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Interaction between C/EBPβ and RUNX2 promotes apoptosis of chondrocytes during human lumbar facet joint degeneration

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Abstract

The pathophysiological changes in cartilage are a crucial feature of lumbar facet joint (LFJ) degeneration and arthritis. However, the molecular mechanism of human LFJ degeneration remains largely defined. This study aimed to examine the changes in chondrocytes at different stages of degenerative LFJ using hematoxylin and eosin and Safranin O staining. The significant loss of chondrocytes in grades 2 and 3 of LFJs was observed. The expression levels of CCAAT enhancer binding protein β (C/EBPβ), Runt-related transcription factor 2 (RUNX2), and matrix metalloproteinase 13 (MMP13) also increased with the aggravation of degeneration (4.89, 5.77, and 6.3 times by Western blot). In vitro, chondrocytes scraped from the LFJs during surgery were stimulated by interleukin (IL)-1β to establish the injury model. The association of C/EBPβ and RUNX2 with active caspase-3 on chondrocytes was analyzed. The high expression level of C/EBPβ, RUNX2, and MMP13 was consistent with that of caspase-3, which reached a peak after 36 h of stimulation. Immunofluorescence suggested that C/EBPβ, RUNX2, and MMP13 co-labeled with active caspase-3. Moreover, immunoprecipitation data prompted that C/EBPβ was able to interact with RUNX2. The knockdown of C/EBPβ significantly decreased the expression levels of MMP13 and active caspase-3 (2.48 and 2.89 times as detected by Western blot analysis) and inhibited chondrocyte apoptosis, which was further demonstrated using flow cytometry. Taken together, the findings of this study uncovered that C/EBPβ could interact with RUNX2 to induce chondrocyte apoptosis in human LFJ degeneration by regulating the expression of MMP13.

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Abbreviations

OARSI:

Osteoarthritis research society international

LFJ:

Lumbar facet joint

PAGE:

Polyacrylamide gel electrophoresis

CT:

Computed tomography

BSA:

Bovine serum albumin

DAB:

Diaminobenzidin

C/EBPβ:

CCAAT/enhancer binding protein beta

RUNX2:

Runt-related transcription factor 2

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Acknowledgements

This work was supported by Jiangsu Province Young Medical Key Talents Project of China (QNRC2016407), Nantong "226 High-level Talents" Project (2018 III-341), Nantong City Science and Technology Project (MS22016007, MSZ18090, JC2018020), Clinical Medicine Project of Nantong University (2019JQ004), Programs of Jiangsu “Six Talents Peak” High-level Talent Project (WSW-277).

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  1. Jinlong Zhang and Jiawei Jiang have contributed equally to the work.

Authors and Affiliations

  1. Department of Spine Surgery, Nantong City No.1 People’s Hospital and Second Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, China

    Jinlong Zhang, Jiawei Jiang, Guofeng Bao, Guanhua Xu, Lingling Wang, Jiajia Chen, Chu Chen, Chunshuai Wu, Pengfei Xue, Dawei Xu, Yuyu Sun & Zhiming Cui

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  1. Jinlong Zhang
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Correspondence to Yuyu Sun or Zhiming Cui.

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Zhang, J., Jiang, J., Bao, G. et al. Interaction between C/EBPβ and RUNX2 promotes apoptosis of chondrocytes during human lumbar facet joint degeneration. J Mol Hist 51, 401–410 (2020). https://doi.org/10.1007/s10735-020-09891-8

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