨分化 轉(zhuǎn)錄因子 Runx2 Osterix HDAC TSA　出處：《東北師范大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 在當(dāng)今社會(huì),骨骼疾病越來(lái)越受到人們的關(guān)注,因此對(duì)于骨的研究也就越來(lái)越深入。骨是一個(gè)動(dòng)態(tài)的、復(fù)雜的組織,在它的形成以及發(fā)揮功能的過(guò)程中,受到了各種轉(zhuǎn)錄因子、信號(hào)通路、染色體改構(gòu)復(fù)合物等因素的影響。其中Runx2和Osterix兩種轉(zhuǎn)錄因子在成骨細(xì)胞分化、骨的發(fā)育、骨形成過(guò)程中起著決定性的作用,Osterix位于Runx2的下游,是Runx2的靶基因。本文主要對(duì)在間充質(zhì)干細(xì)胞向成骨細(xì)胞分化過(guò)程中Osterix的作用做了初步的分析和研究。 通過(guò)ALP染色和von Kossa染色及RT-PCR實(shí)驗(yàn),我們發(fā)現(xiàn)在過(guò)表達(dá)Osterix的細(xì)胞中,Osterix可以促進(jìn)間充質(zhì)干細(xì)胞向成骨細(xì)胞分化,但是它的促分化能力卻不及Runx2強(qiáng),即使加入TSA, Osterix對(duì)成骨細(xì)胞分化影響也沒(méi)有太大的變化。接著通過(guò)RT-PCR實(shí)驗(yàn),我們檢驗(yàn)到在間充質(zhì)干細(xì)胞向成骨細(xì)胞分化的第6天,過(guò)表達(dá)Runx2的細(xì)胞中某些成骨細(xì)胞分化過(guò)程中標(biāo)志性基因的表達(dá)在TSA的作用下發(fā)生了變化如骨涎蛋白(BSP)、骨鈣素(OCN)等;而在過(guò)表達(dá)Osterix的細(xì)胞系中標(biāo)志性基因表達(dá)水平的改變并不像過(guò)表達(dá)Runx2的細(xì)胞系那么顯著,有時(shí)候甚至?xí)霈F(xiàn)與過(guò)表達(dá)Runx2的細(xì)胞系相反的現(xiàn)象。所以HDAC對(duì)Osterix的作用也許并沒(méi)有我們所想象的那么明顯或是那么簡(jiǎn)單,組蛋白去乙�；�(HDAC)對(duì)成骨細(xì)胞的影響可能主要還是通過(guò)影響Runx2來(lái)實(shí)現(xiàn)的。 我們的結(jié)果表明Osterix對(duì)成骨細(xì)胞的分化有一定的促進(jìn)作用,但是作用要比Runx2弱。而在間充質(zhì)干細(xì)胞向成骨細(xì)胞分化過(guò)程中,我們初步探索得到的結(jié)果是HDAC對(duì)Osterix的作用并沒(méi)有我們所希望的那么顯著,還需要在未來(lái)的實(shí)驗(yàn)中不斷的探索。
[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.
【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329

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