毛冬青甲素促进骨髓间充质干细胞迁移的实验研究  

Ilexonin A Promoted the Migration of Bone Mesenchymal Stem Cells

作　　者：郑关毅[1] 李庆双 张碧琴 陈晓东[4] 江琼[4] ZHENG Guan-yi;LI Qing-shuang;ZHANG Bi-qin;CHEN Xiao-dong;JIANG Qiong(Department of Traditional Chinese Medicine,Fujian Medical University Union Hospital,Fuzhou（350001;Department of Geriatrics,174th Hospital of the People＇s Liberation Army,Xiamen,Fujian（361000;Department of Geriatrics,First Hospital of Fuzhou,Fuzhou（350009;Institute of Burn,Fujian Medical University Union Hospital,Fuzhou（350001)

机构地区：[1]福建医科大学附属协和医院中医科,福州350001 [2]中国人民解放军第一七四医院老年科,厦门361000 [3]福州市第一医院老年科,福州350009 [4]福建医科大学附属协和医院烧伤研究所,福州350001

出　　处：《中国中西医结合杂志》2018年第11期1379-1383,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：福建省自然科学基金科技资助项目(No.2014J01327)

摘　　要：目的观察毛冬青甲素(ilexonin A, IA)干预大鼠骨髓间充质干细胞(BMSCs)后基质细胞衍生因子(SDF-1)特异性受体CXCR4和Wnt通路中β-连环蛋白(β-catenin)、糖原合成激酶(GSK-3β)的表达情况,探讨IA预处理促进BMSCs体外迁移的分子机制。方法采用全骨髓贴壁法体外分离、培养、纯化大鼠并行BMSCs鉴定。将BMSCs分为对照组、Wnt3a组、IA组、IA+Wnt3a组、IA+Dkk1组、Dkk1组。以Wnt信号通路的激动剂Wnt3a、抑制剂Dkk1干预,Western blot法观察β-catenin、GSK-3β与CXCR4的表达关系。结果与对照组比较,Wnt3a组、IA组CXCR4、β-catenin蛋白表达显著增强、GSK-3β蛋白表达显著减弱;(P<0.05),Dkk1组CXCR4、β-catenin蛋白表达显著减弱(P<0.05);与Wnt3a组、IA组分别比较,IA+Wnt3a组CXCR4蛋白显著增强,GSK-3β蛋白表达显著减弱(P<0.05);与Dkk1组比较,IA+Dkk1组CXCR4、β-catenin蛋白表达显著增强,GSK-3β蛋白表达显著减弱(P<0.05);与IA组比较,IA+Dkk1组β-catenin的表达显著减弱,GSK-3β蛋白表达显著增强(P<0.05)。结论 IA上调CXCR4的表达,可能与BMSCs的Wnt/β-catenin信号通路激活有关,发挥其促迁移的作用。Objective To observe the expressions of chemokine CXC motif receptor4 （CXCR4）, β-catenin and glycogen synthesis kinase 3β （GSK-3β） of the Wnt pathway after ilexonin A （IA） intervened bone mesenchymal stem cells （BMSCs） in vitro, and to elucidate molecular mechanism of IA for enhancing in vitro migration ability of BMSCs. Methods By using whole bone marrow adherent method, isolation, cultivation, purification, and identification of rat BMSCs in vitro were performed. BM- SCs were divided into the blank control group, Wnt3a group, IA group, IA ＋ Wnt3a group, IA ＋ Dkkl group, Dkkl group. After intervened by activator Wnt3a and inhibitor Dkkl, the expression relations among β-catenin, GSK-3β, and CXCR4 were observed by Western blot. Results Compared with the control group, expressions of CXCR4 and β-catenin protein were significantly increased in Wnt3a group and IA group （P 〈0.05）, GSK-3β protein expression was significantly attenuated in Dkkl group （P 〈0.05） CXCR4 and β-catenin pretein expressions were significantly attenuated in Dkkl group （P 〈0.05）. Compared with Wnt3a group and IA group respectively, expression of CXCR4 was significantly increased in IA ＋Wnt3a group （P 〈0.05）, GSK-313 protein expression was significantly attenuated （P 〈0.05）. Compared with Dkkl group, expressions of CXCR4 and β-catenin were significantly increased, GSK-3β protein expression was significantly attenuated （P 〈0.05） in IA ＋ Dkk1 group （P 〈0.05）. Compared with IA group, expression of β-catenin was significantly attenuated, and GSK-3β protein expression was significantly enhanced （P 〈0.05） in IA ＋Dkk1 group （P 〈0.05）. Conclusion IA might up-regulate the expression of CXCR4, which might be related with activating Wnt/β-catenin signaling pathway of BMSCs and playing its role in promoting migration.

关 键 词：骨髓间充质干细胞 毛冬青甲素 CXC趋化因子受体蛋白4 Wnt/β-连环蛋白 糖原合成激酶3β 

分 类 号：R285[医药卫生—中药学]