《中国康复理论与实践》 ›› 2012, Vol. 18 ›› Issue (6): 539-543.

• 论文 • 上一篇    下一篇

碱性成纤维细胞生长因子对人椎间盘细胞外基质合成及软骨调节素表达的影响

李想1,2,王以朋3,洪毅1,2,唐和虎1,2,张军卫1,2,白金柱1,2,姜树东1,2,王方永1,2   

  1. 1.首都医科大学康复医学院,北京市100068;2.中国康复研究中心北京博爱医院脊柱脊髓外科,北京市100068;3.中国医学科学院,北京协和医学院,北京协和医院骨科,北京市100730。
  • 收稿日期:2012-02-27 修回日期:2012-05-21 出版日期:2012-06-25 发布日期:2012-06-25

Effect of Basic Fibroblast Growth Factor on Synthesis of Extracellular Matrixc and Expression of Chondromodulin in Human IntervertebralDisc Cells

LI Xiang, WANG Yi-peng, HONG Yi, et al.   

  1. Capital Medical University School of Rehabilitation Medicine, Departmentof Spine and Spinal Cord Surgery, Beijing Bo'ai Hospital, China Rehabilitation Research Center, Beijing 100068, China
  • Received:2012-02-27 Revised:2012-05-21 Published:2012-06-25 Online:2012-06-25

摘要: 目的研究碱性成纤维细胞生长因子(bFGF)对人椎间盘细胞基质合成能力以及血管生长抑制因子软骨调节素-1(ChM-1)表达的影响,为椎间盘退变的生物学治疗提供可供参考的生长因子。方法取4 例因腰椎间盘退变性疾病而于本院行腰椎间盘切除手术患者的椎间盘组织,分别进行髓核和纤维环细胞培养及表型鉴定。取传代细胞继续培养1 周后,向培养基中加入不同浓度的bFGF(0, 0.1 ng/ml, 1 ng/ml 和10 ng/ml),72 h 后收集细胞。采用Real-time RT-PCR 检测各组细胞中Aggrecan 和Ⅱ型胶原mRNA表达情况;同时利用Real-time RT-PCR和Western blot 方法检测bFGF 对ChM-1 mRNA和蛋白表达的影响。结果bFGF 可明显抑制人椎间盘细胞外基质成分Aggrecan 和Ⅱ型胶原mRNA的表达(P<0.05)。Real-time RT-PCR和Western blot 方法均提示bFGF 可抑制ChM-1 的表达水平(P<0.05),并具有剂量依赖性。结论bFGF 在发挥分解代谢功能的同时还具有潜在的促进血管生长作用。

关键词: 椎间盘, 碱性成纤维细胞生长因子, 软骨调节素, 退变,

Abstract: Objective To investigate the effect of basic fibroblast growth factor (bFGF) on the synthesis of extracellular matrixc (ECM)and expression of chondromodulin in human intervertebral disc cells. Methods 4 intervertebral discs (IVDs) obtained from patients in thetreatment of disc degenerative disease were used for cell culture. The secondary generation of intervertebral disc cells were cultured for 7days, then different concentration of bFGF (0, 0.1 ng/ml, 1 ng/ml, 10 ng/ml)were added to the medium and treated for 72 hours. Real-timeRT-PCR was used to detect the change of Aggrecan and type Ⅱ collagen mRNA expression. The effect of FGF on the expression of ChM-1,a cartilage derived anti-angiogenic factor, was also used by means of Real-time RT-PCR and Western blot. Results Real-time RT-PCRshowed that bFGF can significantly inhibit the expression of Aggrecan and type Ⅱ collagen mRNA. Both Real-time RT-PCR and Westernblot showed that the expression of ChM-1 was down-regulated by administration of bFGF with dose-dependent way. Conclusion bFGFserves primarily as a catabolic factor and induce the angiogenesis in the process of intervertebral disc degeneration.

Key words: intervertebral disc, basic fibroblast growth factor, chondromodulin, degeneration, human