| 梁慧琦,李琳,杨琰,等.川芎嗪调控miR-139-5p/CXCR4通路促进骨髓间充质干细胞迁移[J].浙江中医药大学学报,2023,47(7):703-711. |
| 川芎嗪调控miR-139-5p/CXCR4通路促进骨髓间充质干细胞迁移 |
| Tetramethylpyrazine Promotes Bone Marrow Mesenchymal Stem Cells Migration by Regulating miR-139-5p/CXCR4 |
| DOI：10.16466/j.issn1005-5509.2023.07.001 |
| 中文关键词: 川芎嗪 骨髓间充质干细胞 细胞迁移 miR-139-5p CXCR4 脑缺血 |
| 英文关键词: tetramethylpyrazine BMSCs cell migration miR-139-5p CXCR4 cerebral ischemia |
| 基金项目:国家自然科学基金项目(81903949、81873028)；浙江省基础公益研究计划项目(LQ19H290004、2016C22185) |
|
| 摘要点击次数: 1508 |
| 全文下载次数: 915 |
| 中文摘要: |
| [目的] 探讨川芎嗪(tetramethylpyrazine，TMP)是否通过调控miR-139-5p/ 趋化因子受体4(chemokine receptor 4，CXCR4)促进骨髓间充质干细胞(bone marrow mesenchymal stem cells，BMSCs)迁移。[方法] 采用全骨髓贴壁法培养SD大鼠BMSCs，50、100 μmol·L-1的TMP预处理BMSCs 24 h，脂质体3000转染miR-139-5p mimic、inhibitor，以细胞划痕和Transwell实验检测BMSCs迁移能力，双萤光素酶报告基因系统检测miR-139-5p与CXCR4之间的靶向性，实时荧光定量聚合酶链式反应(Real-time quantitative polymerase chain reaction，Real-time qPCR)检测miR-139-5p表达，免疫印迹法检测CXCR4蛋白表达。[结果] 与正常对照组比较，转染miR-139-5p mimic后，细胞划痕愈合减慢(P<0.01)，Transwell小室迁移的细胞数量减少(P<0.01)，CXCR4蛋白表达下调(P<0.05)，而转染miR-139-5p inhibitor的作用相反。双萤光素酶报告基因检测结果表明，CXCR4是miR-139-5p的潜在靶基因(P<0.01)。与正常对照组比较，50、100 μmol·L-1 TMP预处理均下调miR-139-5p表达(P<0.01)；与TMP组比较，TMP+miR-139-5p mimic组划痕愈合减慢(P<0.01)，Transwell 小室迁移的细胞数量减少(P<0.01)，CXCR4蛋白表达下调(P<0.01)，TMP+miR-139-5p mimic NC组无明显变化(P>0.05)。[结论] TMP可促进BMSCs迁移，其机制可能与下调miR-139-5p，增加CXCR4的表达有关。 |
| 英文摘要: |
| [Objective] To investigate whether tetramethylpyrazine(TMP) regulates the migration of bone marrow mesenchymal stem cells(BMSCs) through miR-139-5p/chemokine receptor 4(CXCR4). [Methods] BMSCs from SD rats were cultured by whole bone marrow adherent method. BMSCs were pretreated with 50，100 μmol·L-1 TMP for 24 h， then miR-139-5p mimic/inhibitors were transfected using lipofectamine 3000. Cell migration ability was detected by cell scratch and Transwell test. Dual luciferase reporter system was used to detect the targeting between miR-139-5p and CXCR4. The miR-139-5p expression was tested by Real-time quantitative polymerase chain reaction(Real-time qPCR). The CXCR4 protein expression was measured by Western blot. [Results] Compared with normal control group， the cell scratch healing was slown(P<0.01)， the number of cells crossing the Transwell chamber was decreased(P<0.01)， and CXCR4 protein expression was down-regulated(P<0.05) after transfection with miR-139-5p mimic， but miR-139-5p inhibitor had the opposite effects. Dual luciferase reporter gene result indicated that CXCR4 was a potential target gene of miR-139-5p(P<0.01). Compared with normal control group， miR-139-5p expression was down-regulated after 50，100 μmol·L-1 TMP treatment(P<0.01). Compared with TMP group， the cell scratch healing was slown(P<0.01)， the number of cells crossing the Transwell chamber was decreased(P<0.01)， and CXCR4 protein expression was down-regulated(P<0.01) in TMP+miR-139-5p mimic group， but there were no significant changes in TMP+miR-139-5p mimic NC group(P>0.05). [Conclusion] TMP can promote the migration capacity of BMSCs， the mechanism may be related to downregulating miR-139-5p， and increasing the expression of CXCR4. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |