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Ferrostatin-1 Inhibits Toll-Like Receptor 4/NF-κB Signaling to Alleviate Intervertebral Disc Degeneration in Rats

Published:January 19, 2023DOI:https://doi.org/10.1016/j.ajpath.2022.12.014
      Ferrostatin-1 (Fer-1) is an inhibitor of ferroptosis and has been documented to be implicated in the development of intervertebral disc degeneration (IDD). This study intends to explore the role of Fer-1 in IDD via the toll-like receptor 4 (TLR4)/NF-κB signaling pathway. Through the Gene Expression Omnibus database, IDD-related gene expression microarray GSE124272 and high-throughput sequencing data set GSE175710 were obtained. Then, differentially expressed genes in IDD were identified, followed by implementation of protein-protein interaction network analysis and receiver operating characteristic curve analysis. Main pathways in IDD were obtained through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes functional analyses, and target genes of Fer-1 were obtained through PubChem and PharmMapper websites. Finally, GPX4, FTH, and TLR4 expression was determined in a constructed IDD rat model. Three key co-expression modules involved in IDD were obtained through Weighted Gene Co-Expression Network Analysis. Thirteen differentially expressed genes were found to be associated with IDD, and eight key genes (TLR4, BCL2A1, CXCL1, IL1R1, NAMPT, SOCS3, XCL1, and IRAK3) were found to affect IDD. These eight key genes had the diagnostic potential for IDD. The NF-κB signaling pathway played a predominant role in IDD development. From network pharmacologic analysis, Fer-1 might suppress ferroptosis and ameliorate IDD via the TLR4/NF-κB signaling pathway, and the in vivo animal experiment further verified it. Fer-1 down-regulates TLR4 to inactivate NF-κB signaling pathway, suppressing ferroptosis and finally alleviating IDD in rats.

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      References

        • Kos N.
        • Gradisnik L.
        • Velnar T.
        A brief review of the degenerative intervertebral disc disease.
        Med Arch. 2019; 73: 421-424
        • Vergroesen P.P.
        • Kingma I.
        • Emanuel K.S.
        • Hoogendoorn R.J.
        • Welting T.J.
        • van Royen B.J.
        • van Dieen J.H.
        • Smit T.H.
        Mechanics and biology in intervertebral disc degeneration: a vicious circle.
        Osteoarthritis Cartilage. 2015; 23: 1057-1070
        • Dixon S.J.
        • Lemberg K.M.
        • Lamprecht M.R.
        • Skouta R.
        • Zaitsev E.M.
        • Gleason C.E.
        • Patel D.N.
        • Bauer A.J.
        • Cantley A.M.
        • Yang W.S.
        • Morrison 3rd, B.
        • Stockwell B.R.
        Ferroptosis: an iron-dependent form of nonapoptotic cell death.
        Cell. 2012; 149: 1060-1072
        • Zhang Y.
        • Han S.
        • Kong M.
        • Tu Q.
        • Zhang L.
        • Ma X.
        Single-cell RNA-seq analysis identifies unique chondrocyte subsets and reveals involvement of ferroptosis in human intervertebral disc degeneration.
        Osteoarthritis Cartilage. 2021; 29: 1324-1334
        • Lu S.
        • Song Y.
        • Luo R.
        • Li S.
        • Li G.
        • Wang K.
        • Liao Z.
        • Wang B.
        • Ke W.
        • Xiang Q.
        • Chen C.
        • Wu X.
        • Zhang Y.
        • Ling L.
        • Yang C.
        Ferroportin-dependent iron homeostasis protects against oxidative stress-induced nucleus pulposus cell ferroptosis and ameliorates intervertebral disc degeneration in vivo.
        Oxid Med Cell Longev. 2021; 2021: 6670497
        • Horwath M.C.
        • Bell-Horwath T.R.
        • Lescano V.
        • Krishnan K.
        • Merino E.J.
        • Deepe Jr., G.S.
        Antifungal activity of the lipophilic antioxidant ferrostatin-1.
        Chembiochem. 2017; 18: 2069-2078
        • Yang R.Z.
        • Xu W.N.
        • Zheng H.L.
        • Zheng X.F.
        • Li B.
        • Jiang L.S.
        • Jiang S.D.
        Involvement of oxidative stress-induced annulus fibrosus cell and nucleus pulposus cell ferroptosis in intervertebral disc degeneration pathogenesis.
        J Cell Physiol. 2021; 236: 2725-2739
        • Ge H.
        • Xue X.
        • Xian J.
        • Yuan L.
        • Wang L.
        • Zou Y.
        • Zhong J.
        • Jiang Z.
        • Shi J.
        • Chen T.
        • Su H.
        • Feng H.
        • Hu S.
        Ferrostatin-1 alleviates white matter injury via decreasing ferroptosis following spinal cord injury.
        Mol Neurobiol. 2022; 59: 161-176
        • Xiao Z.
        • Kong B.
        • Fang J.
        • Qin T.
        • Dai C.
        • Shuai W.
        • Huang H.
        Ferrostatin-1 alleviates lipopolysaccharide-induced cardiac dysfunction.
        Bioengineered. 2021; 12: 9367-9376
        • Shang P.
        • Tang Q.
        • Hu Z.
        • Huang S.
        • Hu Y.
        • Zhu J.
        • Liu H.
        Procyanidin B3 alleviates intervertebral disc degeneration via interaction with the TLR4/MD-2 complex.
        J Cell Mol Med. 2020; 24: 3701-3711
        • Shao Z.
        • Wang B.
        • Shi Y.
        • Xie C.
        • Huang C.
        • Chen B.
        • Zhang H.
        • Zeng G.
        • Liang H.
        • Wu Y.
        • Zhou Y.
        • Tian N.
        • Wu A.
        • Gao W.
        • Wang X.
        • Zhang X.
        Senolytic agent quercetin ameliorates intervertebral disc degeneration via the Nrf2/NF-kappaB axis.
        Osteoarthritis Cartilage. 2021; 29: 413-422
        • Chen X.
        • Xu S.
        • Zhao C.
        • Liu B.
        Role of TLR4/NADPH oxidase 4 pathway in promoting cell death through autophagy and ferroptosis during heart failure.
        Biochem Biophys Res Commun. 2019; 516: 37-43
        • Yang L.
        • Wang H.
        • Yang X.
        • Wu Q.
        • An P.
        • Jin X.
        • Liu W.
        • Huang X.
        • Li Y.
        • Yan S.
        • Shen S.
        • Liang T.
        • Min J.
        • Wang F.
        Auranofin mitigates systemic iron overload and induces ferroptosis via distinct mechanisms.
        Signal Transduct Target Ther. 2020; 5: 138
        • Clough E.
        • Barrett T.
        The gene expression omnibus database.
        Methods Mol Biol. 2016; 1418: 93-110
        • Wang Y.
        • Dai G.
        • Jiang L.
        • Liao S.
        • Xia J.
        Microarray analysis reveals an inflammatory transcriptomic signature in peripheral blood for sciatica.
        BMC Neurol. 2021; 21: 50
        • Ding S.L.
        • Zhang T.W.
        • Zhang Q.C.
        • Ding W.
        • Li Z.F.
        • Han G.J.
        • Bai J.S.
        • Li X.L.
        • Dong J.
        • Wang H.R.
        • Jiang L.B.
        Excessive mechanical strain accelerates intervertebral disc degeneration by disrupting intrinsic circadian rhythm.
        Exp Mol Med. 2021; 53: 1911-1923
        • Langfelder P.
        • Horvath S.
        WGCNA: an R package for weighted correlation network analysis.
        BMC Bioinformatics. 2008; 9: 559
        • Ritchie M.E.
        • Phipson B.
        • Wu D.
        • Hu Y.
        • Law C.W.
        • Shi W.
        • Smyth G.K.
        limma Powers differential expression analyses for RNA-sequencing and microarray studies.
        Nucleic Acids Res. 2015; 43: e47
        • Bardou P.
        • Mariette J.
        • Escudie F.
        • Djemiel C.
        • Klopp C.
        jvenn: An interactive Venn diagram viewer.
        BMC Bioinformatics. 2014; 15: 293
        • Szklarczyk D.
        • Gable A.L.
        • Lyon D.
        • Junge A.
        • Wyder S.
        • Huerta-Cepas J.
        • Simonovic M.
        • Doncheva N.T.
        • Morris J.H.
        • Bork P.
        • Jensen L.J.
        • Mering C.V.
        STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets.
        Nucleic Acids Res. 2019; 47: D607-D613
        • Zhu M.
        • Ye M.
        • Wang J.
        • Ye L.
        • Jin M.
        Construction of potential miRNA-mRNA regulatory network in COPD plasma by bioinformatics analysis.
        Int J Chron Obstruct Pulmon Dis. 2020; 15: 2135-2145
        • Kim S.
        • Chen J.
        • Cheng T.
        • Gindulyte A.
        • He J.
        • He S.
        • Li Q.
        • Shoemaker B.A.
        • Thiessen P.A.
        • Yu B.
        • Zaslavsky L.
        • Zhang J.
        • Bolton E.E.
        PubChem in 2021: new data content and improved web interfaces.
        Nucleic Acids Res. 2021; 49: D1388-D1395
        • Wang X.
        • Shen Y.
        • Wang S.
        • Li S.
        • Zhang W.
        • Liu X.
        • Lai L.
        • Pei J.
        • Li H.
        PharmMapper 2017 update: a web server for potential drug target identification with a comprehensive target pharmacophore database.
        Nucleic Acids Res. 2017; 45: W356-W360
        • Ji M.L.
        • Jiang H.
        • Zhang X.J.
        • Shi P.L.
        • Li C.
        • Wu H.
        • Wu X.T.
        • Wang Y.T.
        • Wang C.
        • Lu J.
        Preclinical development of a microRNA-based therapy for intervertebral disc degeneration.
        Nat Commun. 2018; 9: 5051
        • Liu H.
        • Cheng F.
        • Xu Q.
        • Huang W.
        • Wang S.
        • Sun R.
        • Ye D.
        • Zhang D.
        Lipoxin A4 suppresses angiotensin II type 1 receptor autoantibody in preeclampsia via modulating caspase-1.
        Cell Death Dis. 2020; 11: 78
        • Liu Y.
        • Zhou W.
        • Chen F.F.
        • Xiao F.
        • Zhu H.Y.
        • Zhou Y.
        • Guo G.C.
        Overexpression of LMP-1 decreases apoptosis in human nucleus pulposus cells via suppressing the NF-kappaB signaling pathway.
        Oxid Med Cell Longev. 2020; 2020: 8189706
        • Zhang X.
        • Huang Z.
        • Xie Z.
        • Chen Y.
        • Zheng Z.
        • Wei X.
        • Huang B.
        • Shan Z.
        • Liu J.
        • Fan S.
        • Chen J.
        • Zhao F.
        Homocysteine induces oxidative stress and ferroptosis of nucleus pulposus via enhancing methylation of GPX4.
        Free Radic Biol Med. 2020; 160: 552-565
        • Dangol S.
        • Chen Y.
        • Hwang B.K.
        • Jwa N.S.
        Iron- and reactive oxygen species-dependent ferroptotic cell death in rice-magnaporthe oryzae interactions.
        Plant Cell. 2019; 31: 189-209
        • Xu M.
        • Tao J.
        • Yang Y.
        • Tan S.
        • Liu H.
        • Jiang J.
        • Zheng F.
        • Wu B.
        Ferroptosis involves in intestinal epithelial cell death in ulcerative colitis.
        Cell Death Dis. 2020; 11: 86
        • Ohnishi T.
        • Iwasaki N.
        • Sudo H.
        Causes of and molecular targets for the treatment of intervertebral disc degeneration: a review.
        Cells. 2022; 11: 394
        • Shen L.
        • Xiao Y.
        • Wu Q.
        • Liu L.
        • Zhang C.
        • Pan X.
        TLR4/NF-kappaB axis signaling pathway-dependent up-regulation of miR-625-5p contributes to human intervertebral disc degeneration by targeting COL1A1.
        Am J Transl Res. 2019; 11: 1374-1388
        • Bi F.
        • Liu W.
        • Wu Z.
        • Ji C.
        • Chang C.
        Antiaging factor Klotho retards the progress of intervertebral disc degeneration through the toll-like receptor 4-NF-kappaB pathway.
        Int J Cell Biol. 2020; 2020: 8319516
        • Krock E.
        • Millecamps M.
        • Currie J.B.
        • Stone L.S.
        • Haglund L.
        Low back pain and disc degeneration are decreased following chronic toll-like receptor 4 inhibition in a mouse model.
        Osteoarthritis Cartilage. 2018; 26: 1236-1246
        • Guerrero-Hue M.
        • Garcia-Caballero C.
        • Palomino-Antolin A.
        • Rubio-Navarro A.
        • Vazquez-Carballo C.
        • Herencia C.
        • Martin-Sanchez D.
        • Farre-Alins V.
        • Egea J.
        • Cannata P.
        • Praga M.
        • Ortiz A.
        • Egido J.
        • Sanz A.B.
        • Moreno J.A.
        Curcumin reduces renal damage associated with rhabdomyolysis by decreasing ferroptosis-mediated cell death.
        FASEB J. 2019; 33: 8961-8975
        • Li S.
        • He Y.
        • Chen K.
        • Sun J.
        • Zhang L.
        • He Y.
        • Yu H.
        • Li Q.
        RSL3 drives ferroptosis through NF-kappaB pathway activation and GPX4 depletion in glioblastoma.
        Oxid Med Cell Longev. 2021; 2021: 2915019
        • Zhu K.
        • Zhu X.
        • Sun S.
        • Yang W.
        • Liu S.
        • Tang Z.
        • Zhang R.
        • Li J.
        • Shen T.
        • Hei M.
        Inhibition of TLR4 prevents hippocampal hypoxic-ischemic injury by regulating ferroptosis in neonatal rats.
        Exp Neurol. 2021; 345: 113828
        • Miotto G.
        • Rossetto M.
        • Di Paolo M.L.
        • Orian L.
        • Venerando R.
        • Roveri A.
        • Vuckovic A.M.
        • Bosello Travain V.
        • Zaccarin M.
        • Zennaro L.
        • Maiorino M.
        • Toppo S.
        • Ursini F.
        • Cozza G.
        Insight into the mechanism of ferroptosis inhibition by ferrostatin-1.
        Redox Biol. 2020; 28: 101328
        • Wei N.
        • Lu T.
        • Yang L.
        • Dong Y.
        • Liu X.
        Lipoxin A4 protects primary spinal cord neurons from Erastin-induced ferroptosis by activating the Akt/Nrf2/HO-1 signaling pathway.
        FEBS Open Bio. 2021; 11: 2118-2126
        • Luo Y.
        • Gao X.
        • Zou L.
        • Lei M.
        • Feng J.
        • Hu Z.
        Bavachin induces ferroptosis through the STAT3/P53/SLC7A11 axis in osteosarcoma cells.
        Oxid Med Cell Longev. 2021; 2021: 1783485
        • Xu Y.
        • Sang W.
        • Zhong Y.
        • Xue S.
        • Yang M.
        • Wang C.
        • Lu H.
        • Huan R.
        • Mao X.
        • Zhu L.
        • He C.
        • Ma J.
        CoCrMo-nanoparticles induced peri-implant osteolysis by promoting osteoblast ferroptosis via regulating Nrf2-ARE signalling pathway.
        Cell Prolif. 2021; 54: e13142
        • Tang W.
        • Guo J.
        • Liu W.
        • Ma J.
        • Xu G.
        Ferrostatin-1 attenuates ferroptosis and protects the retina against light-induced retinal degeneration.
        Biochem Biophys Res Commun. 2021; 548: 27-34
        • Tomalka J.A.
        • de Jesus T.J.
        • Ramakrishnan P.
        Sam68 is a regulator of toll-like receptor signaling.
        Cell Mol Immunol. 2017; 14: 107-117