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Association of endothelin-1 expression and cartilaginous endplate degeneration in humans.
PLoS One 2013; 8(4):e60062Plos

Abstract

BACKGROUND

Inflammatory cytokines are involved in intervertebral disc (IVD) degeneration. Endothelin-1 (ET-1), a 21-amino-acid cytokine implicated with cartilage degradation, is secreted by vascular endothelial cells and also by many other cell types. The expression of ET-1 in human IVD cartilage endplate (CEP) and its role in disc degeneration have not been explored.

METHODS AND FINDINGS

The expression of ET-1 in degenerated CEP was analyzed by immunohistochemical staining and Western blotting; ET-1 was demonstrated in cartilaginous endplate cells (CECs) by immunofluorescent staining. The ET-1 mRNA expression and protein production by CECs stimulated by tumor necrosis factor alpha (TNF-α), a pro-inflammatory cytokine, were determined by real-time PCR analysis and Western blotting, respectively. The matrix metalloprotease-1 (MMP-1), MMP-13 and tissue inhibitor of metalloproteases-1 (TIMP-1) levels in the supernatant of cultured CECs treated with ET-1 were determined using enzyme-linked immunosorbent assays. Nitric oxide (NO) release and nitric oxide synthase (NOS) activity were measured using a spectrophotometric assay. The apoptosis of CECs by ET-1 was measured by an Annexin V-FITC detection assay. The production of ET-1 in degenerated cartilage endplate was significantly higher than normal CEP. The results showed that ET-1 was expressed by CECs and modulated by TNF-α in a dose-dependent manner. ET-1 increased production of MMP-1 and MMP-13, decreased TIMP-1 production, and induced NO and NOS release by cultured CECs. The direct stimulation of CECs by ET-1 did not promote cell apoptosis.

CONCLUSION

The study results suggest that ET-1 played a pivotal role in human CEP degeneration, and may be a new target for development of therapies for this condition.

Authors+Show Affiliations

Department of Orthopedic Surgery, Zhongshan Hospital, Fudan University, Shanghai, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

23565184

Citation

Yuan, Wei, et al. "Association of Endothelin-1 Expression and Cartilaginous Endplate Degeneration in Humans." PloS One, vol. 8, no. 4, 2013, pp. e60062.
Yuan W, Zhao MD, Yuan FL, et al. Association of endothelin-1 expression and cartilaginous endplate degeneration in humans. PLoS ONE. 2013;8(4):e60062.
Yuan, W., Zhao, M. D., Yuan, F. L., Che, W., Duan, P. G., Liu, Y., & Dong, J. (2013). Association of endothelin-1 expression and cartilaginous endplate degeneration in humans. PloS One, 8(4), pp. e60062. doi:10.1371/journal.pone.0060062.
Yuan W, et al. Association of Endothelin-1 Expression and Cartilaginous Endplate Degeneration in Humans. PLoS ONE. 2013;8(4):e60062. PubMed PMID: 23565184.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Association of endothelin-1 expression and cartilaginous endplate degeneration in humans. AU - Yuan,Wei, AU - Zhao,Ming-Dong, AU - Yuan,Feng-Lai, AU - Che,Wu, AU - Duan,Ping-Guo, AU - Liu,Yi, AU - Dong,Jian, Y1 - 2013/04/02/ PY - 2013/01/20/received PY - 2013/02/21/accepted PY - 2013/4/9/entrez PY - 2013/4/9/pubmed PY - 2013/10/31/medline SP - e60062 EP - e60062 JF - PloS one JO - PLoS ONE VL - 8 IS - 4 N2 - BACKGROUND: Inflammatory cytokines are involved in intervertebral disc (IVD) degeneration. Endothelin-1 (ET-1), a 21-amino-acid cytokine implicated with cartilage degradation, is secreted by vascular endothelial cells and also by many other cell types. The expression of ET-1 in human IVD cartilage endplate (CEP) and its role in disc degeneration have not been explored. METHODS AND FINDINGS: The expression of ET-1 in degenerated CEP was analyzed by immunohistochemical staining and Western blotting; ET-1 was demonstrated in cartilaginous endplate cells (CECs) by immunofluorescent staining. The ET-1 mRNA expression and protein production by CECs stimulated by tumor necrosis factor alpha (TNF-α), a pro-inflammatory cytokine, were determined by real-time PCR analysis and Western blotting, respectively. The matrix metalloprotease-1 (MMP-1), MMP-13 and tissue inhibitor of metalloproteases-1 (TIMP-1) levels in the supernatant of cultured CECs treated with ET-1 were determined using enzyme-linked immunosorbent assays. Nitric oxide (NO) release and nitric oxide synthase (NOS) activity were measured using a spectrophotometric assay. The apoptosis of CECs by ET-1 was measured by an Annexin V-FITC detection assay. The production of ET-1 in degenerated cartilage endplate was significantly higher than normal CEP. The results showed that ET-1 was expressed by CECs and modulated by TNF-α in a dose-dependent manner. ET-1 increased production of MMP-1 and MMP-13, decreased TIMP-1 production, and induced NO and NOS release by cultured CECs. The direct stimulation of CECs by ET-1 did not promote cell apoptosis. CONCLUSION: The study results suggest that ET-1 played a pivotal role in human CEP degeneration, and may be a new target for development of therapies for this condition. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/23565184/Association_of_endothelin_1_expression_and_cartilaginous_endplate_degeneration_in_humans_ L2 - http://dx.plos.org/10.1371/journal.pone.0060062 DB - PRIME DP - Unbound Medicine ER -