Expression of NF-κB in a degenerative human intervertebral disc model

Abstract

Objective: To investigate the changes in the expression of NF-κB signaling pathway in human degenerative intervertebral discs. Methods: From October 2014 to March 2016, 55 nucleus pulposus of surgical patients with degenerative human intervertebral disc were collected for study in Department of Orthopedic Surgey, Hospital of Zaozhuang Mining Corporation, and Department of Orthopedics, Shanghai General Hospital Affiliated Shanghai Jiaotong University, School of Medicine.The collected nucleus pulposus tissues were divided into two groups: experimental group(30) and control group(25). Cell culture observed normal and degenerative nucleus pulposus cells morphological changes; immunofluorescence observed NF-κB p65 changes in the nucleus of nucleus pulposus cells.Real-time PCR was observed changes in aggregated proteoglycans and matrix metalloproteinase gene mRNA.Finally, the use of blockers of nucleus pulposus cells were treated 24 hours, Western blot analysis the changing of p65, ADAMTS-4, MMP-13, aggregate proteoglycans and collagen Ⅱ protein expression. Results: Compared with the experimental group, the nucleus pulposus cells in the control group had larger cell volume, abundant cytoplasm and faster growth rate.Cell immunofluorescence show Nondegenerative nucleus pulposus cells p65 protein was mainly localized in the cytoplasm, degeneration of nucleus pulposus cells p65 protein was mainly concentrated in the nucleus.RT-PCR showed degenerative group of matrix metalloproteinases (MMP-1, MMP-3, MMP-13), aggrecanase(ADAMTS-4, ADAMTS-5) and IL-6 mRNA expression was significantly higher than Nondegenerative group; aggrecan and type Ⅱ collagen expression than those without degeneration group was significantly lower.Expression of nucleus pulposus degeneration in nuclear protein p65 with the degenerative level increased gradually increased.BAY11-7082 blocked the activity of NF-κB signaling pathway, which could significantly down-regulate the expression of ADAMTS-4 and MMP-13 protein and significantly up-regulate the expression of Agg and COLⅡ protein.With the increase of BAY11-7082 concentration, gradually strengthened. Conclusion: The activation of the NF-κB signaling pathway in a degenerative intervertebral disc is gradually increased, regulating the over-expression of matrix-degrading enzymes.It plays an important role in the degradation of extra-cellular matrix.