JNK phosphorylation promotes natural degeneration of cervical endplate chondrocytes by down-regulating expression of ANK

H.-G. Xu, J.-F. Cheng, H.-X. Peng, K. Lv, H. Wang, P. Liu, M. Zhong, M.-Y. Zhang

Department of Orthopedic Surgery, Yijishan Hospital, Wannan Medical College, Wuhu, People’s Republic of China. pumchxuhg@126.com

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• Orthopedics

BACKGROUND: Endplate degeneration leads to accelerated degeneration of the intervertebral disc. The importance of endplate chondrocytes in this process is unclear. Many cellular processes in chondrocytes are controlled by activated c-Jun N-terminal kinases (JNK) and protein kinase B (AKT). However, the involvement of their pathways in the degeneration process needs to be elucidated.

AIM: To study activation of JNK and AKT signaling pathways and their significance for degeneration of endplate chondrocytes, as well as involvement of progressive ankylosis protein (ANK) in this process.

MATERIALS AND METHODS: Rat primary chondrocytes were grown to confluence and subcultured until passage 4. Morphological appearances (microscope, hematoxylin & eosin staining, toluidine blue staining) and proliferation rates of cells (MTT test) were observed. Further, levels of type II collagen, aggrecan, phosphorylated JNK and AKT, total JNK, AKT and ANK were evaluated by qPCR, flow cytometry and Western blot assays. Furthermore, inhibition experiments with SP600125, the JNK inhibitor, were carried out in the passage 4 cells to assess the effects of the JNK pathway on natural degeneration of endplate chondrocytes.

RESULTS: The proliferative speed of endplate chondrocytes progressively decreased during passaging. Expressions of type II collagen and aggrecan were significantly decreased with cells at higher passages. Furthermore, phosphorylation of JNK, but not AKT, was significantly up-regulated and accompanied by reduced ANK expression. Inhibition of the JNK pathway increased expression of type II collagen, aggrecan and ANK and facilitated proliferation rates.

CONCLUSIONS: Phosphorylation of JNK promotes natural degeneration of cervical endplate chondrocytes, likely by down-regulating ANK expression.

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