MicroRNAs调控人脂肪源性干细胞成骨分化及ODN影响其调控的研究

发布时间：2018-03-01 08:11

  本文关键词： 脂肪源性干细胞 成骨诱导 多分化潜能 microRNA 寡核苷酸　出处：《吉林大学》2010年博士论文　论文类型：学位论文

【摘要】： 本实验的目的是识别miRNAs是人脂肪源性干细胞(human Adipose Derived Stem Cells,hADSCs)成骨分化的一个调控因子,探讨对成骨分化有调节作用的寡核苷酸(oligodeoxynucleotide,ODN)是否通过调控miRNAs的表达从而调节成骨分化。 从人脂肪抽吸物中分离hADSCs,体外培养扩增及鉴定。对hADSCs成骨分化的效率进行评估。检测miRNA-196a、microRNA-133a在hADSCs成骨分化中的表达以及Runx2、HOXC8的蛋白表达。将ODN C2及C12加入hADSCs成骨诱导培养基中诱导hADSCs成骨分化,以成骨诱导培养基诱导的hADSCs作为对照。检测三组成骨能力的变化,miRNA-196a、microRNA-133a表达的变化,Runx2、HOXC8蛋白表达的变化。 结果证明hADSCs有极强的向成骨细胞分化的能力。miRNA-196a的表达在hADSCs成骨分化的过程中增强,伴随着HOXC8蛋白表达降低。提示miRNA-196a在hADSCs成骨分化中有重要作用,它的作用是通过HOXC8这一靶点介导的。通过本实验,miRNA-133a对成骨分化是否有调控作用尚不十分明确。本实验结果显示ODN C2及C12对成骨有抑制作用,是通过下调miRNA-196a表达水平,增加HOXC8的蛋白表达来实现的。 本实验研究证明miRNAs是hADSCs成骨分化的调控因子;首次将ODN应用于hADSCs的成骨分化过程中,显示其对hADSCs的成骨分化有抑制作用;并证明ODN C2及C12对成骨的抑制作用是通过下调miRNA-196a表达水平,增加HOXC8的蛋白表达实现的。为hADSCs应用于临床作为组织工程骨的种子细胞及探索对成骨分化有影响的药物开发提供新的思路。
[Abstract]:The purpose of this study was to identify miRNAs as a regulatory factor for osteogenic differentiation of human Adipose / Derived Stem cells Stem ADSCs, and to investigate whether the oligodeoxynucleotideODNs can regulate osteogenic differentiation by regulating the expression of miRNAs. HADSCswere isolated from human adipose extracts, cultured and identified in vitro. The efficiency of hADSCs osteogenic differentiation was evaluated. The expression of miRNA-196amicroRNA-133a in hADSCs osteogenic differentiation and the expression of Runx2HOXC8 protein were detected. ODN C2 and C12 were added to hADSCs osteogenesis induction. HADSCs osteogenic differentiation was induced in culture medium. HADSCs induced by osteoblast induction medium was used as control. The changes of osteogenic ability of the three groups were detected. The expression of microRNA-133a was detected and the protein expression of Runx2HoxC8 was detected. The results showed that hADSCs had a strong ability to differentiate into osteoblasts. The expression of miRNA-196a was enhanced during the osteogenic differentiation of hADSCs, and the expression of HOXC8 protein was decreased, suggesting that miRNA-196a plays an important role in the osteogenic differentiation of hADSCs. Its effect is mediated by the target of HOXC8. It is not clear whether miRNA-133a can regulate the osteogenic differentiation. The results show that ODN C2 and C12 inhibit osteogenesis and down-regulate the expression of miRNA-196a. Increase HOXC8 protein expression to achieve. This study proved that miRNAs is the regulatory factor of hADSCs osteogenic differentiation, and that ODN was used in the process of hADSCs osteogenic differentiation for the first time, which showed that ODN could inhibit the osteogenic differentiation of hADSCs. It was proved that the inhibitory effect of ODN C2 and C12 on osteogenesis was mediated by down-regulation of miRNA-196a expression. It provides a new idea for the application of hADSCs in clinical seed cells of tissue engineering bone and the development of drugs that have an effect on osteogenic differentiation.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R329

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