毛冬青甲素对大鼠骨髓间充质干细胞增殖及迁移的影响  

Effect of ilexonin A on proliferation and migration of bone mesenchymal stem cells in rats

作　　者：李庆双 郑关毅[1] 张碧琴 陈晓东[4] 江琼[4] 刘真臻 

机构地区：[1]福建医科大学附属协和医院中医科,福建福州350001 [2]中国人民解放军第一七四医院老年科,福建厦门361000 [3]福州市第一医院老年科,福建福州350009 [4]福建医科大学附属协和医院烧伤研究所,福建福州350001 [5]山东省临沂市人民医院老年科,山东临沂276000

出　　处：《中国药理学通报》2018年第3期358-364,共7页Chinese Pharmacological Bulletin

基　　金：福建省自然科学基金资助项目(No 2014J01327)

摘　　要：目的观察毛冬青甲素(ilexonin A,IA)对大鼠骨髓间充质干细胞(BMSCs)增殖的影响,并探讨IA是否通过上调CXCR4的表达促进大鼠BMSCs迁移。方法采用MTT比色法检测IA(3.125、6.25、12.5、25、50、100、200、400、800 mg·L^(-1))预处理BMSCs 24、48、72 h,观察对BMSCs增殖的影响,确定最佳药物浓度和最佳作用时间。用最佳浓度的IA干预第3代BMSCs 48 h,Transwell检测BMSCs迁移能力,Western blot法检测CXCR4的表达水平。结果 MTT结果显示,IA孵育BMSCs 24、48 h,与对照组相比,100~800 mg·L^(-1)组的细胞增殖明显减少(P<0.05);IA孵育BMSCs 48h,与对照组相比,6.25、3.125 mg·L^(-1)组细胞增殖明显增加(P<0.05);IA孵育BMSCs 72 h,与对照组相比,12.5~800mg·L^(-1)组细胞增殖明显减少(P<0.05),提示IA 6.25、3.125 mg·L^(-1)剂量组于48 h预处理BMSCs是最优选择。Transwell细胞迁移实验表明,IA(6.25、3.125 mg·L^(-1))预处理BMSCs 48 h明显增强BMSCs运动迁移能力(P<0.05),且迁移与IA浓度无关,CXCR4拮抗剂AMD3100可完全阻断其促迁移效应。Western blot显示,IA(6.25、3.125 mg·L^(-1))预处理BMSCs 48 h,上调CXCR4蛋白表达(P<0.05)。结论 IA可以促进BMSCs的增殖,并通过上调CXCR4的表达,提高BMSCs的迁移能力。bAim To observe the effect of ilexonin A（ IA） on the proliferation of bone marrow mesenchymal stem cells（ BMSCs）,and to investigate whether IA can promote the migration of BMSCs by up-regulating the expression of CXCR4 in rats. Methods MTT method was used to assay and analyse the proliferation of BMSCs which were pretreated with different concentrations of IA（ 3. 125,6. 25,12. 5,25,50,100,200,400,800 mg·L^（-1）） for 24,48 and 72 h,then the best concentration and the best optimum time were screened.The third generation of BMSCs was exposed to the optimal concentration of IA for 48 h. The Transwell system was used to carry out the experiment of BMSCs migration. Western blot was used to analyse the expression of CXCR4. Results MTT assay showed that compared with control group,the proliferation of BMSCs was significantly reduced in IA 100 ~ 800 mg · L^（-1） groups at 24 h（ P〈 0. 05）; compared with control group, the proliferation of BMSCs significantly decreased in IA 100 ~ 800 mg·L^（-1） groups at 48 h（ P 0. 05）,but markedly increased in IA 6. 25 and 3. 125 mg·L^（-1） groups（ P 〈0. 05）; compared with control group,the proliferation of BMSCs was significantly reduced in IA 12. 5 ~ 800 mg·L^（-1） groups at 72 h（ P 〈0. 05）. The above results indicated that the BMSCs incubated with IA 6. 25 and 3. 125 mg · L^（-1） for 48 h were the optimal choice to promote proliferation. The Transwell migration assay showed that incubation with IA 6. 25 and 3. 125 mg·L^（-1） for 48 h could significantly increase the migration of BMSCs（ P 〈0. 05）,and the migration rate was not related with the concentration of IA. This effect was completely blocked by AMD3100（ the antagonist of CXCR4）. Western blot showed that incubation with IA 6. 25 and 3. 125 mg·L^（-1） for 48 h could increase the expression of CXCR4 in BMSCs（ P 〈0. 05）. Conclusion IA can promote the proliferation of BMSCs and increase the migration of BMSCs by up-regulating the expression of CXCR4.

关 键 词：骨髓间充质干细胞 迁移 毛冬青甲素 CXCR4 增殖 孵育 

分 类 号：R-332[医药卫生]