Takahara S, Lee S Y, Iwakura T, Oe K, Fukui T, Okumachi E, Waki T, Arakura M, Sakai Y, Nishida K, Kuroda R, Niikura T
Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 650-0017 Kobe, Japan.
Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine and Showa University School of Medicine,650-0017 Kobe, Japan and 142-8666 Tokyo, Japan.
Bone Joint Res. 2018 Feb;7(2):139-147. doi: 10.1302/2046-3758.72.BJR-2017-0082.R1.
Diabetes mellitus (DM) is known to impair fracture healing. Increasing evidence suggests that some microRNA (miRNA) is involved in the pathophysiology of diabetes and its complications. We hypothesized that the functions of miRNA and changes to their patterns of expression may be implicated in the pathogenesis of impaired fracture healing in DM.
Closed transverse fractures were created in the femurs of 116 rats, with half assigned to the DM group and half assigned to the control group. Rats with DM were induced by a single intraperitoneal injection of streptozotocin. At post-fracture days five, seven, 11, 14, 21, and 28, miRNA was extracted from the newly generated tissue at the fracture site. Microarray analysis was performed with miRNA samples from each group on post-fracture days five and 11. For further analysis, real-time polymerase chain reaction (PCR) analysis was performed at each timepoint.
Microarray analysis showed that there were 14 miRNAs at day five and 17 miRNAs at day 11, with a greater than twofold change in the DM group compared with the control group. Among these types of miRNA, five were selected based on a comparative and extended literature review. Real-time PCR analysis revealed that five types of miRNA (miR-140-3p, miR-140-5p, miR-181a-1-3p, miR-210-3p, and miR-222-3p) were differentially expressed with changing patterns of expression during fracture healing in diabetic rats compared with controls.
Our findings provide information to further understand the pathology of impaired fracture healing in a diabetic rat model. These results may allow the potential development of molecular therapy using miRNA for the treatment of impaired fracture healing in patients with DM.: S. Takahara, S. Y. Lee, T. Iwakura, K. Oe, T. Fukui, E. Okumachi, T. Waki, M. Arakura, Y. Sakai, K. Nishida, R. Kuroda, T. Niikura. Altered expression of microRNA during fracture healing in diabetic rats. 2018;7:139-147. DOI: 10.1302/2046-3758.72.BJR-2017-0082.R1.
已知糖尿病（DM）会损害骨折愈合。越来越多的证据表明，某些微小RNA（miRNA）参与糖尿病及其并发症的病理生理过程。我们推测，miRNA的功能及其表达模式的变化可能与糖尿病患者骨折愈合受损的发病机制有关。
在116只大鼠的股骨上造成闭合性横断骨折，其中一半分配到糖尿病组，另一半分配到对照组。通过单次腹腔注射链脲佐菌素诱导糖尿病大鼠。在骨折后第5、7、11、14、21和28天，从骨折部位新生成的组织中提取miRNA。在骨折后第5天和第11天，对每组的miRNA样本进行微阵列分析。为了进一步分析，在每个时间点进行实时聚合酶链反应（PCR）分析。
微阵列分析显示，在第5天有14种miRNA，在第11天有17种miRNA，糖尿病组与对照组相比变化超过两倍。在这些类型的miRNA中，基于比较和扩展的文献综述选择了5种。实时PCR分析显示，与对照组相比，糖尿病大鼠骨折愈合过程中5种miRNA（miR-140-3p、miR-140-5p、miR-181a-1-3p、miR-210-3p和miR-222-3p）的表达随时间变化存在差异。
我们的研究结果为进一步了解糖尿病大鼠模型中骨折愈合受损的病理提供了信息。这些结果可能有助于开发利用miRNA的分子疗法来治疗糖尿病患者骨折愈合受损。：S. Takahara、S.Y. Lee、T. Iwakura、K. Oe、T. Fukui、E. Okumachi、T. Waki、M. Arakura、Y. Sakai K. Nishida、R. Kuroda、T. Niikura。糖尿病大鼠骨折愈合过程中微小RNA的表达改变。2018；7：139 - 147。DOI：10.1302/2046 - 3758.72.BJR - 2017 - 0082.R1。
