open access

Vol 58, No 3 (2020)
Original paper
Submitted: 2020-06-03
Accepted: 2020-09-01
Published online: 2020-09-16
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RhTSG-6 inhibits IL-1β-induced extracellular matrix degradation and apoptosis by suppressing the p38, and JNK pathways in nucleus pulposus cells

Shishen Pei123, Jinwei Ying2, Yan Zhang2, Linhao Su2, Shi Cheng2, Dike Ruan12
DOI: 10.5603/FHC.a2020.0019
·
Pubmed: 32936927
·
Folia Histochem Cytobiol 2020;58(3):227-234.
Affiliations
  1. The Second School of Clinical Medicine, Southern Medical University, Guangzhou, China
  2. Department of Orthopedic, Navy General Hospital, Beijing, People’s Republic of China, China
  3. Department of Orthopedic Surgery, The 4th People’s Hospital of Hengshui City, Hebei, People’s Republic of China, China

open access

Vol 58, No 3 (2020)
ORIGINAL PAPERS
Submitted: 2020-06-03
Accepted: 2020-09-01
Published online: 2020-09-16

Abstract

Introduction. Intervertebral disc degeneration (IDD) is one of the major causes of low back pain (LBP) which seriously affects health and normal physical activity. Recombinant human tumor necrosis factor-a (TNF-a) induced protein 6 (rhTSG-6) has been reported to have therapeutic effects on a variety of inflammatory diseases, but the effect and mechanism of rhTSG-6 action in IDD are not fully understood. The present study was aimed to explore the functional role of rhTSG-6 in interleukin (IL)-1b-induced nucleus pulposus (NP) cell model.

Materials and methods. Experimental human NP cells were isolated from the patients with idiopathic scoliosis and treated with culture medium containing IL-1b (10 ng/mL) for 24 hours to induce extracellular matrix degradation and apoptosis, simulating an IDD model in vitro. The viability of NP cells was analyzed by the CCK-8 assay. The relevant mRNA and protein levels were measured by RT-qPCR and western blot. The apoptosis of NP cells was determined by flow cytometry analysis and western blot. Results. Compared with the NP cells without IL-1b treatment, IL-1b caused approximately 70% reduction in the viability of NP cells, while RhTSG-6 partly increased the decrease of IL-1b on cell viabilities. Moreover, treatment with rhTSG-6 considerably attenuated the upregulation of extracellular matrix (ECM)-catabolic factors (MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5), and increased the downregulation of ECM-anabolic factor (collagen II) in NP cells induced by IL-1b, indicating that ECM degradation was suppressed. Furthermore, rhTSG-6 also protected NP cells from IL-1b-induced apoptosis. Mechanically, rhTSG-6 inhibited the activation of members of mitogen-activated protein kinase (MAPK) pathway by blocking the phosphorylation of p38, c-Jun N-terminal kinase (JNK) and ERK in IL-1b-induced NP cells. Conclusions. RhTSG-6 can attenuate ECM degradation and apoptosis in IL-1b-induced NP cells by inhibiting the p38, JNK and ERK pathways, which may contribute to its potential application in the therapy of IDD.

Abstract

Introduction. Intervertebral disc degeneration (IDD) is one of the major causes of low back pain (LBP) which seriously affects health and normal physical activity. Recombinant human tumor necrosis factor-a (TNF-a) induced protein 6 (rhTSG-6) has been reported to have therapeutic effects on a variety of inflammatory diseases, but the effect and mechanism of rhTSG-6 action in IDD are not fully understood. The present study was aimed to explore the functional role of rhTSG-6 in interleukin (IL)-1b-induced nucleus pulposus (NP) cell model.

Materials and methods. Experimental human NP cells were isolated from the patients with idiopathic scoliosis and treated with culture medium containing IL-1b (10 ng/mL) for 24 hours to induce extracellular matrix degradation and apoptosis, simulating an IDD model in vitro. The viability of NP cells was analyzed by the CCK-8 assay. The relevant mRNA and protein levels were measured by RT-qPCR and western blot. The apoptosis of NP cells was determined by flow cytometry analysis and western blot. Results. Compared with the NP cells without IL-1b treatment, IL-1b caused approximately 70% reduction in the viability of NP cells, while RhTSG-6 partly increased the decrease of IL-1b on cell viabilities. Moreover, treatment with rhTSG-6 considerably attenuated the upregulation of extracellular matrix (ECM)-catabolic factors (MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5), and increased the downregulation of ECM-anabolic factor (collagen II) in NP cells induced by IL-1b, indicating that ECM degradation was suppressed. Furthermore, rhTSG-6 also protected NP cells from IL-1b-induced apoptosis. Mechanically, rhTSG-6 inhibited the activation of members of mitogen-activated protein kinase (MAPK) pathway by blocking the phosphorylation of p38, c-Jun N-terminal kinase (JNK) and ERK in IL-1b-induced NP cells. Conclusions. RhTSG-6 can attenuate ECM degradation and apoptosis in IL-1b-induced NP cells by inhibiting the p38, JNK and ERK pathways, which may contribute to its potential application in the therapy of IDD.

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Keywords

intervertebral disc degeneration; rhTSG-6; extracellular matrix degradation; apoptosis; IL-1β; nucleus pulposus cells

About this article
Title

RhTSG-6 inhibits IL-1β-induced extracellular matrix degradation and apoptosis by suppressing the p38, and JNK pathways in nucleus pulposus cells

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 58, No 3 (2020)

Article type

Original paper

Pages

227-234

Published online

2020-09-16

DOI

10.5603/FHC.a2020.0019

Pubmed

32936927

Bibliographic record

Folia Histochem Cytobiol 2020;58(3):227-234.

Keywords

intervertebral disc degeneration
rhTSG-6
extracellular matrix degradation
apoptosis
IL-1β
nucleus pulposus cells

Authors

Shishen Pei
Jinwei Ying
Yan Zhang
Linhao Su
Shi Cheng
Dike Ruan

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