中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (23): 4291-4294.doi: 10.3969/j.issn.1673-8225.2011.23.025

• 干细胞与中医药 stem cells and traditional Chinese medicine • 上一篇    下一篇

独参汤促进骨髓间充质干细胞体外增殖：最佳干预剂量的筛选

王艳春1，2，闫承慧2，刘  晶2，韩雅玲2   

  1. 1辽宁中医药大学附属医院，辽宁省沈阳市 110032
    2解放军沈阳军区总医院，辽宁省沈阳市 110016
  • 收稿日期:2010-12-23 修回日期:2011-03-08 出版日期:2011-06-04 发布日期:2011-06-04
  • 通讯作者: 韩雅玲，博士，教授，主任医师，解放军沈阳军区总医院，辽宁省沈阳市 110016
  • 作者简介:王艳春☆，男，1974年生，辽宁省沈阳市人，汉族，2011年辽宁中医药大学毕业，博士，主治医师，主要从事中西医结合心血管病专业方面的研究。 w20090807@qq.com

Dushentang promotes the proliferation of bone marrow mesenchymal stem cells in vitro: Screening of the optimal intervention concentration  

Wang Yan-chun1,2, Yan Cheng-hui2, Liu Jing2, Han Ya-ling2   

  1. 1Affiliated Hospital, Liaoning University of Traditional Chinese Medicine, Shenyang  110032, Liaoning Province, China
    2General Hospital of Shenyang Military Area Command, Shenyang  110840, Liaoning Province, China
  • Received:2010-12-23 Revised:2011-03-08 Online:2011-06-04 Published:2011-06-04
  • Contact: Han Ya-ling, Doctor, Professor, Chief physician, General Hospital of Shenyang Military Area Command, Shenyang 110840, Liaoning Province, China
  • About author:Wang Yan-chun☆, Doctor, Attending physician, Affiliated Hospital, Liaoning University of Traditional Chinese Medicine, Shenyang 110032, Liaoning Province, China; General Hospital of Shenyang Military Area Command, Shenyang 110016, Liaoning Province, China w20090807@qq.com

PDF

418

摘要：

背景：骨髓间充质干细胞移植后极低的生存率限制了它的治疗作用，现正在寻找合适的药物促进其增殖。
目的：观察独参汤是否能促进骨髓间充质干细胞的增殖，并筛选出最佳干预浓度。
方法：分离培养Wistar大鼠骨髓间充质干细胞。培养、传代至第5代，利用流式细胞仪检测表面分子标记物。以每孔1×104数量种于96孔板。将独参汤颗粒剂分别以10，5，2.5，1.25，…0.019 5 g/L的质量浓度干预骨髓间充质干细胞，于第1，7天MTT法检测其增殖情况。
结果与结论：①原代细胞接种2 d后多数细胞贴壁，传代后细胞贴壁速度和增殖更快，第5代基本纯化为骨髓间充质干细胞，细胞呈放射状或漩涡状排列。②第5代骨髓间充质干细胞CD44阳性细胞占94.7%，CD90阳性细胞占86.8%，CD34阴性细胞占98.7%，CD45阴性细胞占97.1%。③独参汤浓度在0.625，0.312 5，0.156，0.078，0.039，0.019 5 g/L质量浓度时均可以促进骨髓间充质干细胞在体外的增殖。其中在质量浓度为0.078 g/L时作用最强。

关键词: 独参汤, 骨髓间充质干细胞, 增殖, 最佳浓度, 中药

Abstract:

BACKGROUND: The low survival rate of transplanted bone marrow mesenchymal stem cells (BMSCs) limits their therapeutic action. It is reasonable for us to look for a suitable drug to promote it proliferation.
OBJECTIVE: To investigate whether Dushentang may enhance the proliferation of BMSCs, and to screen the optimal treating concentration.
METHODS: After Wistar rats were anesthetized, BMSCs were obtained from the femoral and tibial bones, and then cultured to the fifth passage. Surface molecule markers of BMSCs were examined after 5 passages by flow cytometry. Cells were seeded in a 96-well microtiter plate at a density of 1×104 cells/well. Cells were then treated with Dushentang at concentration 10, 5, 2.5, 1.25… and 0.019 5 g/L for 1 or 7 days. Proliferation was determined after Dushentang treatment using colorimetry based on the uptake of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide salt (MTT) by viable cells.
RESULTS AND CONCLUSION: ①Most primary BMSCs adhered to the wall at 2 days after culture, which proliferated faster after passaged, and the fifth passage of cells were mostly purified into BMSCs, spread radially or vortex-likely. ②Surface molecule markers of the fifth passage of BMSCs were examined. The isolated cell purities were 86.8% for CD90, 94.7% for CD44, but negative for CD34 (98.7%) and CD45 (97.1%). ③Proliferation of BMSCs might be enhanced by Dushentang treatment with the concentration from 0.625, 0.312 5, 0.156, 0.078, 0.039, and 0.019 5 g/L, and the optimal concentration was 0.078 g/L.

中图分类号: