中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (14): 2564-2567.doi: 10.3969/j.issn.1673-8225.2010.14.019

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

黄芪注射液对体外培养嗅鞘细胞增殖及其分泌神经营养因子的影响

王培民1，李剑锋2，张农山3，张  旭3   

  1. 1江苏省中医院，江苏省南京市 210029；2内蒙古医学院第二附属医院，内蒙古自治区呼和浩特市 010030；3南京中医药大学，江苏省南京市 210029
  • 出版日期:2010-04-02 发布日期:2010-04-02
  • 通讯作者: 张 旭，博士生导师，南京中医药大学，江苏省南京市 210029 zhangxutcm@gmail.com
  • 作者简介: 王培民☆，男，1963年生，山东省文登市人，汉族，2005年南京中医药大学毕业，博士，主任医师，主要从事创伤与骨关节炎的临床与实验研究。 drwpm@163.com
  • 基金资助:

    国家科技支撑计划“中医药诊疗技术及特色产品应用研究”重点项目(2006BAI11B08)：中药复方及有效成分的系统生物学评价。

Influence of astragalus injection on in vitro cultured olfactory ensheathing cell proliferation and neurotrophic factor secretion

Wang Pei-min1, Li Jian-feng2, Zhang Nong-shan3, Zhang Xu3   

  1. 1 Jiangsu Provincial Hospital of Traditional Chinese Medicine, Nanjing   210029, Jiangsu Province, China; 2 Second Hospital of Inner Mongolia Medical College, Hohhot   010030, Inner Mongolia Autonomous Region, China;  3 Nanjing University of Chinese Medicine, Nanjing   210029, Jiangsu Province, China
  • Online:2010-04-02 Published:2010-04-02
  • Contact: Zhang Xu, Doctoral supervisor, Nanjing University of Chinese Medicine, Nanjing 210029, Jiangsu Province, China zhangxutcm@gmail.com
  • About author:Wang Pei-min☆, Doctor, Chief physician, Jiangsu Provincial Hospital of Traditional Chinese Medicine, Nanjing 210029, Jiangsu Province, China drwpm@163.com
  • Supported by:

     the National Science and Technology Support Program, No. 2006BAI11B08*

PDF

376

被引次数

20

摘要：

背景：单独黄芪注射或嗅鞘细胞移植均可促进多种神经元的存活及脊髓损伤后神经功能的恢复，若联合嗅鞘细胞移植和中药黄芪注射液治疗脊髓损伤，是否可达到更好临床效果？
目的：观察黄芪注射液对嗅鞘细胞增殖及其分泌神经营养因子的影响。
方法：分离培养新生24 h内SD大鼠嗅鞘细胞并纯化，采用免疫组织化学方法鉴定。取培养至第8天的嗅鞘细胞，种植于多聚赖氨酸包被的96孔板，置于37 ℃、体积分数5%CO2培养箱饥饿24 h后，加入2 mg/L，20 mg/L，200 mg/L，2 g/L，20 g/L黄芪注射液作用24 h，采用MTT法和流式细胞仪检测黄芪注射液对嗅鞘细胞增殖的影响；采用ELISA法测定培养上清中胶质细胞源性神经营养因子的水平。以血清培养液组为阴性对照。
结果与结论：与阴性对照组比较，2，20 mg/L质量浓度黄芪注射液可明显促进嗅鞘细胞增殖(P < 0.05，P < 0.01)；不同质量浓度黄芪注射液均增加嗅鞘细胞G1细胞比例，减少S、G2期细胞所占比例，但差异无显著性意义(P > 0.05)，但各质量浓度黄芪注射液均显著减少嗅鞘细胞凋亡数量(P < 0.05)； 2 mg/L质量浓度黄芪能对嗅鞘细胞分泌胶质细胞源性神经营养因子有显著的促进作用。结果说明黄芪注射液可协同嗅鞘细胞促进脊髓损伤的恢复。

关键词: 黄芪, 嗅鞘细胞, 增殖, 神经营养因子, 干细胞, 神经干细胞

Abstract:

BACKGROUND: Astragalus or olfactory ensheathing cell (OECs) transplantation alone can promote survival of various neurons and functional improvement of injured spinal cord. The clinical effect of astragalus in combination with OECs transplantation remains unclear.
OBJECTIVE: To observe the effect of astragalus on the proliferation of OECs and the secretion of neurotrophic factors. 
METHODS: OECs of SD rats, 24 hours old, were isolated and cultured, and identified by immunohistochemistry. OECs at culture day 8 were seeded in polylysine-coated 96-well plate and incubated in 5% CO2 at 37 ℃ for 24 hours, followed by   2 mg/L, 20 mg/L, 200 mg/L, 2 g/L, 20 g/L astragalus injection for 24 hours. MTT and flow cytometry were used to assess the effects of astragalus in promoting OECs proliferation; enzyme linked immunoassay (ELISA) was used to measure the content of neurotrophic factors. Serum culture medium was served as negative control. 
RESULTS AND METHODS: Compared with the negative control group, 2 and 20 mg/L astragalus injection significantly promoted OECs proliferation (P < 0.05, P < 0.01). The result of flow cytometry showed that astragalus could promote OECs in G1 stage and reduce the percentage of cells in S and G2 stages, but the difference was not significant (P > 0.05). However, astragalus injection at each mass concentration significantly reduced the number of OECs (P < 0.05). Moreover, 2 mg/L astragalus injection significantly promoted in vitro neurotrophic factor levels in OECs. Results show that astragalus injection in combination with OECs can promote recovery of spinal cord injury.

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