Expression of AMP-activated protein kinase in subcultured rat endplate chondrocytes

doi:10.3969/j.issn.2095-4344.2016.29.006

Abstract

Abstract:

BACKGROUND: Endplate cartilage degeneration initiates intervertebral disc degeneration. AMP-activated protein kinase (AMPK) regulates the formation and degradation of cartilage.
OBJECTIVE: To explore the role of AMPK in an in vitro natural degeneration model of chondrocytes derived from endplate of rat intervertebral discs.
METHODS: Morphology of in vitro subcultured endplate chondrocytes of rat intervertebral discs at passages 0, 2, and 5 were observed under an inverted microscope following cytoskeleton staining. Chondrocyte phenotype, proliferation, and the cartilage marker genes (type II collagen, proteoglycan, SOX-9, matrix metalloproteinase-3 and -13), and AMPK phosphorylation were determined by toluidine blue staining, MTT assay, real-time PCR analysis, and western blot assay, respectively.
RESULTS AND CONCLUSION: The altered morphology, decreased proliferation ability, and phenotype loss were observed in chondrocytes with increased passage number. Gene expression of type II collagen, proteoglycan, SOX-9 was significantly decreased; while gene expression of matrix metalloproteinase-3 and -13 was significantly increased in endplate chondrocytes at passage 5 compared with those at passages 0 and 2. AMPK phosphorylation in endplate chondrocytes at passage 5 was significantly decreased. These findings indicate that AMPK phosphorylation is involved in in vitro natural degeneration of chondrocytes derived from the endplate of rat intervertebral discs, and the degeneration of endplate chondrocytes and intervertebral discs can be inhibited through the regulation of AMPK activity.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Intervertebral Disk, Chondrocytes, Matrix Metalloproteinases

CLC Number:

R318

Zhao Quan-lai, Zheng Quan, Xu Hong-guang, Shen Xiang, Wang Hong, Liu Ping, Wang Ling-ting, . Expression of AMP-activated protein kinase in subcultured rat endplate chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(29): 4297-4302.

Figures/Tables 3

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