Wnt信号通路对骨髓间充质干细胞CXCR4表达的调控作用  

The effects of canonical Wnt signaling pathways on the CXCR4 expression of bone marrow derived mesenchymal stem cells

作　　者：孙吉平[1] 马莹 张文静[1] 吕佳[1] 尹爱萍[1] 

机构地区：[1]西安交通大学医学院第一附属医院肾内科,西安710061

出　　处：《临床肾脏病杂志》2017年第12期751-756,共6页Journal Of Clinical Nephrology

基　　金：国家自然科学基金资助项目(No.81200528)

摘　　要：目的研究经典Wnt信号通路对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)CXCR4表达的影响,探索Wnt通路调节干细胞迁移的可能机制。方法培养雄性SD大鼠BMSCs,对所培养的细胞进行鉴定;通过反转录-聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)探讨活化/阻断Wnt信号通路对BMSCs表达CXCR4的影响。结果(1)BMSCs的体外培养及鉴定:细胞呈梭形,漩涡状排列,放射状生长。细胞表面标志CD29(+)、CD90(+)、CD45(–),能定向诱导细胞表达神经细胞标记物,体外能分化为脂肪细胞,符合BMSCs表面标记物表达及多向分化潜能特征。(2)应用200 ng/ml的Wnt3a作用24 h即可以充分激活经典Wnt信号通路,应用100 ng/ml的DKK-1作用24 h可以充分阻断经典Wnt信号通路。(3)阻断经典Wnt信号通路可以促进体外培养的BMSCs的CXCR4的表达;激活经典Wnt信号通路可以抑制体外培养的BMSCs的CXCR4的表达。结论干预Wnt信号通路可以调控BMSCs CXCR4表达,经典Wnt信号通路有望通过CXCR4/SDF-1通路调控BMSCs向组织的迁移。Objective To evaluate the effects of Wnt signaling pathways on the CXCR4 expression of bone marrow mesenchymal stem cells（BMSCs）,and to investigate the possible regulation mechanisms of the Wnt signaling pathways on the migration ability of BMSCs.Methods BMSCs were obtained from the femurs,tibias,humeri and radius of adult male SD rats.The expression of Wnt signaling pathway downstream target geneβ-catenin was detected via RT-PCR and western-blotting after activating and blocking Wnt signaling pathway of BMSs.Meanwhile,expression of CXCR4 was also detected via RT-PCR and western-blotting after activating and blocking Wnt signaling pathway of BMSCs.Results（1）Dentification of BMSCs：BMSCs could expression CD29 and CD90,which were both marker of mesenchymal stem cells.Besides,BMSCs could express nerve culluar marker such as NSE,GFAP,and could be induced to differentiate into adipocytic cells,suggesting multi-directional differentiation.（2）Wnt3 a（200 ng/ml）could fully activate Wnt signaling pathway in 24 h,with high expression level ofβ-catenin（compared with control group,P〈0.05）.While DKK-1（100 ng/ml）could fully blockade Wnt signaling pathway in24 h（compared with control group,P〈0.05）.（3）When the Wnt signaling pathway was blockaded,expression levels of CXCR4 in BMSCs was obviously increased（compared with control group,P〈0.05）;while the activated of such signaling pathway could decrease the expression of CXCR4 of BMSCs（compared with control group,P〈0.05）.Conclusions Wnt signaling pathway could regulate the expression of CXCR4 of BMSCs,and such regulation maypromote the migration of BMSCs to kidney.

关 键 词：骨髓间充质干细胞 经典Wnt信号通路 Β-CATENIN CXCR4 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]