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Altered microRNA profile during fracture healing in rats with diabetes
Shunsuke Takahara1,2, Sang Yang Lee1,3, Takashi Iwakura1
1Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, 650-0017, Japan.
Background:
MicroRNAs (miRNAs) are a class of small non-coding RNA molecules that regulate gene expression. There is increasing evidence that some miRNAs are involved in the pathology of diabetes mellitus (DM) and its complications. We hypothesized that the functions of certain miRNAs and the changes in their patterns of expression may contribute to the pathogenesis of impaired fractures due to DM.
Methods:
In this study, 108 male Sprague-Dawley rats were divided into DM and control groups. DM rats were created by a single intravenous injection of streptozotocin. Closed transverse femoral shaft fractures were created in both groups. On post-fracture days 5, 7, 11, 14, 21, and 28, miRNA was extracted from the newly generated tissue at the fracture site. Microarray analysis was conducted with miRNA samples from each group on post-fracture days 5 and 11. The microarray findings were validated by real-time polymerase chain reaction (PCR) analysis at each time point.
Results:
Microarray analysis revealed that, on days 5 and 11, 368 and 207 miRNAs, respectively, were upregulated in the DM group, compared with the control group. The top four miRNAs on day 5 were miR-339-3p, miR451-5p, miR-532-5p, and miR-551b-3p. The top four miRNAs on day 11 were miR-221-3p, miR376a-3p, miR-379-3p, and miR-379-5p. Among these miRNAs, miR-221-3p, miR-339-3p, miR-376a-3p, miR-379-5p, and miR-451-5p were validated by real-time PCR analysis. Furthermore, PCR analysis revealed that these five miRNAs were differentially expressed with dynamic expression patterns during fracture healing in the DM group, compared with the control group.
Conclusions:
Our findings will aid in understanding the pathology of impaired fracture healing in DM and may support the development of molecular therapies using miRNAs for the treatment of impaired fracture healing in patients with DM.
Insights
This study identifies specific microRNAs (miRNAs) that are differentially expressed in diabetic rats with fractures, revealing key molecular players in impaired bone healing. These findings may lead to new miRNA-based therapies for diabetic fracture complications.
Area of Science:
- Molecular Biology
- Genetics
- Endocrinology
Background:
- MicroRNAs (miRNAs) are small non-coding RNAs regulating gene expression.
- Emerging evidence links specific miRNAs to diabetes mellitus (DM) pathology and complications.
- This study investigates the role of miRNA expression changes in impaired fracture healing in DM.
Purpose of the Study:
- To identify specific microRNAs (miRNAs) and their expression patterns in diabetic rats with femoral fractures.
- To understand the contribution of altered miRNA function to the pathogenesis of impaired fracture healing in diabetes mellitus.
Main Methods:
- 108 male Sprague-Dawley rats were divided into diabetic (DM) and control groups.
- Femoral shaft fractures were induced; miRNA was extracted from fracture sites at multiple time points.
- Microarray analysis identified differentially expressed miRNAs, validated by real-time PCR.
Main Results:
- Microarray analysis revealed significant upregulation of 368 miRNAs (day 5) and 207 miRNAs (day 11) in DM rats compared to controls.
- Key upregulated miRNAs identified include miR-339-3p, miR-451-5p, miR-532-5p, miR-551b-3p (day 5) and miR-221-3p, miR-376a-3p, miR-379-3p, miR-379-5p (day 11).
- Real-time PCR validated differential expression of miR-221-3p, miR-339-3p, miR-376a-3p, miR-379-5p, and miR-451-5p during fracture healing in DM rats.
Conclusions:
- The identified miRNAs exhibit dynamic expression patterns during fracture healing in diabetic rats.
- These findings enhance understanding of impaired fracture healing in DM.
- The results may inform the development of novel miRNA-based molecular therapies for diabetic fracture complications.
