Cargando…

Elevated lymphotoxin-α (TNFβ) is associated with intervertebral disc degeneration

BACKGROUND: Intervertebral disc degeneration (IVDD) is a primary cause of degenerative disc diseases; however, the mechanisms underlying the degeneration remain unclear. The immunoinflammatory response plays an important role in IVDD progression. The inflammatory cytokine lymphotoxin-α (LTα), former...

Descripción completa

Detalles Bibliográficos
Autores principales: Guo, Zhu, Qiu, Chensheng, Mecca, Christina, Zhang, Yang, Bian, Jiang, Wang, Yan, Wu, Xiaolin, Wang, Tianrui, Su, Weiliang, Li, Xianglin, Zhang, Wei, Chen, Bohua, Xiang, Hongfei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7807514/
https://www.ncbi.nlm.nih.gov/pubmed/33441130
http://dx.doi.org/10.1186/s12891-020-03934-7
_version_ 1783636757204434944
author Guo, Zhu
Qiu, Chensheng
Mecca, Christina
Zhang, Yang
Bian, Jiang
Wang, Yan
Wu, Xiaolin
Wang, Tianrui
Su, Weiliang
Li, Xianglin
Zhang, Wei
Chen, Bohua
Xiang, Hongfei
author_facet Guo, Zhu
Qiu, Chensheng
Mecca, Christina
Zhang, Yang
Bian, Jiang
Wang, Yan
Wu, Xiaolin
Wang, Tianrui
Su, Weiliang
Li, Xianglin
Zhang, Wei
Chen, Bohua
Xiang, Hongfei
author_sort Guo, Zhu
collection PubMed
description BACKGROUND: Intervertebral disc degeneration (IVDD) is a primary cause of degenerative disc diseases; however, the mechanisms underlying the degeneration remain unclear. The immunoinflammatory response plays an important role in IVDD progression. The inflammatory cytokine lymphotoxin-α (LTα), formerly known as TNFβ, is associated with various pathological conditions, while its role in the pathogenesis of IVDD remains elusive. METHODS: Real-time quantitative polymerase chain reaction (RT-qPCR), Western blotting (WB), and enzyme-linked immunosorbent assays were used to assess the levels of LTα in human nucleus pulposus (NP) tissues between degeneration and control groups. The plasma concentrations of LTα and C-reactive protein (CRP) were compared between healthy and IVDD patients. Rat primary NP cells were cultured and identified via immunofluorescence. Methyl-thiazolyl-tetrazolium assays and flow cytometry were used to evaluate the effects of LTα on rat NP cell viability. After NP cells were treated with LTα, degeneration-related molecules (Caspase-3, Caspase-1, matrix metalloproteinase (MMP) -3, aggrecan and type II collagen) were measured via RT-qPCR and WB. RESULTS: The levels of both the mRNA and protein of LTα in human degenerated NP tissue significantly increased. Plasma LTα and CRP did not differ between healthy controls and IVDD patients. Rat primary NP cells were cultured, and the purity of primary NP cells was > 90%. Cell experiments showed inversely proportional relationships among the LTα dose, treatment time, and cell viability. The optimal conditions (dose and time) for LTα treatment to induce rat NP cell degeneration were 5 μg/ml and 48 ~ 72 h. The apoptosis rate and the levels of Caspase-3, Caspase-1, and MMP-3 significantly increased after LTα treatment, while the levels of type II collagen and aggrecan were decreased, and the protein expression levels were consistent with their mRNA expression levels. CONCLUSIONS: This study demonstrated that elevated LTα is closely associated with IVDD and that LTα may induce NP cell apoptosis and reduce important extracellular matrix (ECM) proteins, which cause adverse effects on IVDD progress. Moreover, the optimal conditions for LTα treatment to induce NP cell degeneration were determined. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12891-020-03934-7.
format Online
Article
Text
id pubmed-7807514
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-78075142021-01-14 Elevated lymphotoxin-α (TNFβ) is associated with intervertebral disc degeneration Guo, Zhu Qiu, Chensheng Mecca, Christina Zhang, Yang Bian, Jiang Wang, Yan Wu, Xiaolin Wang, Tianrui Su, Weiliang Li, Xianglin Zhang, Wei Chen, Bohua Xiang, Hongfei BMC Musculoskelet Disord Research Article BACKGROUND: Intervertebral disc degeneration (IVDD) is a primary cause of degenerative disc diseases; however, the mechanisms underlying the degeneration remain unclear. The immunoinflammatory response plays an important role in IVDD progression. The inflammatory cytokine lymphotoxin-α (LTα), formerly known as TNFβ, is associated with various pathological conditions, while its role in the pathogenesis of IVDD remains elusive. METHODS: Real-time quantitative polymerase chain reaction (RT-qPCR), Western blotting (WB), and enzyme-linked immunosorbent assays were used to assess the levels of LTα in human nucleus pulposus (NP) tissues between degeneration and control groups. The plasma concentrations of LTα and C-reactive protein (CRP) were compared between healthy and IVDD patients. Rat primary NP cells were cultured and identified via immunofluorescence. Methyl-thiazolyl-tetrazolium assays and flow cytometry were used to evaluate the effects of LTα on rat NP cell viability. After NP cells were treated with LTα, degeneration-related molecules (Caspase-3, Caspase-1, matrix metalloproteinase (MMP) -3, aggrecan and type II collagen) were measured via RT-qPCR and WB. RESULTS: The levels of both the mRNA and protein of LTα in human degenerated NP tissue significantly increased. Plasma LTα and CRP did not differ between healthy controls and IVDD patients. Rat primary NP cells were cultured, and the purity of primary NP cells was > 90%. Cell experiments showed inversely proportional relationships among the LTα dose, treatment time, and cell viability. The optimal conditions (dose and time) for LTα treatment to induce rat NP cell degeneration were 5 μg/ml and 48 ~ 72 h. The apoptosis rate and the levels of Caspase-3, Caspase-1, and MMP-3 significantly increased after LTα treatment, while the levels of type II collagen and aggrecan were decreased, and the protein expression levels were consistent with their mRNA expression levels. CONCLUSIONS: This study demonstrated that elevated LTα is closely associated with IVDD and that LTα may induce NP cell apoptosis and reduce important extracellular matrix (ECM) proteins, which cause adverse effects on IVDD progress. Moreover, the optimal conditions for LTα treatment to induce NP cell degeneration were determined. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12891-020-03934-7. BioMed Central 2021-01-13 /pmc/articles/PMC7807514/ /pubmed/33441130 http://dx.doi.org/10.1186/s12891-020-03934-7 Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Guo, Zhu
Qiu, Chensheng
Mecca, Christina
Zhang, Yang
Bian, Jiang
Wang, Yan
Wu, Xiaolin
Wang, Tianrui
Su, Weiliang
Li, Xianglin
Zhang, Wei
Chen, Bohua
Xiang, Hongfei
Elevated lymphotoxin-α (TNFβ) is associated with intervertebral disc degeneration
title Elevated lymphotoxin-α (TNFβ) is associated with intervertebral disc degeneration
title_full Elevated lymphotoxin-α (TNFβ) is associated with intervertebral disc degeneration
title_fullStr Elevated lymphotoxin-α (TNFβ) is associated with intervertebral disc degeneration
title_full_unstemmed Elevated lymphotoxin-α (TNFβ) is associated with intervertebral disc degeneration
title_short Elevated lymphotoxin-α (TNFβ) is associated with intervertebral disc degeneration
title_sort elevated lymphotoxin-α (tnfβ) is associated with intervertebral disc degeneration
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7807514/
https://www.ncbi.nlm.nih.gov/pubmed/33441130
http://dx.doi.org/10.1186/s12891-020-03934-7
work_keys_str_mv AT guozhu elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT qiuchensheng elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT meccachristina elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT zhangyang elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT bianjiang elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT wangyan elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT wuxiaolin elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT wangtianrui elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT suweiliang elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT lixianglin elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT zhangwei elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT chenbohua elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration
AT xianghongfei elevatedlymphotoxinatnfbisassociatedwithintervertebraldiscdegeneration