Cyclic tensile stress of human annulus fibrosus cells induces MAPK activation: involvement in proinflammatory gene expression

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• 作者：H. Pratsinis^&dagger ; ; A. Papadopoulou^&dagger ; ; C. Neidlinger-Wilke^&Dagger ; ; M. Brayda-Bruno^§ ; ; H.-J. Wilke^&Dagger ; ; D. Kletsas^&dagger ; ; ^{dkletsas@bio.demokritos.gr" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Cyclic tensile stress ; Annulus fibrosus ; Mitogen-activated protein kinase ; Cyclooxygenase-2 ; Interleukin-6 ; Interleukin-8
• 刊名：Osteoarthritis and Cartilage
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：24
• 期：4
• 页码：679-687
• 全文大小：1005 K

文摘

To study the role of mitogen-activated protein kinases (MAPKs) in human annulus fibrosus (AF) cells subjected to cyclic tensile stress (CTS).

Design

An in vitro system for CTS studies was established using AF cultures on fibronectin-coated silicone dishes. MAPK phosphorylation was studied by western analysis, while gene expression was followed by qRT-PCR. DNA synthesis was assessed by both tritiated thymidine incorporation and flow cytometry, and collagen synthesis using tritiated proline incorporation and the protease-free collagenase method.

Results

All three MAPKs studied, i.e., ERK, SAPK/JNK, and p38 were found to be phosphorylated immediately after CTS application within physiological range. A second wave of phosphorylation appeared at later time points. MAPK activation was elevated at higher CTS magnitudes, but independent of the frequency. CTS did not stimulate DNA synthesis neither extracellular matrix turnover, but it stimulated the proinflammatory genes, COX-2, IL-6, and IL-8. This stimulation was more intense at the highest magnitude (8%) tested and at the median frequency (1 Hz) and time interval (12 h). Blocking of ERK, SAPK/JNK, and p38 MAPK inhibited the CTS-induced stimulation of COX-2 and IL-8, while IL-6 expression was mediated only by SAPK/JNK and p38 MAPK.

Conclusions

We have described for the first time the activation of MAPKs in human AF cells in response to CTS and showed that it drives an inflammatory reaction. These observations shed light on the mechanisms of intervertebral disc (IVD) cell responses to mechanical stress, contributing to the understanding of disc pathophysiology and possibly to the design of novel therapeutic interventions.