摘要
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： Apoptosis is one of the mechanisms of steroid induced ostenonecrosis of the femoral head and Yuan＇ s Shengmai Chenggu Tablet is a good treatment of steroid induced osteonecrosis of the femoral head in clinic. In this experiment ,we studied the differential expression of miRNA between osteoblasts after hormone intervention and that treated by Yuan＇ s Shengmai Chenggu Tablet to predict the target genes and possible mechanism of the Chinese medicine. Methods： The osteoblasts were cultured in rats and the third generation osteoblasts were divided into three groups：blank control group, hormone group, Chinese medicine group. The total RNA was extracted and the quality control was carried out. The data was collected by RNA marker and chip hybridization. Miranda, microcosm and mirdb target point prediction software were used for the most obvious miR - 672 - 5p target gene prediction. Results ： In comparison with the blank group, there was 6 miRNA （ 4 miRNA up - regulation and 2 miRNA down - regulation in the hormone group （ ratio 〉 2. 0 ）. In comparison with the blank group, there was 63 miRNA （ 19 miRNA up - regulation and 44 miRNA down - regu- lation in Chinese medicine group （ratio 〉 2. 0）. In comparison with the hormone group, there was 56 miRNA （11 miRNA up - regulation and 45 miRNA down - regulation in Chinese medicine group （ ratio 〉 2.0 ）. Up - regulation in the hor- mone group compared with blank control group while in the Chinese medicine group compared with hormone group, there was 4 miRNA （ miR - 672 - 5p, miR - 98 - 3p, miR - 3559 - 3p, miR - 92a - 1 - 5p）. Angptl4, Cede51, Ssbp3 and RGD1306991 were as the target gene of miR -672 -5p. Conclusion： The expression of miRNA in osteoblasts of rats after hoi^none intervention was significantly changed by Yuan＇ s Shengmai Chenggu Tablet and miR- 672 -5p was the most significant. This suggests that it may be involved in the apoptosis of osteoblasts ,which provides a theoretical basis for the treatment of steroid induced osteonecrosis of the femoral head by Yuan＇ s Shengmai Chenggu Tablet.
出处
《中华中医药学刊》
CAS
北大核心
2017年第1期91-94,I0004,共5页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金项目(81373652)
关键词
袁氏生脉成骨片
成骨细胞
激素性股骨头坏死
miR-672—5p
Yuan＇ s Shengmai Chenggu Tablet
osteoblasts
steroid induced osteonecrosis of the femoral head
miR - 672 - 5p
