中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (11): 1915-1918.doi: 10.3969/j.issn.1673-8225.2012.11.004

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

地塞米松可影响兔髓核细胞增殖及Ⅱ型胶原的表达★

周小锐，刘世清，贺  斌，张  弩   

  1. 武汉大学人民医院骨三科，湖北省武汉市  430060
  • 收稿日期:2011-12-25 修回日期:2012-02-13 出版日期:2012-03-11 发布日期:2012-03-11
  • 作者简介:周小锐★，男，1983年生，湖北省十堰市人，汉族，武汉大学第一临床学院在读硕士，医师，主要从事脊柱脊髓损伤的研究。 isaiah605@sohu.com

Dexamethasone effects on the proliferation of nucleus pulposus cells and the expression of type Ⅱ collagen-2 in rabbits

Zhou Xiao-rui, Liu Shi-qing, He Bin, Zhang Nu   

  1. Third Department of Orthopaedics, Renmin Hospital of Wuhan University, Wuhan  430060, Hubei Province, China
  • Received:2011-12-25 Revised:2012-02-13 Online:2012-03-11 Published:2012-03-11
  • About author:Zhou Xiao-rui★, Studying for master’s degree, Physician, Third Department of Orthopaedics, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China isaiah605@sohu.com

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摘要：

背景：地塞米松是细胞增殖的一种常用药物，但能否促进髓核细胞增殖呢？
目的：观察地塞米松对兔椎间盘髓核细胞增殖的影响，以及对髓核细胞分泌Ⅱ型胶原的影响。
方法：无菌条件下取兔椎间盘，常规分离消化髓核细胞进行培养；传代培养第2代髓核细胞7 d，加入塞米松浓度分别为1，10，100，1 000，10 000 nmol/L，培养1~5 d每日MTT常规测一组490 nm处的吸光值并制定生长曲线。另选取最佳作用浓度100 nmol/L地塞米松加入髓核细胞进行培养，以不加于地塞米松为对照。
结果与结论：MTT检测结果表明地塞米松对兔髓核细胞增殖有促进作用，最佳作用浓度为100 nmol/L，最佳作用时间为    48 h；RT-PCR检测结果表明经地塞米松处理的髓核细胞其Ⅱ型胶原表达明显较对照组高(P < 0.05)。提示地塞米松可以促进兔椎间盘髓核细胞的增殖，并可使兔髓核细胞内Ⅱ型胶原表达增高。

关键词: 地塞米松, Ⅱ型胶原, 髓核细胞, 兔, 组织工程

Abstract:

BACKGROUND: Dexamethasone is a common drug for cell proliferation. However, it remains unclear whether dexamethasone can promote the proliferation of nucleus pulposus cells.
OBJECTIVE: To investigate the effects of dexamethasone on the proliferation of nucleus pulposus cells in rabbit intervertebral disk and the expression of type Ⅱ collagen secreted by nucleus pulposus cells.
METHODS: The rabbit intervertebral disk was extracted in sterile processing. After conventional separation and digestion, nucleus pulposus cells were cultured and passaged for 7 days, and then dexamethasone with the concentration of 1, 10, 100,    1 000, 10 000 nmol/L was added. After that, the nucleus pulposus cells were cultured for 1-5 days, and the absorbance values at 490 nm of the cells were detected by 3-(4, 5)-dimethylthiahiazo (-z-y1)-3, 5-di-phenytetrazoliumromide (MTT) assay every day and the growth curve was drawn. In addition, 100 nmol/L dexamethasone was selected for culture of nucleus pulposus cells, and the nucleus pulposus cells cultured with no dexamethasone was as controls.
RESULTS AND CONCLUSION: MTT assay results showed that dexamethasone had the proliferative effects on the cultured nucleus pulposus cells. The optimal action time was 48 hours and the optimal action concentration was 100 nmol/L. Reverse transcription-PCR results showed that dexamethasone could up-regulate the mRNA expression of type Ⅱ collagen which was significantly higher than that in the control group (P < 0.05). These suggest that dexamethasone can promote the proliferation of nucleus pulposus cells of rabbit intervertebral disk and up-regulate the expression of type Ⅱ collagen in rabbit nucleus pulposus cells.

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