中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (10): 1817-1820.doi: 10.3969/j.issn.1673-8225.2010.10.022

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

黄芪甲甙对大鼠骨髓间充质干细胞多种造血相关因子表达的影响

谭艳芳1，殷小成1，熊玉娟2，王 艳1   

  1. 1南华大学附属第一医院儿科，湖南省衡阳市 421001；
    2湖南省儿童医院康复科，湖南省长沙市 410000
  • 出版日期:2010-03-05 发布日期:2010-03-05
  • 通讯作者: 殷小成，硕士，副主任医师，硕士生导师，南华大学附属第一医院儿科，湖南省衡阳市 421001 xcyin108@sina. com
  • 作者简介:谭艳芳，女，1981年生，湖南省长沙市人，汉族，南华大学附属第一医院在读硕士，医师，主要从事儿童白血病及其他恶性血液系统疾病方面的研究。 23289616@qq. com

Effect of Astragaloside IV on hematopoietic growth factor expression in rat bone marrow mesenchymal stem cells

Tan Yan-fang1, Yin Xiao-cheng1, Xiong Yu-juan2, Wang Yan1   

  1. 1Department of Pediatrics, First Hospital Affiliated to University of South China, Hengyang   421001, Hunan Province, China;
    2Department of Rehabilitation, Hunan Children’s Hospital, Changsha   410000, Hunan Province, China
  • Online:2010-03-05 Published:2010-03-05
  • Contact: Yin Xiao-cheng, Master, Associate chief physician, Master’s supervisor, Department of Pediatrics, First Hospital Affiliated to University of South China, Hengyang 421001, Hunan Province, China xcyin108@sina.com
  • About author:Tan Yan-fang, Studying for master’s degree, Physician, Department of Pediatrics, First Hospital Affiliated to University of South China, Hengyang 421001, Hunan Province, China 23289616@qq.com

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486

被引次数

31

摘要：

背景：黄芪甲甙是中药黄芪的主要有效成分，能促进骨髓间充质干细胞的增殖及诱导分化，但涉及具体作用机制的报道较少。

目的：探讨中药黄芪甲甙对大鼠骨髓间充质干细胞造血相关因子表达的影响。

方法：全骨髓贴壁法和单克隆培养法体外分离培养大鼠骨髓间充质干细胞，接种于96孔板中，加药组分别用25，50，100，200 g/L黄芪甲甙100 μL孵育72 h，对照组仅加等量DMEM-LG培养液。间接免疫荧光法鉴定其生物学特性，MTT法检测黄芪甲甙对骨髓间充质干细胞增殖的影响，RT-PCR法检测黄芪甲甙干预后骨髓间充质干细胞造血相关因子的表达。

结果与结论：第3代骨髓间充质干细胞高表达CD44，低表达造血细胞表面标志CD45。与对照组比较，黄芪甲甙呈时间及剂量依赖性促进骨髓间充质干细胞的增殖，以200 g/L黄芪甲甙干预72 h促增殖作用最强(P < 0.05)。与对照组比较，加药组骨髓间充质干细胞SCF的表达显著增加(P < 0.01)，TPO，GM-CSF，TGF-β1等细胞因子的表达无明显变化(P > 0.05)，不表达白细胞介素3。黄芪甲甙可促进骨髓间充质干细胞的体外增殖，可能与其诱导骨髓间充质干细胞分泌SCF有关。

关键词: 黄芪甲甙, 造血相关因子, SCF, 增殖, 骨髓间充质干细胞

Abstract:

BACKGROUND: Astragaloside IV is a major component of Huangqi, promoting proliferation and differentiation of bone marrow mesenchymal stem cells; however, the mechanism has been less reported yet.

OBJECTIVE: To explore the effect of Astragaloside IV on expression of multiple hematopoietic growth factors in bone marrow mesenchymal stem cells.

METHODS: Bone marrow mesenchymal stem cells were isolated from adult Wistar rats by using the method of adhesive culture and clone, and they were then plated on 96-well plate and separately incubated with 100 μL Astragaloside IV (25, 50, 100, 200 g/L) for 72 hours. The cells in the control group were cultured with an equal volume of DMEM-LG culture liuquid. Indirect immunofluorescence was used to detect the biological activity, MTT method was used to evaluate the effect of Astragaloside IV on proliferation and differentiation of bone marrow mesenchymal stem cells, and RT-PCR method was used to measure the expression of hematopoietic growth factors in bone marrow mesenchymal stem cells.

RESULTS AND CONCLUSION: The 3rd-passage bone marrow mesenchymal stem cells highly expressed CD44 but lowly expressed CD45. As compared with control group, Astragaloside IV promoted proliferation of bone marrow mesenchymal stem cells in a time/dosage-dependent manner, in particular, the 200 g/L Astragaloside IV and 72-hour intervention (P < 0.05). SCF expression was significantly increased in the drug group compared with control group (P < 0.01); however, TPO, GM-CSF, and TGF-β1 expressions were not changed significantly (P > 0.05). Moreover, interleukin-3 expression was not found in the bone marrow mesenchymal stem cells. Astragaloside IV promoted in vitro proliferation of bone marrow mesenchymal stem cells, possibly involving in SCF secretion.

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