论文部分内容阅读
目的:成骨细胞和骨细胞凋亡学说是激素性股骨头坏死的机制之一,袁氏生脉成骨片已被证实在临床治疗激素性股骨头坏死有一定疗效,但两者具体作用机制有待深入,实验研究袁氏生脉成骨片对激素干预后大鼠成骨细胞的差异表达miRNA,预测其靶基因以推测作用机制。方法:取乳鼠(大鼠)的成骨细胞进行培养、传代,将第三代成骨细胞分为空白组,激素组,中药组;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,找出三组间的差异表达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miR-672-5p进行靶基因预测。结果:和空白组相比,激素组中倍数>2.0倍的miRNA共有6个(4个miRNA表达上调,2个miRNA表达下调);和空白组相比,中药组中倍数>2.0倍miRNA共有63个(19个miRNA表达上调,44个miRNA表达下调);和激素组相比,中药组中倍数>2.0倍的miRNA共有56个(11个miRNA表达上调,45个miRNA表达下调)。在激素组与空白组对照中上调,在中药组与激素组对照中相对应下降的倍数>2.0倍的miRNA共4个(miR-672-5p、miR-3559-3p、miR-98-3p、miR-92a-1-5p),并预测出Angptl4、Ccdc51、RGD1306991、Ssbp3作为miR-672-5p的靶基因。结论:袁氏生脉成骨片对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最为显著。这表明了它有可能参与了成骨细胞的凋亡,为袁氏生脉成骨片治疗激素性股骨头坏死提供理论依据。
OBJECTIVE: Osteoblasts and osteoblast apoptosis theory is one of the mechanisms of steroid-induced osteonecrosis of the femoral head. Yuan’s Shengmai osteogenic bone tablets have been proved to be effective in the treatment of steroid-induced osteonecrosis of the femoral head. However, both of them have a specific mechanism of action To be in-depth, experimental study of Yuan’s Shengmai Changgou piece of hormone-induced rat osteoblast differentially expressed miRNA, prediction of its target gene to speculate mechanism. Methods: Osteoblasts from neonatal rats (rat) were cultured and passaged. The third generation of osteoblasts were divided into blank group, hormone group and traditional Chinese medicine group. Total RNA was extracted and quantified. Hybridization, data collection and standardization, to identify differences between the three groups expressed miRNA and its expression profile analysis. Three target prediction softwares, miranda, microcosm and mirdb, were used to predict the most obvious target gene of miR-672-5p. Results: Compared with the blank group, the number of miRNAs in the hormone group was> 2.0 times that in the blank group (6 miRNAs were up-regulated and 2 miRNAs were down-regulated). Compared with the blank group, (19 miRNAs were up-regulated and 44 miRNAs were down-regulated). Compared with the hormone group, there were 56 miRNAs (> 11 miRNAs were up-regulated and 45 miRNAs were down-regulated). In the hormone group and the control group were up-regulated in the traditional Chinese medicine group and hormone control group corresponding to a multiple of 2.0 times the miRNA total of 4 (miR-672-5p, miR-3559-3p, miR-98-3p, miR-92a-1-5p) and predict Angptl4, Ccdc51, RGD1306991, Ssbp3 as target genes of miR-672-5p. Conclusion: YuanShengShengmaiMengGao miRNA expression of rat osteoblasts after hormone intervention significantly changed, with miR-672-5p the most significant. This indicates that it is possible to participate in the apoptosis of osteoblasts and provide a theoretical basis for the treatment of steroid-induced osteonecrosis of the femoral head by Yuan’s Shengmai Oral Bone.