Abstract
Background
Intervertebral disk degeneration (IDD) is a common orthopedic disorder, and nucleus pulposus cells (NPCs) serve to stabilize the intervertebral disk.
Objective
This study was implemented to explore the protective effect of endplate stem cells (EPSCs) on IDD. NPCs and EPSCs were isolated from rats and tert-Butyl peroxide (TBHP) was used to treat NPCs and simulate IDD. EPSC-derived conditioned medium (CM) was collected and mixed with a culture medium at different proportions, which was then used to treat NPCs. SOD assay was employed to examine the oxidative stress level in the NPCs. To assess apoptosis and caspase-3 activity, flow cytometry and western blots for cleaved PARP1 and BAX were carried out. The mRNA expression levels of different genes were detected by quantitative real-time polymerase chain reaction. Furthermore, the concentrations of amphiregulin (Areg) and inflammatory factors were measured with ELISA kits and cell signaling pathways were evaluated using western blotting.
Results
Following TBHP treatment, caspase-3 activity in the NPCs increased. However, exposing TBHP-induced NPCs to the EPSC-derived CM reduced oxidative stress, caspase-3 activity, cell apoptosis, fibrotic response, and cytokine production. Additionally, the EPSC-derived CM contained a higher concentration of Areg compared to that in the NPCs. Furthermore, Areg decreased oxidative stress, cell apoptosis, fibrosis, and inflammation via both PI3K/AKT and ERK1/2 signaling pathways.
Conclusion
ECSCs reduce NPC apoptosis, oxidative stress, production of inflammatory factors, and fibrotic response via the release of Areg. Moreover, Areg can exert these protective effects on NPCs via PI3K/AKT and ERK1/2 signaling pathways.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Allon AA et al (2010) Structured coculture of stem cells and disc cells prevent disc degeneration in a rat model. Spine J 10:1089–1097. https://doi.org/10.1016/j.spinee.2010.09.014
Bai Z et al (2020) Protective effects of autophagy and NFE2L2 on reactive oxygen species-induced pyroptosis of human nucleus pulposus cells. Aging (albany NY) 12:7534–7548. https://doi.org/10.18632/aging.103109
Benzakour T, Benzakour A (2019) Disc herniation and disc disease: the present and the future of management. Int Orthop 43:755–760. https://doi.org/10.1007/s00264-019-04324-y
Carnet O et al (2015) Mesenchymal stem cells shed amphiregulin at the surface of lung carcinoma cells in a juxtacrine manner. Neoplasia 17:552–563. https://doi.org/10.1016/j.neo.2015.07.002
Chen C et al (2020) Autologous fibroblasts induce fibrosis of the nucleus pulposus to maintain the stability of degenerative intervertebral discs. Bone Res 8:7. https://doi.org/10.1038/s41413-019-0082-7
Farshad-Amacker NA, Hughes A, Herzog RJ, Seifert B, Farshad M (2017) The intervertebral disc, the endplates and the vertebral bone marrow as a unit in the process of degeneration. Eur Radiol 27:2507–2520. https://doi.org/10.1007/s00330-016-4584-z
Feng C et al (2017) ROS: crucial intermediators in the pathogenesis of intervertebral disc degeneration. Oxid Med Cell Longev 2017:5601593. https://doi.org/10.1155/2017/5601593
Hu B et al (2018) Intervertebral disc-derived stem/progenitor cells as a promising cell source for intervertebral disc regeneration. Stem Cells Int 2018:7412304. https://doi.org/10.1155/2018/7412304
Ko JH, Kim HJ, Jeong HJ, Lee HJ, Oh JY (2020) Mesenchymal stem and stromal cells harness macrophage-derived amphiregulin to maintain tissue homeostasis. Cell Rep 30:3806-3820.e3806. https://doi.org/10.1016/j.celrep.2020.02.062
Leung VY et al (2014) Mesenchymal stem cells reduce intervertebral disc fibrosis and facilitate repair. Stem Cells 32:2164–2177. https://doi.org/10.1002/stem.1717
Luo L et al (2021) Cartilage endplate stem cells inhibit intervertebral disc degeneration by releasing exosomes to nucleus pulposus cells to activate Akt/autophagy. Stem Cells 39:467–481. https://doi.org/10.1002/stem.3322
Mahtouk K et al (2005) Expression of EGF-family receptors and amphiregulin in multiple myeloma. Amphiregulin is a growth factor for myeloma cells. Oncogene 24:3512–3524. https://doi.org/10.1038/sj.onc.1208536
Meng C et al (2020) Amphiregulin inhibits TNF-α-induced alveolar epithelial cell death through EGFR signaling pathway. Biomed Pharmacother 125:109995. https://doi.org/10.1016/j.biopha.2020.109995
Ogata-Suetsugu S et al (2017) Amphiregulin suppresses epithelial cell apoptosis in lipopolysaccharide-induced lung injury in mice. Biochem Biophys Res Commun 484:422–428. https://doi.org/10.1016/j.bbrc.2017.01.142
Oichi T, Taniguchi Y, Oshima Y, Tanaka S, Saito T (2020) Pathomechanism of intervertebral disc degeneration. JOR Spine 3:e1076. https://doi.org/10.1002/jsp2.1076
Pan T et al (2018) Geniposide suppresses interleukin-1β-induced inflammation and apoptosis in rat chondrocytes via the PI3K/Akt/NF-κB signaling pathway. Inflammation 41:390–399. https://doi.org/10.1007/s10753-017-0694-2
Pereira CL et al (2016) Mesenchymal stem/stromal cells seeded on cartilaginous endplates promote intervertebral disc regeneration through extracellular matrix remodeling. Sci Rep 6:33836. https://doi.org/10.1038/srep33836
Platen C et al (2019) Amphiregulin regulates phagocytosis-induced cell death in monocytes via EGFR and the Bcl-2 protein family. Mediators Inflamm 2019:1603131. https://doi.org/10.1155/2019/1603131
Ravindra VM et al (2018) Degenerative lumbar spine disease: estimating global incidence and worldwide volume. Glob Spine J 8:784–794. https://doi.org/10.1177/2192568218770769
Risbud MV, Shapiro IM (2014) Role of cytokines in intervertebral disc degeneration: pain and disc content. Nat Rev Rheumatol 10:44–56. https://doi.org/10.1038/nrrheum.2013.160
Teraguchi M et al (2017) Progression, incidence, and risk factors for intervertebral disc degeneration in a longitudinal population-based cohort: the Wakayama Spine Study. Osteoarthr Cartil 25:1122–1131. https://doi.org/10.1016/j.joca.2017.01.001
Wang Y, Jiang L, Dai G, Li S, Mu X (2018) Bioinformatics analysis reveals different gene expression patterns in the annulus fibrosis and nucleus pulpous during intervertebral disc degeneration. Exp Ther Med 16:5031–5040. https://doi.org/10.3892/etm.2018.6884
Wang H, Jiang Z, Pang Z, Zhou T, Gu Y (2020a) Acacetin alleviates inflammation and matrix degradation in nucleus pulposus cells and ameliorates intervertebral disc degeneration in vivo. Drug Des Dev Ther 14:4801–4813. https://doi.org/10.2147/dddt.S274812
Wang Z, Sun W, Sun X, Wang Y, Zhou M (2020b) Kaempferol ameliorates Cisplatin induced nephrotoxicity by modulating oxidative stress, inflammation and apoptosis via ERK and NF-κB pathways. AMB Express 10:58. https://doi.org/10.1186/s13568-020-00993-w
Willmarth NE et al (2009) Altered EGFR localization and degradation in human breast cancer cells with an amphiregulin/EGFR autocrine loop. Cell Signal 21:212–219. https://doi.org/10.1016/j.cellsig.2008.10.003
Wu Y, Wang Z, Fu X, Lin Z, Yu K (2020) Geraniol-mediated osteoarthritis improvement by down-regulating PI3K/Akt/NF-κB and MAPK signals: In vivo and in vitro studies. Int Immunopharmacol 86:106713. https://doi.org/10.1016/j.intimp.2020.106713
Xia K et al (2019) Differentiation of pluripotent stem cells into nucleus pulposus progenitor cells for intervertebral disc regeneration. Curr Stem Cell Res Ther 14:57–64. https://doi.org/10.2174/1574888x13666180918095121
Yang F, Leung VY, Luk KD, Chan D, Cheung KM (2009) Mesenchymal stem cells arrest intervertebral disc degeneration through chondrocytic differentiation and stimulation of endogenous cells. Mol Ther 17:1959–1966. https://doi.org/10.1038/mt.2009.146
Yang F, Xu W, Pei Y (2021) Amphiregulin induces interleukin-8 production and cell proliferation in lung epithelial cells through PI3K-Akt/ERK pathways. Eur J Inflam 19:2058739221998202
Yao Y et al (2016) Data for the gene expression profiling and alternative splicing events during the chondrogenic differentiation of human cartilage endplate-derived stem cells under hypoxia. Data Brief 7:1438–1442. https://doi.org/10.1016/j.dib.2016.04.037
Yotsumoto F et al (2010) Amphiregulin regulates the activation of ERK and Akt through epidermal growth factor receptor and HER3 signals involved in the progression of pancreatic cancer. Cancer Sci 101:2351–2360. https://doi.org/10.1111/j.1349-7006.2010.01671.x
Yu Y et al (2019) Amphiregulin promotes trophoblast invasion and increases MMP9/TIMP1 ratio through ERK1/2 and Akt signal pathways. Life Sci 236:116899. https://doi.org/10.1016/j.lfs.2019.116899
Zhang Z et al (2020) Klotho alleviates lung injury caused by paraquat via suppressing ROS/P38 MAPK-regulated inflammatory responses and apoptosis. Oxid Med Cell Longev 2020:1854206. https://doi.org/10.1155/2020/1854206
Zhao R, Liu W, Xia T, Yang L (2019) Disordered mechanical stress and tissue engineering therapies in intervertebral disc degeneration. Polymers (basel). https://doi.org/10.3390/polym11071151
Zheng J et al (2021) Reactive oxygen species mediate low back pain by upregulating substance p in intervertebral disc degeneration. Oxid Med Cell Longev 2021:6681815. https://doi.org/10.1155/2021/6681815
Zuo WL et al (2017) EGF-Amphiregulin interplay in airway stem/progenitor cells links the pathogenesis of smoking-induced lesions in the human airway epithelium. Stem Cells 35:824–837. https://doi.org/10.1002/stem.2512
Acknowledgements
This work was supported by Youth Science Foundation of Jiangxi Province (No.20171BAB215026) and Natural Science Foundation of Jiangxi Province (No.20192BAB205029).
Author information
Authors and Affiliations
Contributions
QC, YW and NL conceived the study and designed the experiments. MZ, CX and RC contributed to the data collection, performed the data analysis and interpreted the results. QC and YW wrote the manuscript; Ning Liu contributed to the critical revision of article. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
Author Qin Chen declares that she has no conflict of interest. Author Yaohong Wu declares that she has no conflict of interest. Author Mingliang Zhong declares that she has no conflict of interest. Author Chanhua Xu declares that she has no conflict of interest. Author Rongchun Chen declares that she has no conflict of interest. Author Ning Liu declares that she has no conflict of interest.
Compliance with ethical standards section
Our animal experiments were approved by the Ethical Committee of Ganzhou People's Hospital and were performed under the guidelines published by the committee. Approval number: TY-HKY-2020-012-02.
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
Chen, Q., Wu, Y., Zhong, M. et al. Amphiregulin secreted by cartilage endplate stem cells inhibits intervertebral disk degeneration and TNF-α production via PI3K/AKT and ERK1/2 signaling pathways. Mol. Cell. Toxicol. 19, 255–264 (2023). https://doi.org/10.1007/s13273-022-00254-w
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13273-022-00254-w