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Attenuation of the degenerative effects of endothelin-1 on cartilaginous end plate cells by the endothelin receptor antagonist BQ-123 via the Wnt/β-catenin signaling pathway.
Spine J 2018; 18(9):1669-1677SJ

Abstract

BACKGROUND CONTEXT

Endothelin-1 (ET-1) is an inflammatory mediator associated with cartilage end plate (CEP) degeneration in the intervertebral disc (IVD). SOX9 is downregulated during CEP degeneration, along with its targets, collagen II and aggrecan. Wnt/β-catenin signaling is associated with CEP degeneration and a downstream target of SOX9; however, the precise mechanism of CEP degeneration and the role of ET-1 are largely unknown.

PURPOSE

The purpose of the study was to evaluate the influence of the endothelin-A receptor antagonist, BQ-123, on ET-1-induced effects on cartilaginous end plate cells (CECs) associated with CEP degeneration via the Wnt/β-catenin signaling pathway.

STUDY DESIGN/SETTING

The influence of ET-1 on the expression levels of collagen II, aggrecan, and SOX9 in CECs and the effect of BQ-123 in this context were investigated.

METHODS

To establish a model for CEP degeneration, three lumbar discs (L3-L4, L4-L5, and L5-L6 levels) in New Zealand white rabbits were punctured close to the vertebral end plate using a 14G needle. Intervertebral disc degeneration was evaluated by magnetic resonance imaging 4 weeks after vertebral end plate injury. CECs were then isolated from the degenerated CEPs to allow evaluation of the role of ET-1 and BQ-123 and to investigate their effects on the Wnt/β-catenin signaling pathway. The expression of ET-1 in CECs from degenerated CEPs was analyzed by immunofluorescent staining. Changes in the levels of collagen II, aggrecan, and SOX9 were evaluated in CECs by real-time polymerase chain reaction and by Western blotting. The Wnt/β-catenin signaling pathway was also investigated by Western blotting.

RESULTS

After 4 weeks, IVDs with vertebral end plate injury exhibited clear signs of disc degeneration. Immunofluorescent staining showed that ET-1 was expressed in the cytoplasm of CECs. Endothelin-1 stimulation significantly inhibited the expression of collagen II, aggrecan, and SOX9 in CECs, whereas BQ-123 increased the levels of these three molecules. In addition, ET-1 stimulation increased the expression of β-catenin, cyclin D1, and Dvl1 in the Wnt/β-catenin signaling pathway of CECs from degenerated discs and reduced the expression of GSK-3β, whereas BQ-123 had the opposite effect.

CONCLUSIONS

Endothelin-1 can reduce levels of collagen II, aggrecan, and SOX9 in CECs through activation of the Wnt/β-catenin signaling pathway, whereas BQ-123 attenuates these negative effects, highlighting a new molecular mechanism with potential for exploitation for treatment of CEP degeneration.

Authors+Show Affiliations

Department of Orthopaedic Oncology, Spinal Tumor Center, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China.Department of Orthopaedic Oncology, Spinal Tumor Center, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China.Department of Orthopaedic Oncology, Spinal Tumor Center, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China.Shanghai University of Medicine and Health Sciences Affiliated Zhoupu Hospital, Shanghai, 201318, China.Department of Orthopaedic Oncology, Spinal Tumor Center, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China.Department of Orthopaedic Oncology, Spinal Tumor Center, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China.Key Laboratory of Adolescent Health Assessment and Exercise Intervention Ministry of Education, East China Normal University, Shanghai, 200241, China.Department of Orthopaedic Oncology, Spinal Tumor Center, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China. Electronic address: Jianruxiao83@163.com.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

29886166

Citation

Yuan, Wei, et al. "Attenuation of the Degenerative Effects of Endothelin-1 On Cartilaginous End Plate Cells By the Endothelin Receptor Antagonist BQ-123 Via the Wnt/β-catenin Signaling Pathway." The Spine Journal : Official Journal of the North American Spine Society, vol. 18, no. 9, 2018, pp. 1669-1677.
Yuan W, Li ZX, Zhao CL, et al. Attenuation of the degenerative effects of endothelin-1 on cartilaginous end plate cells by the endothelin receptor antagonist BQ-123 via the Wnt/β-catenin signaling pathway. Spine J. 2018;18(9):1669-1677.
Yuan, W., Li, Z. X., Zhao, C. L., Hou, T. H., Hu, S. W., Liu, W. B., ... Xiao, J. R. (2018). Attenuation of the degenerative effects of endothelin-1 on cartilaginous end plate cells by the endothelin receptor antagonist BQ-123 via the Wnt/β-catenin signaling pathway. The Spine Journal : Official Journal of the North American Spine Society, 18(9), pp. 1669-1677. doi:10.1016/j.spinee.2018.05.012.
Yuan W, et al. Attenuation of the Degenerative Effects of Endothelin-1 On Cartilaginous End Plate Cells By the Endothelin Receptor Antagonist BQ-123 Via the Wnt/β-catenin Signaling Pathway. Spine J. 2018;18(9):1669-1677. PubMed PMID: 29886166.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Attenuation of the degenerative effects of endothelin-1 on cartilaginous end plate cells by the endothelin receptor antagonist BQ-123 via the Wnt/β-catenin signaling pathway. AU - Yuan,Wei, AU - Li,Zhen-Xi, AU - Zhao,Cheng-Long, AU - Hou,Tian-Hui, AU - Hu,Si-Wang, AU - Liu,Wei-Bo, AU - Yuan,Feng-Lai, AU - Xiao,Jian-Ru, Y1 - 2018/06/07/ PY - 2018/01/17/received PY - 2018/04/06/revised PY - 2018/05/03/accepted PY - 2018/6/11/pubmed PY - 2019/3/26/medline PY - 2018/6/11/entrez KW - BQ-123 KW - Cartilaginous end plate cells KW - Degeneration KW - Endothelin-1 KW - Intervertebral disc KW - Wnt/β-catenin signaling pathway SP - 1669 EP - 1677 JF - The spine journal : official journal of the North American Spine Society JO - Spine J VL - 18 IS - 9 N2 - BACKGROUND CONTEXT: Endothelin-1 (ET-1) is an inflammatory mediator associated with cartilage end plate (CEP) degeneration in the intervertebral disc (IVD). SOX9 is downregulated during CEP degeneration, along with its targets, collagen II and aggrecan. Wnt/β-catenin signaling is associated with CEP degeneration and a downstream target of SOX9; however, the precise mechanism of CEP degeneration and the role of ET-1 are largely unknown. PURPOSE: The purpose of the study was to evaluate the influence of the endothelin-A receptor antagonist, BQ-123, on ET-1-induced effects on cartilaginous end plate cells (CECs) associated with CEP degeneration via the Wnt/β-catenin signaling pathway. STUDY DESIGN/SETTING: The influence of ET-1 on the expression levels of collagen II, aggrecan, and SOX9 in CECs and the effect of BQ-123 in this context were investigated. METHODS: To establish a model for CEP degeneration, three lumbar discs (L3-L4, L4-L5, and L5-L6 levels) in New Zealand white rabbits were punctured close to the vertebral end plate using a 14G needle. Intervertebral disc degeneration was evaluated by magnetic resonance imaging 4 weeks after vertebral end plate injury. CECs were then isolated from the degenerated CEPs to allow evaluation of the role of ET-1 and BQ-123 and to investigate their effects on the Wnt/β-catenin signaling pathway. The expression of ET-1 in CECs from degenerated CEPs was analyzed by immunofluorescent staining. Changes in the levels of collagen II, aggrecan, and SOX9 were evaluated in CECs by real-time polymerase chain reaction and by Western blotting. The Wnt/β-catenin signaling pathway was also investigated by Western blotting. RESULTS: After 4 weeks, IVDs with vertebral end plate injury exhibited clear signs of disc degeneration. Immunofluorescent staining showed that ET-1 was expressed in the cytoplasm of CECs. Endothelin-1 stimulation significantly inhibited the expression of collagen II, aggrecan, and SOX9 in CECs, whereas BQ-123 increased the levels of these three molecules. In addition, ET-1 stimulation increased the expression of β-catenin, cyclin D1, and Dvl1 in the Wnt/β-catenin signaling pathway of CECs from degenerated discs and reduced the expression of GSK-3β, whereas BQ-123 had the opposite effect. CONCLUSIONS: Endothelin-1 can reduce levels of collagen II, aggrecan, and SOX9 in CECs through activation of the Wnt/β-catenin signaling pathway, whereas BQ-123 attenuates these negative effects, highlighting a new molecular mechanism with potential for exploitation for treatment of CEP degeneration. SN - 1878-1632 UR - https://www.unboundmedicine.com/medline/citation/29886166/Attenuation_of_the_degenerative_effects_of_endothelin_1_on_cartilaginous_end_plate_cells_by_the_endothelin_receptor_antagonist_BQ_123_via_the_Wnt/β_catenin_signaling_pathway_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1529-9430(18)30217-1 DB - PRIME DP - Unbound Medicine ER -