[1]王斌 罗毅文 黄永铨 刘亚梅△.补肾活血汤促骨折端间充质干细胞体外 迁移及CCR2表达的研究[J].中国中医骨伤科杂志,2015,23(08):1-5.
　WANG Bin LUO Yiwen HUANG Yongquan LIU Yamei.Research of Bushenhuoxue Decoction on Promoting the Migration of Mesenchymal Stem Cells in the Fracture Site and Expression of CCR2[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2015,23(08):1-5.
点击复制

补肾活血汤促骨折端间充质干细胞体外 迁移及CCR2表达的研究
分享到：

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第23卷
期数:
2015年08期
页码:
1-5
栏目:
实验研究
出版日期:
2015-08-15

文章信息/Info

Title:
Research of Bushenhuoxue Decoction on Promoting the Migration of Mesenchymal Stem Cells in the Fracture Site and Expression of CCR2
文章编号:
1005-0205(2015)08-0001-05
作者:
王斌1 罗毅文1 黄永铨1 刘亚梅2△
1.广州中医药大学附属骨伤科医院(广州,510240)
2.广州中医药大学中医诊断学教研室
△.通信作者 E-mail:gzhlym@163.com
Author(s):
WANG Bin1 LUO Yiwen1 HUANG Yongquan1 LIU Yamei2△
1.Orthopedics and Traumatology Hospital,Affiliated to Guangzhou University of Traditional Chinese Medicine,Guangzhou 510240,China;
2.Department of Diagnosis of Traditional Chinese Medicine,Guangzhou University of Traditional Chinese Medicine,Guangzhou 510405,China.
关键词:
补肾活血汤;  间充质干细胞;  体外迁移;  CCR2
Keywords:
Bushenhuoxue decoction;  mesenchymal stem cells;  migration in vitro;  CCR2
分类号:
R-33
文献标志码:
A
摘要:
目的:探讨补肾活血汤对骨折端间充质干细胞(Mesenchymal Stem Cells,MSCs)体外迁移及表面受体CC类趋化因子受体2(C-C motif chemokine receptor-2,CCR2)表达的影响。方法:补肾活血汤灌胃治疗大鼠股骨中段骨折模型7、14、21 d,分别取骨折端MSCs进行培养,传至第3代采用Transwell体外迁移实验检测补肾活血汤对骨折端MSCs体外迁移的影响,Western Blot、qPCR检测CCR2的表达。结果:补肾活血汤治疗后第7、14天,骨折端MSCs体外迁移能力明显加强,与模型组、对照组比较,差异有统计学意义(P<0.05)。第21天,骨折端MSCs体外迁移能力明显减弱,与模型组比较,差异无统计学意义(P>0.05)。Western blot、qPCR结果显示,补肾活血汤治疗第7天,治疗组CCR2的表达明显高于模型组、对照组(P<0.05)。第14天,CCR2的表达略有上升,与治疗后7 d比较,差异无统计学意义(P>0.05),但治疗组仍显著高于模型组、对照组(P<0.05)。第21天,CCR2的表达有所下降,与治疗后7 d比较,差异有统计学意义(P<0.05)。但治疗组与模型组比较,差异无统计学意义(P>0.05)。实验结果还表明,对照组CCR2的表达均处于低水平,7、14、21 d比较,差异无统计学意义(P>0.05)。结论:补肾活血汤可以促进大鼠骨折愈合过程中MSCs的体外迁移,其相关机制可能与上调CCR2的表达、激活MCP-1/CCR2信号轴有关。
Abstract:
Objective:To explore the effect of Bushenhuoxue decoction(BSHXD)on promoting the migration of mesenchymal stem cells(MSCs)in the fracture site and the expression of C-C motif chemokine receptor-2(CCR2).Methods:The rat femur fracture model was established and treated with BSHXD.Then the MSCs in the fracture site was isolated and cultured at 7th,14th,and 21st day,respectively.At the third passage,Transwell assay was applied to examine the migration of MSCs influenced by BSHXD.The expression of CCR2 was detected by Western blot and qPCR.Results:BSHXD could promote significantly more migration of MSCs at 7th and 14th day,but not at 21st day,compared with the model group and control group(P<0.05).The results of Western blot and qPCR showed that the expression of CCR2 in the treatment group at 7th day was significantly higher than the model group and control group(P<0.05).At 14th day,the expression of CCR2 increased slightly compared with 7th day,with no statistically significant(P>0.05),but it was still significantly higher than the model group and control group(P<0.05).At 21st day,the expression of CCR2 was declined significantly compared with the expression at 7th day(P<0.05),which was not statistically different between the treatment group and the model group(P>0.05).The results also show that the expression of CCR2 in the control group were at a low level,there was no significant difference among 7th,14th,and 21st day(P>0.05).Conclusion:BSHXD could promote migration of MSCs during fracture healing,and its mechanism may be associated with the up-regulation of CCR2 and activation of MCP-1/CCR2 signaling axis.

参考文献/References:

[1] Knight MN,Hankenson KD.Mesenchymal stem cells in bone regeneration[J].Advances in Wound Care,2013,2(6):306-316.
[2] Wu AC,Morrison NA,Kelly WL,et al.MCP-1 expression is specifically regulated during activation of skeletal repair and remodeling[J].Calcified Tissue International,2013,92(6):566-575.
[3] 曾意荣,樊粤光,刘红,等.补肾活血中药对大鼠骨髓间充质干细胞体外增殖的影响(英文)[J].中国组织工程研究与临床康复,2008,12(8):1581-1585.
[4] 张倍源.补肾活血方调控microRNA-210促进BMSCs成骨分化的研究[J].中国骨质疏松杂志,2014,20(4):396-399.
[5] Kang JW,Park KD,Choi Y,et al.Biodistribution and in vivo efficacy of genetically modified human mesenchymal stem cells systemically transplanted into a mouse bone fracture model[J].Arch Pharm Res,2013,36(8):1013-1022.
[6] 王斌,刘亚梅,罗毅文,等.SD大鼠皮质骨来源间充质干细胞的培养与鉴定[J].广州中医药大学学报,2014,31(3):452-455.
[7] Chamberlain G,Smith H,Rainger GE,et al.Mesenchymal stem cells exhibit firm adhesion,crawling,spreading and transmigration across aortic endothelial cells:effects of chemokines and shear[J].PloS One,2011,6(9):e25663.
[8] Ishikawa M,Ito H,Kitaori T,et al.MCP/CCR2 signaling is essential for recruitment of mesenchymal progenitor cells during the early phase of fracture healing[J].PloS One,2014,9(8):e104954.
[9] Goodman SB.Cell-based therapies for regenerating bone[J].Minerva Ortop Traumatol,2013,64(2):107-113.
[10] Choudhery MS,Badowski M,Muise A,et al.Donor age negatively impacts adipose tissue-derived mesenchymal stem cell expansion and differentiation[J].J Trans Med,2014,12(7):8.
[11] Gao C,Seuntjens J,Kaufman GN,et al.Mesenchymal stem cell transplantation to promote bone healing[J].J Orthop Res,2012,30(8):1183-1189.
[12] Ichida M,Yui Y,Yoshioka K,et al.Changes in cell migration of mesenchymal cells during osteogenic differentiation[J].FEBS Lett,2011,585(24):4018-4024.

备注/Memo

备注/Memo:
收稿日期:2014-10-08
基金项目:国家自然科学基金项目(81273783; 81473699)
更新日期/Last Update: 2015-08-15