激素干预对大鼠成骨细胞miRNA表达的影响

发布时间：2018-09-16 20:11

【摘要】：目的探讨正常与激素干预的大鼠成骨细胞的差异表达miRNA,并预测其靶基因。方法取SD乳鼠的成骨细胞进行培养、传代,取第3代成骨细胞分为空白组和激素组,空白组用含10%大鼠血清的L-DMEM培养基培养,激素组用含10~(-5)mol/L甲泼尼龙、10%大鼠血清的L-DMEM培养基;分别提取总RNA并进行质检;采用RNA标记与芯片杂交,采集数据及标准化,找出两组间的差异表达miRNA并对其表达谱进行分析。利用miranda、microcosm、mirdb 3个靶点预测软件对最明显的miRNA进行靶基因预测。结果与空白组相比,激素组中倍数2.0(表达上调)的miRNA有4个,倍数0.5(表达下调)的miRNA有2个,其中以上调的miR-672-5p和下调的miR-146a-5p最明显,其分别具有4个和11个可能靶基因。结论激素干预后大鼠成骨细胞的miRNA表达发生了明显变化,以miR-672-5p和miR-146a-5p最为显著。这表明它们有可能参与成骨细胞的凋亡和股骨头坏死的发生。
[Abstract]:Objective to investigate the differential expression of miRNA, and predict its target gene in osteoblasts of normal and steroid treated rats. Methods the osteoblasts of SD rats were cultured and subcultured. The third generation of osteoblasts were divided into blank group and hormone group. The blank group was cultured on L-DMEM medium containing 10% rat serum, and the hormone group was cultured on L-DMEM medium containing 10 ~ (-5) mol/L methylprednisolone 10% rat serum. The total RNA was extracted and quality checked, and the RNA marker and chip hybridization were used to collect the data and standardize the difference between the two groups to find out the difference of miRNA expression and analyze its expression profile. The target genes of the most obvious miRNA were predicted by using three target prediction software of miranda,microcosm,mirdb. Results compared with the control group, there were 4 miRNA (up-regulated) and 2 miRNA (down-regulated) miRNA in the hormone group. The up-regulated miR-672-5p and down-regulated miR-146a-5p were the most obvious, with 4 and 11 possible target genes, respectively. Conclusion the expression of miRNA in osteoblasts was significantly changed after hormone intervention, especially miR-672-5p and miR-146a-5p. This suggests that they may be involved in osteoblast apoptosis and osteonecrosis of the femoral head.
【作者单位】：广州中医药大学第一临床医学院;广州中医药大学第一附属医院骨科;
【基金】：国家自然科学基金资助项目(编号:81373652)
【分类号】：R681.8

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