Summary
Objective
Cartilaginous endplate (CEP) degeneration is the main early manifestations of intervertebral
disc degeneration (IVDD), and is closely related to the oxidative stress. Nrf2 (nuclear
factor E2-related factor 2, NFE2L2) is a vital transcriptional factor of cellular
antioxidant and anti-inflammatory responses. We aimed to illustrate whether the Nrf2
which was increased in expression by 4-octyl itaconate (4OI) could attenuate intervertebral
disc degeneration through suppressing macrophage associated inflammation and catabolism
of cartilaginous endplate.
Methods
Firstly, we detected the expression of Nrf2 in human degenerative CEPs. Then, we performed
in vitro, ex vivo and in vivo (a rat-tail puncture model) experiments to explore the role of 4OI in IVDD. Also,
by cell co-culture experiments, we demonstrated 4OI restrained the macrophage-associated
inflammatory responses. Finally, through western blotting and immunoprecipitation
(IP) assay, we clarified the ZNF598-mediated ubiquitination of Nrf2.
Results
We found decreased expression of Nrf2 in human degenerative CEPs. Using a rat IVDD
model(n = 6), 4OI significantly ameliorated the progression of IVDD by MR images and histological
analysis. Immunofluorescence results reveal that catabolism of CEPs and macrophage-associated
inflammation are suppressed by 4OI treatment. Mechanistically, the 4OI increases Nrf2
expression and inhibits the secretion of inflammatory factors (IL-1β) by Lipopolysaccharide
(LPS)-induced macrophages, thus preventing the inflammatory-related CEP degeneration.
Meanwhile, 4OI suppresses the reactive oxygen species (ROS) production and catabolism
of LPS-induced rat CEP cells. In addition, 4OI inhibits the ZNF598-dependent ubiquitination
of Nrf2 in LPS-induced rat CEP cells.
Conclusions
4OI may alleviate IVDD by suppressing CEP degeneration and macrophage-associated inflammation.
4OI may be an alternative therapy for degenerative CEPs/IVDs.
Keywords
Abbreviations:
LBP (low back pain), CEP (cartilaginous endplate), NP (nucleus pulposus), IVDD (Intervertebral disc degeneration), MMP (matrix metalloprotease), ECM (extracellular matrix), NFE2L2 (nuclear factor E2-related factor 2), 4OI (4-octyl itaconate), MitoSOX (mitochondrial superoxide), IL-1β (interleukin 1β), TNF-α (tumor necrosis factor α)To read this article in full you will need to make a payment
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References
- Will this patient develop persistent disabling low back pain?.JAMA. 2010; 303: 1295-1302
- Low back pain.N Engl J Med. 2001; 344: 363-370
- Lumbar disc disorders and low-back pain: socioeconomic factors and consequences.J Bone Joint Surg Am. 2006; 88: 21
- Discogenic origins of spinal instability.Spine. 2005; 30: 2621-2630https://doi.org/10.1097/01.brs.0000188203.71182.c0
- Intervertebral disc regeneration: From cell therapy to the development of novel bioinspired endogenous repair strategies.Adv Drug Deliv Rev. 2019; 146: 306-324https://doi.org/10.1016/j.addr.2018.04.017
- ISSLS prize winner: dynamic loading-induced convective transport enhances intervertebral disc nutrition.Spine. 2015; 40: 1158-1164https://doi.org/10.1097/Brs.0000000000001012
- On the mechanical behaviour of intervertebral discs.Spine. 1983; 8: 151-165
- Biomechanical and endplate effects on nutrient transport in the intervertebral disc.World Neurosurg. 2017; 99: 395-402https://doi.org/10.1016/j.wneu.2016.12.041
- 2001 Volvo Award Winner in Basic Science Studies: effect of nutrient supply on the viability of cells from the nucleus pulposus of the intervertebral disc.Spine. 2001; 26: 2543-2549
- Nutrition of the intervertebral disc.Spine (Phila Pa 1976). 2004; 29: 2700-2709
- Oxidative stress and exceptional human longevity: systematic review.Free Radic Biol Med. 2020; 149: 51-63https://doi.org/10.1016/j.freeradbiomed.2019.09.019
- ROS: crucial intermediators in the pathogenesis of intervertebral disc degeneration.Oxid Med Cell Longev. 2017; 2017: 1-12https://doi.org/10.1155/2017/5601593
- Oxidative/nitrosative stress in patients with modic changes: preliminary controlled study.Spine. 2015; 40: 1101-1107https://doi.org/10.1097/BRS.0000000000000737
- Autophagy is a protective response to the oxidative damage to endplate chondrocytes in intervertebral disc: implications for the treatment of degenerative lumbar disc.Oxid Med Cell Longev. 2017; 2017: 4041768https://doi.org/10.1155/2017/4041768
- Excessive reactive oxygen species are therapeutic targets for intervertebral disc degeneration.Arthritis Res Ther. 2015; 17: 316https://doi.org/10.1186/s13075-015-0834-8
- Small molecule inhibitor of TAK1 ameliorates rat cartilaginous endplate degeneration induced by oxidative stress in vitro and in vivo.Free Radic Biol Med. 2020; 148: 140-150https://doi.org/10.1016/j.freeradbiomed.2020.01.002
- The pore-forming protein gasdermin D regulates interleukin-1 secretion from living macrophages.Immunity. 2018; 48: 35-44.e36https://doi.org/10.1016/j.immuni.2017.11.013
- Inflammatory processes associated with canine intervertebral disc herniation.Front Immunol. 2017; 8: 1681https://doi.org/10.3389/fimmu.2017.01681
- Proinflammatory macrophages promote degenerative phenotypes in rat nucleus pulpous cells partly through ERK and JNK signaling.J Cell Physiol. 2019; 234: 5362-5371https://doi.org/10.1002/jcp.27507
- Alpha 2-macroglobulin as dual regulator for both anabolism and catabolism in the cartilaginous endplate of intervertebral disc.Spine. 2019; 44: E338-E347https://doi.org/10.1097/BRS.0000000000002852
- Upregulation of tumor necrosis factor alpha and ADAMTS-5, but not ADAMTS-4, in human intervertebral cartilage endplate with modic changes.Spine. 2014; 39: E817-E825https://doi.org/10.1097/BRS.0000000000000362
- Cartilaginous endplate avulsion is associated with modic changes and endplate defects, and residual back and leg pain following lumbar discectomy.Osteoarthr Cartil. 2021; 29: 707-717https://doi.org/10.1016/j.joca.2021.01.010
- Osteal tissue macrophages are involved in endplate osteosclerosis through the OSM-STAT3/YAP1 signaling axis in modic changes.J Immunol. 2020; 205: 968-980https://doi.org/10.4049/jimmunol.1901001
- Secreted factors from intervertebral disc cells and infiltrating macrophages promote degenerated intervertebral disc catabolism.Spine. 2019; 44: E520-E529https://doi.org/10.1097/BRS.0000000000002953
- NLRP3 inflammasome: from a danger signal sensor to a regulatory node of oxidative stress and inflammatory diseases.Redox Biol. 2015; 4: 296-307https://doi.org/10.1016/j.redox.2015.01.008
- Activation of the NRF2 antioxidant program sensitizes tumors to G6PD inhibition.Sci Adv. 2021; 7eabk1023https://doi.org/10.1126/sciadv.abk1023
- Senolytic agent Quercetin ameliorates intervertebral disc degeneration via the Nrf2/NF-kappaB axis.Osteoarthr Cartil. 2021; 29: 413-422https://doi.org/10.1016/j.joca.2020.11.006
- Itaconic acid is a mammalian metabolite induced during macrophage activation.J Am Chem Soc. 2011; 133: 16386-16389https://doi.org/10.1021/ja2070889
- The role of itaconate in host defense and inflammation.J Clin Investig. 2022; 132: e148548https://doi.org/10.1172/JCI148548
- Itaconate links inhibition of succinate dehydrogenase with macrophage metabolic remodeling and regulation of inflammation.Cell Metabol. 2016; 24: 158-166https://doi.org/10.1016/j.cmet.2016.06.004
- Itaconate is an anti-inflammatory metabolite that activates Nrf2 via alkylation of KEAP1.Nature. 2018; 556: 113-117https://doi.org/10.1038/nature25986
- A simple disc degeneration model induced by percutaneous needle puncture in the rat tail.Spine. 2008; 33: 1925-1934https://doi.org/10.1097/BRS.0b013e31817c64a9
- Pseurotin A inhibits osteoclastogenesis and prevents ovariectomized-induced bone loss by suppressing reactive oxygen species.Theranostics. 2019; 9: 1634-1650https://doi.org/10.7150/thno.30206
- The RCAN1.4-calcineurin/NFAT signaling pathway is essential for hypoxic adaption of intervertebral discs.Exp Mol Med. 2020; 52: 865-875https://doi.org/10.1038/s12276-020-0441-x
- Melatonin alleviates intervertebral disc degeneration by disrupting the IL-1β/NF-κB-NLRP3 inflammasome positive feedback loop.Bone Res. 2020; 8https://doi.org/10.1038/s41413-020-0087-2
- p16(Ink4a) deletion in cells of the intervertebral disc affects their matrix homeostasis and senescence associated secretory phenotype without altering onset of senescence.Matrix Biol J Int Soc Matrix Biol. 2019; https://doi.org/10.1016/j.matbio.2019.02.004
- Lycopene alleviates disc degeneration under oxidative stress through the Nrf2 signaling pathway.Mol Cell Probes. 2020; 51: 101559https://doi.org/10.1016/j.mcp.2020.101559
- Nutrient supply and nucleus pulposus cell function: effects of the transport properties of the cartilage endplate and potential implications for intradiscal biologic therapy.Osteoarthr Cartil. 2019; 27: 956-964https://doi.org/10.1016/j.joca.2019.01.013
- Damage to the human lumbar cartilage endplate and its clinical implications.Journal Anat. 2021; 238: 338-348https://doi.org/10.1111/joa.13321
- Intervertebral disc regeneration: do nutrients lead the way?.Nat Rev Rheumatol. 2014; 10: 561-566https://doi.org/10.1038/nrrheum.2014.91
- Interference in the endplate nutritional pathway causes intervertebral disc degeneration in an immature porcine model.Int Orthopaed. 2014; 38: 1011-1017https://doi.org/10.1007/s00264-014-2319-9
- The influence of direct inoculation of propionibacterium acnes on modic changes in the spine: evidence from a rabbit model.J Bone Joint Surg Am. 2017; 99: 472-481https://doi.org/10.2106/JBJS.16.00146
- Rapamycin induced autophagy inhibits inflammation-mediated endplate degeneration by enhancing Nrf2/Keap1 signaling of cartilage endplate stem cells.Stem Cells. 2019; 37: 828-840https://doi.org/10.1002/stem.2999
- 4-Octyl itaconate inhibits aerobic glycolysis by targeting GAPDH to exert anti-inflammatory effects.Nature Commun. 2019; 10: 5091https://doi.org/10.1038/s41467-019-13078-5
- Intervertebral disc ageing and degeneration: the antiapoptotic effect of oestrogen.Ageing Res Rev. 2020; 57: 100978https://doi.org/10.1016/j.arr.2019.100978
- Spontaneous resorption of lumbar disc herniation is less likely when modic changes are present.Spine. 2014; 39: 736-744https://doi.org/10.1097/BRS.0000000000000259
- Single-cell transcriptome profiling reveals multicellular ecosystem of nucleus pulposus during degeneration progression.Adv Sci (Weinh). 2022; 9: e2103631https://doi.org/10.1002/advs.202103631
- The immunomodulatory metabolite itaconate modifies NLRP3 and inhibits inflammasome activation.Cell Metabol. 2020; 32: 468-478https://doi.org/10.1016/j.cmet.2020.07.016
- Octyl itaconate inhibits osteoclastogenesis by suppressing Hrd1 and activating Nrf2 signaling.FASEB J Off Publ Fed Am Soc Exp Biol. 2019; 33: 12929-12940https://doi.org/10.1096/fj.201900887RR
- A small molecule NRF2 activator BC-1901S ameliorates inflammation through DCAF1/NRF2 axis.Redox Biol. 2020; 32: 101485https://doi.org/10.1016/j.redox.2020.101485
- The Nrf2 activator RTA-408 attenuates osteoclastogenesis by inhibiting STING dependent NF-kappab signaling.Redox Biol. 2020; 28: 101309https://doi.org/10.1016/j.redox.2019.101309
- Melatonin restores neutrophil functions and prevents apoptosis amid dysfunctional glutathione redox system.J Pineal Res. 2020; : e12676https://doi.org/10.1111/jpi.12676
- Impact of NF-κB pathway on the apoptosis-inflammation-autophagy crosstalk in human degenerative nucleus pulposus cells.Aging (Albany NY). 2019; 11: 7294-7306https://doi.org/10.18632/aging.102266
- NF-kappaB in the regulation of epithelial homeostasis and inflammation.Cell Res. 2011; 21: 146-158https://doi.org/10.1038/cr.2010.175
Article info
Publication history
Published online: October 17, 2022
Accepted:
October 11,
2022
Received:
May 17,
2022
Identification
Copyright
© 2022 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.