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.
Similar content being viewed by others
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
References
Akar E, Somay H (2019) Comparative morphometric analysis of congenital and acquired lumbar spinal stenosis. J Clin Neurosci 68:256–261
AlSudais H, Wiper-Bergeron N (2019) Contaminating reactivity of a monoclonal CCAAT/Enhancer Binding Protein beta antibody in differentiating myoblasts. BMC Res Notes 12:717
Buck M, Chojkier M (2007) C/EBPbeta associates with caspase 8 complex proteins and modulates apoptosis in hepatic stellate cells. J Clin Gastroenterol 41(Suppl 3):S295–299
Catheline SE, Hoak D, Chang M, Ketz JP, Hilton MJ, Zuscik MJ, Jonason JH (2019) Chondrocyte-specific RUNX2 overexpression accelerates post-traumatic osteoarthritis progression in adult mice. J Bone Miner Res 34:1676–1689
Chanalaris A, Clarke H, Guimond SE, Vincent TL, Turnbull JE, Troeberg L (2019) Heparan sulfate proteoglycan synthesis is dysregulated in human osteoarthritic cartilage. Am J Pathol 189:632–647
Chen C, Bao GF, Xu G, Sun Y, Cui ZM (2018) Altered Wnt and NF-kappaB signaling in facet joint osteoarthritis: insights from RNA deep sequencing. Tohoku J Exp Med 245:69–77
Chowdhury SK, Byrne RM, Zhou Y, Zhang X (2018) Lumbar facet joint kinematics and load effects during dynamic lifting. Hum Factors 60:1130–1145
Eubanks JD, Toy JO, Messerschmitt P, Cooperman DR, Ahn NU (2009) Anatomic variance of interfacet distance and its relationship to facet arthrosis and disk degeneration in the lumbar spine. Orthopedics 32:893
Faber F (2019) Lumbar facet joint disease: classification, clinical diagnostics, and minimally invasive treatment. Orthopade 48:77–83
Guo CH, Yue ZP, Bai ZK, Li DD, Yang ZQ, Guo B (2016) Runx2 acts downstream of C/EBPbeta to regulate the differentiation of uterine stromal cells in mice. Cell Tissue Res 366:393–401
Hayashida M, Okazaki K, Fukushi J, Sakamoto A, Iwamoto Y (2009) CCAAT/enhancer binding protein beta mediates expression of matrix metalloproteinase 13 in human articular chondrocytes in inflammatory arthritis. Arthritis Rheum 60:708–716
Hirata M, Kugimiya F, Fukai A, Saito T, Yano F, Ikeda T, Mabuchi A, Sapkota BR, Akune T, Nishida N, Yoshimura N, Nakagawa T, Tokunaga K, Nakamura K, Chung UI, Kawaguchi H (2012) C/EBPbeta and RUNX2 cooperate to degrade cartilage with MMP-13 as the target and HIF-2alpha as the inducer in chondrocytes. Hum Mol Genet 21:1111–1123
Huang E, Huang H, Guan T, Liu C, Qu D, Xu Y, Yang J, Yan L, Xiong Y, Liang T, Wang Q, Chen L (2019) Involvement of C/EBPbeta-related signaling pathway in methamphetamine-induced neuronal autophagy and apoptosis. Toxicol Lett 312:11–21
Hwang HS, Kim HA (2015) Chondrocyte apoptosis in the pathogenesis of osteoarthritis. Int J Mol Sci 16:26035–26054
Jiang J, Zhang J, Wu C, Guo X, Chen C, Bao G, Sun Y, Chen J, Xue P, Xu G, Cui Z (2018) Up-regulation of TRAF2 inhibits chondrocytes apoptosis in lumbar facet joint osteoarthritis. Biochem Biophys Res Commun 503:1659–1665
Ko S, Vaccaro AR, Lee S, Lee J, Chang H (2014) The prevalence of lumbar spine facet joint osteoarthritis and its association with low back pain in selected Korean populations. Clin Orthop Surg 6:385–391
Komori T (2018) Runx2, an inducer of osteoblast and chondrocyte differentiation. Histochem Cell Biol 149:313–323
Li D, Liu J, Huang S, Bi X, Wang B, Chen Q, Chen H, Pu X (2018) CCAAT enhancer binding protein beta promotes tumor growth and inhibits apoptosis in prostate cancer by methylating estrogen receptor beta. Neoplasma 65:34–41
Li J, Ding Z, Li Y, Wang W, Wang J, Yu H, Liu A, Miao J, Chen S, Wu T, Cao Y (2019) BMSCs-derived exosomes ameliorate pain via abrogation of aberrant nerve invasion in subchondral bone in lumbar facet joint osteoarthritis. J Orthop Res. 38:670–679
Li JJ, Johnson AR (2011) Selective MMP13 inhibitors. Med Res Rev 31:863–894
Linov L, Klindukhov A, Li L, Kalichman L (2013) Lumbar facet joint orientation and osteoarthritis: a cross-sectional study. J Back Musculoskelet Rehabil 26:421–426
Malemud CJ (2017) Negative regulators of JAK/STAT signaling in rheumatoid arthritis and osteoarthritis. Int J Mol Sci 18:482
Mann SJ, Singh P (2019) Lumbar facet arthropathy. Arthritis Rheum 59(11):1618–1624
Marchildon F, Fu D, Lala-Tabbert N, Wiper-Bergeron N (2016) CCAAT/enhancer binding protein beta protects muscle satellite cells from apoptosis after injury and in cancer cachexia. Cell Death Dis 7:e2109
Marshall J, Dolan BM, Garcia EP, Sathe S, Tang X, Mao Z, Blair LA (2003) Calcium channel and NMDA receptor activities differentially regulate nuclear C/EBPbeta levels to control neuronal survival. Neuron 39:625–639
Mehrbod P, Ande SR, Alizadeh J, Rahimizadeh S, Shariati A, Malek H, Hashemi M, Glover KKM, Sher AA, Coombs KM, Ghavami S (2019) The roles of apoptosis, autophagy and unfolded protein response in arbovirus, influenza virus, and HIV infections. Virulence 10:376–413
Morimoto M, Higashino K, Manabe H, Tezuka F, Yamashita K, Takata Y, Takao S, Sakai T, Chikawa T, Nagamachi A, Sairyo K (2019) Age-related changes in axial and sagittal orientation of the facet joints: Comparison with changes in degenerative spondylolisthesis. J Orthop Sci 24:50–56
Shu S, Xu Y, Xie L, Ouyang Y (2017) The role of C/EBPbeta phosphorylation in modulating membrane phospholipids repairing in LPS-induced human lung/bronchial epithelial cells. Gene 629:76–85
Ushijima T, Okazaki K, Tsushima H, Iwamoto Y (2014) CCAAT/enhancer-binding protein beta regulates the repression of type II collagen expression during the differentiation from proliferative to hypertrophic chondrocytes. J Biol Chem 289:2852–2863
Wang M, Sampson ER, Jin H, Li J, Ke QH, Im HJ, Chen D (2013) MMP13 is a critical target gene during the progression of osteoarthritis. Arthritis Res Ther 15:R5
Wang Y, Xiang C, Sun X, Wu S, Lv J, Li P, Wei X, Wei L (2019) DAla2GIP antagonizes H2O2-induced chondrocyte apoptosis and inflammatory factor secretion. Bone 127:656–663
Zahid MDK, Rogowski M, Ponce C, Choudhury M, Moustaid-Moussa N, Rahman SM (2019) CCAAT/enhancer-binding protein beta (C/EBPbeta) knockdown reduces inflammation, ER stress, and apoptosis, and promotes autophagy in oxLDL-treated RAW2647 macrophage cells. Mol Cell Biochem 463:211–223
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).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflicts of interest
The authors declare no conflicts of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10735-020-09891-8