[1]吴志方 罗毅文△ 王斌 蔡炎 罗辉 黄杰文 刘亚梅 黄永铨.补肾活血汤石油醚提取物对骨髓间充质干细胞表达Wnt5a/CCR2的影响[J].中国中医骨伤科杂志,2017,25(07):6-10，15.
　WU Zhifang LUO Yiwen WANG Bin CAI Yan LUO Hui HUANG Jiewen LIU Yamei HUANG Yongquan.Effect of Petroleum Ether Extracts from Bushenhuoxue Decoction on the Expression of Wnt5a/CCR2 in Bone Marrow Mesenchymal Stem Cells[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2017,25(07):6-10，15.
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补肾活血汤石油醚提取物对骨髓间充质干细胞表达Wnt5a/CCR2的影响()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第25卷
期数:
2017年07期
页码:
6-10，15
栏目:
实验研究
出版日期:
2017-07-15

文章信息/Info

Title:
Effect of Petroleum Ether Extracts from Bushenhuoxue Decoction on the Expression of Wnt5a/CCR2 in Bone Marrow Mesenchymal Stem Cells
文章编号:
1005-0205(2017)07-0006-05
作者:
吴志方1 罗毅文1△ 王斌1 蔡炎1 罗辉1 黄杰文1 刘亚梅2 黄永铨3
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系
3.广东省中医院
△.通信作者 E-mail:gzhluoyw@126.com
Author(s):
WU Zhifang1 LUO Yiwen1 WANG Bin1 CAI Yan1 LUO Hui1 HUANG Jiewen1 LIU Yamei2 HUANG Yongquan3
1.Otrhopedics and Traumatology Hospital, Affiliated to Guangzhou University of Traditional Chinese Medicine, Guangzhou 510240, China;
2.Department of Traditional Chinese Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510006, China;
3.Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou 510120, China.
关键词:
补肾活血汤;  骨髓间充质干细胞;  体外迁移;  信号通路;  C-C类趋化因子受体2
Keywords:
Bushenhuoxue decoction;  bone marrow mesenchymal stem cells;  migration in vitro;  signaling pathway;  C-C chemokine receptor 2
分类号:
R-33
文献标志码:
A
摘要:
目的:探讨补肾活血汤石油醚提取物对骨髓间充质干细胞(BMSCs)表达CCR2及其Wnt5a的影响。方法:索氏提取法提取补肾活血汤石油醚提取物; 全骨髓贴壁筛选法培养BMSCs,取BMSCs第三代用于实验。石油醚部位按10,50,100 μg/mL浓度梯度刺激BMSCs,利用免疫蛋白印迹法(Western blot)及其细胞免疫荧光检测CCR2蛋白表达情况,采用Western blot和qPCR检测Wnt5a蛋白和mRNA的表达。结果:Western blot和细胞免疫荧光结果显示石油醚提取物按照浓度梯度促进CCR2蛋白表达,以100 μg/mL浓度最佳(P<0.01)。石油醚提取物同样促进Wnt5a蛋白及mRNA表达,其中Western blot结果亦以100 μg/mL浓度最佳(P<0.01)。结论:补肾活血汤石油醚提取物亦能刺激MCP-1/CCR2信号轴,上调Wnt5a蛋白表达,初步发现迁移活性可能与Wnt5a通路相关。
Abstract:
Objective: To investigate the effect of petroleum ether extracts from Bushenhuoxue decoction(BSHXD)on the expression of CCR2 and Wnt5a in bone marrow mesenchymal stem cells(BMSCs). Methods: Soxhlet extraction method was used to extract petroleum ether extracts from BSHXD. BMSCs were isolated and cultured by whole bone marrow adherent method, and the third generation of BMSCs were used for the experiment. Petroleum ether extracts was used to stimulate BMSCs at the concentration gradient of 10, 50 and 100 μg/mL. The expression of CCR2 protein was detected by Western blot and cell immunofluorescence, and the expression of Wnt5a protein and mRNA was detected by Western blot and qPCR. Results:The results of Western blot and cell immunofluorescence showed that petroleum ether extracts up-regulated the expression of CCR2 protein in a dose-dependent manner and the active concentration was 100 μg/mL(P<0.01). And petroleum ether extracts also up-regulated the expression of Wnt5a protein and mRNA, the active concentration in Western blot was also 100 μg/mL(P<0.01). Conclusion:Petroleum ether extracts from BSHXD can also stimulate the MCP-1/CCR2 signaling axis and upregulate the expression of Wnt5a protein, it preliminarily suggests that the migration activity may be related to Wnt5a signaling pathway.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金(81473699,81273783)
更新日期/Last Update: 2017-07-15