miR-31 通过促进骨髓间充质干细胞的迁移来改善小鼠的脊髓损伤。
MiR-31 improves spinal cord injury in mice by promoting the migration of bone marrow mesenchymal stem cells.
机构信息
Department of Laboratory Animal Center, Shanxi Medical University, Taiyuan, Shanxi, China.
Department of Key Laboratory of Oral Disease Prevention and New Materials, Taiyuan, Shanxi, China.
出版信息
PLoS One. 2022 Sep 6;17(9):e0272499. doi: 10.1371/journal.pone.0272499. eCollection 2022.
BACKGROUND
Stem cell transplantation therapy is a potential approach for the repair of spinal cord injuries and other neurodegenerative diseases, but its effectiveness is hampered by the low rate of targeted migration of cells to the area of injury. The aim of this study was to investigate the effects of miR-31 on the migration of bone marrow mesenchymal stem cells (BMSCs) and the regulation of MMP-2 and CXCR4 expression in vitro and in vivo.
METHODS
eGFP-expressing BMSCs were isolated and cultured for subsequent experiments. The experiments were divided into three groups: control group, miR-31agomir group, and miR-31antagomir group. Proliferation was analyzed using CCK-8 and flow cytometry; cell migration in vitro was analyzed using wound-healing and transwell assays. The mouse SCI model was prepared by the impact method, and cells were transplanted (3 groups, 12 per group). Relevant inflammatory factors were detected by ELISA. The BMS score was used to evaluate the functional recovery of the mouse spinal cord and the frozen section was used to analyze the cell migration ability in vivo. The in vitro and in vivo expression levels of MMP-2 and CXCR4 were evaluated by Western blot and immunohistochemical staining.
RESULTS
In vitro experiments showed that cells in the miR-31agomir group exhibited enhanced cell proliferation (P<0.05, P<0.001) and migration (P<0.001) and upregulated protein expression levels of CXCR4 (P<0.01) and MMP-2 (P<0.001) compared with cells in the control group. The results of in vivo experiments showed that the expression of pro-inflammatory factors was reduced after cell transplantation treatment. Cells in the miR-31agomir group showed enhanced cell-targeted migration ability (P<0.001), improved the function of damaged tissues (P<0.001), and upregulated CXCR4 and MMP-2 expression compared to the control group (P<0.001).
CONCLUSION
Our experiment demonstrated that miR-31 could promote the migration of BMSCs and miR-31 could repair and improve the function of damaged tissues in SCI.
背景
干细胞移植疗法是修复脊髓损伤和其他神经退行性疾病的一种有潜力的方法，但由于细胞靶向迁移到损伤区域的效率低，其效果受到了限制。本研究旨在探讨 miR-31 对骨髓间充质干细胞（BMSCs）迁移的影响及其对 MMP-2 和 CXCR4 表达的体内外调节作用。
方法
分离和培养 eGFP 表达的 BMSCs 进行后续实验。实验分为三组：对照组、miR-31agomir 组和 miR-31antagomir 组。通过 CCK-8 和流式细胞术分析增殖；通过划痕愈合和 Transwell 分析体外细胞迁移。通过冲击法制备小鼠 SCI 模型，移植细胞（每组 12 只）。通过 ELISA 检测相关炎症因子。用 BMS 评分评估小鼠脊髓功能恢复情况，用冰冻切片分析体内细胞迁移能力。通过 Western blot 和免疫组化染色评估 MMP-2 和 CXCR4 的体内外表达水平。
结果
体外实验结果显示，miR-31agomir 组细胞增殖（P<0.05，P<0.001）和迁移（P<0.001）能力增强，CXCR4（P<0.01）和 MMP-2（P<0.001）蛋白表达水平升高。体内实验结果表明，细胞移植治疗后促炎因子表达减少。miR-31agomir 组细胞具有增强的靶向迁移能力（P<0.001），改善了受损组织的功能（P<0.001），并上调了 CXCR4 和 MMP-2 的表达（P<0.001）。
结论
本实验表明，miR-31 可促进 BMSCs 的迁移，miR-31 可修复和改善 SCI 中受损组织的功能。
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