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

作　　者：梁慧琦 李琳[1] 杨琰[1] 许家栋[1] 徐岚溪 潘小平 储利胜[1] 方燕[1] LIANG Huiqi;LI Lin;YANG Yan(School of Basic Medical Sciences,Zhejiang Chinese Medical University,Hangzhou(310053),China)

机构地区：[1]浙江中医药大学基础医学院,杭州310053

出　　处：《浙江中医药大学学报》2023年第7期703-711,共9页Journal of Zhejiang Chinese Medical University

基　　金：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：[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、100μmol·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、100μmol·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。[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 50,100μmol·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 50,100μmol·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.

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 

分 类 号：R285.5[医药卫生—中药学]