本文關(guān)鍵詞： 脂肪源性干細胞 成骨誘導(dǎo) 多分化潛能 microRNA 寡核苷酸　出處：《吉林大學(xué)》2010年博士論文　論文類型：學(xué)位論文

【摘要】： 本實驗的目的是識別miRNAs是人脂肪源性干細胞(human Adipose Derived Stem Cells,hADSCs)成骨分化的一個調(diào)控因子,探討對成骨分化有調(diào)節(jié)作用的寡核苷酸(oligodeoxynucleotide,ODN)是否通過調(diào)控miRNAs的表達從而調(diào)節(jié)成骨分化。 從人脂肪抽吸物中分離hADSCs,體外培養(yǎng)擴增及鑒定。對hADSCs成骨分化的效率進行評估。檢測miRNA-196a、microRNA-133a在hADSCs成骨分化中的表達以及Runx2、HOXC8的蛋白表達。將ODN C2及C12加入hADSCs成骨誘導(dǎo)培養(yǎng)基中誘導(dǎo)hADSCs成骨分化,以成骨誘導(dǎo)培養(yǎng)基誘導(dǎo)的hADSCs作為對照。檢測三組成骨能力的變化,miRNA-196a、microRNA-133a表達的變化,Runx2、HOXC8蛋白表達的變化。 結(jié)果證明hADSCs有極強的向成骨細胞分化的能力。miRNA-196a的表達在hADSCs成骨分化的過程中增強,伴隨著HOXC8蛋白表達降低。提示miRNA-196a在hADSCs成骨分化中有重要作用,它的作用是通過HOXC8這一靶點介導(dǎo)的。通過本實驗,miRNA-133a對成骨分化是否有調(diào)控作用尚不十分明確。本實驗結(jié)果顯示ODN C2及C12對成骨有抑制作用,是通過下調(diào)miRNA-196a表達水平,增加HOXC8的蛋白表達來實現(xiàn)的。 本實驗研究證明miRNAs是hADSCs成骨分化的調(diào)控因子;首次將ODN應(yīng)用于hADSCs的成骨分化過程中,顯示其對hADSCs的成骨分化有抑制作用;并證明ODN C2及C12對成骨的抑制作用是通過下調(diào)miRNA-196a表達水平,增加HOXC8的蛋白表達實現(xiàn)的。為hADSCs應(yīng)用于臨床作為組織工程骨的種子細胞及探索對成骨分化有影響的藥物開發(fā)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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2010
【分類號】：R329

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