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周期性牵张应力对人椎间盘纤维环细胞合成和分解代谢的研究

范东伟1,陈仲强2,郭昭庆3,齐强3,李危石3   

  1. 1. 北京大学第三医院生殖中心地下一层B15房间（于洋 收）
    2. 北医三院
    3. 北京大学第三医院骨科（北京市海淀区花园北路49号）
  • 收稿日期:2013-12-24 修回日期:2014-01-08 出版日期:2014-03-15 发布日期:2014-03-15
  • 通讯作者: 陈仲强

Effects of different magnitudes of mechanical stretch on human intervertebral disc cells

  • Received:2013-12-24 Revised:2014-01-08 Published:2014-03-15 Online:2014-03-15

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摘要： 目的：探讨不同牵张应力对体外培养的人椎间盘纤维环细胞合成和分解代谢的研究。 方法：采用Flexercell Strain Unit对人椎间盘纤维环细胞施加不同拉伸幅度的周期性牵张应力，通过real-time PCR，Western Blot，免疫组化以及阻断剂等方法，检测细胞合成代谢基因(collagen-1A1，collagen-2A1，aggrecan，versican)和分解代谢基因(MMP-3，MMP-13，ADAMTS-4，ADAMTS-5)在周期性牵张应力下椎间盘纤维环细胞中的表达。 结果：合成代谢基因中collagen-1A1，collagen-2A1和aggrecan，但不是versican，在12%的应力条件下表达升高，在18%表达下降，相反分解代谢基因MMP-3，MMP-13，ADAMTS-4和ADAMTS-5在18%的应变下表达升高，进一步研究发现ADAMTS-4表达随应力拉伸幅度升高而表达增强。进一步研究发现ERK1/2的阻断剂U0126，能明显阻断应力诱导的ADAMTS-4的表达，而p38和JNK的阻断剂SP6000125和SB203580阻断应力诱导ADAMTS-4表达作用不明显。 结论：不同拉伸幅度的牵张应力对椎间盘纤维化细胞的生物学行为的影响不同，其结果可以影响椎间盘的退变。

关键词: 应力, 椎间盘细胞, ADAMTS-4, ERK

Abstract: ObjectiveMechanical stress plays an important role in intervertebral disc degeneration which is closely related with low back pain. The effects of mechanical stress on disc degeneration and the mechanisms involved are poorly understood. In the present study, we investigated how different magnitudes of mechanical stretch affected human intervertebral disc cells. MethodsCells were subjected to 0%, 6%, 12%, or 18% elongation for 24 h using a Flexercell Strain Unit, and the mRNA expressions of anabolic genes (collagen-1A1, collagen-2A1, aggrecan, versican) and catabolic genes (MMP-3, MMP-13, ADAMTS-4, ADAMTS-5) were examined. ResultsThe results showed that the expression of collagen-1A1, collagen-2A1 and aggrecan, but not versican, were increased at 12%, and decreased at 18%, whereas catabolic genes: MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5, were increased at 18%. Interestingly, mechanical stretch induced a magnitude-dependent increase in ADAMTS-4 synthesis. Furthermore, the effect seems to be finely tuned by stretching-triggered activation of distinct mitogen-activated protein kinase cascades. Specifically, an ERK1/2specific inhibitor, U0126, significantly inhibited stretching-induced ADAMTS-4 expression, whereas the inhibitors of p38 and JNK, SP6000125 and SB203580 only slightly decreased stretching-induced ADAMTS-4 expression. ConclusionThese results indicate that different magnitudes of mechanical stretch influence the biological behavior of intervertebral disc cells, which profoundly affects intervertebral disc degeneration.

Key words: Mechanical stress, Intervertebral disc cells, ADAMTS-4, ERK