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Rutin maintains redox balance to relieve oxidative stress induced by TBHP in nucleus pulposus cells

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Abstract

Rutin is well known for its anti-inflammatory and antioxidant properties against oxidative stress. However, its protective function in nucleus pulposus cells (NPCs) remains unclear. This study was aimed to explore the effects of rutin on oxidative stress in NPCs. Primary NPCs were obtained from 1-mo-old SD rats. The NPCs were treated with tert-butyl hydrogen peroxide (TBHP) to obtain the oxidative stress, and different concentrations of rutin were used to observe its influence on the oxidative stress in NPCs. Fluorescent probe DCFH-DA was used to detect reactive oxide species (ROS). The antioxidant proteins and genes of heat shock protein 70 (HSP70), manganese superoxide dismutase (Mn-SOD), catalase, aggrecan and collagen II in NPCs were measured by western blot and real-time PCR. With the stimulation of TBHP, the content of ROS in NPCs increased significantly and showed solubility correlation. Rutin effectively reduced the accumulation of ROS in a dose-dependent manner. The antioxidant proteins of HSP70, Mn-SOD, and catalase and the matrix proteins of aggrecan and collagen II decreased remarkably with the stimulation of TBHP, while the matrix metalloproteinase-13 (MMP-13) significantly increased after TBHP intervention. Rutin boosted the expressions of the HSP70, Mn-SOD, and catalase, elevated the contents of aggrecan and collagen II, and inhibited the expression of MMP-13 in NPCs. The findings of this study suggested that rutin is able to reverse oxidative stress and maintain cellular function of NPCs, and it was indicated that rutin could be a possible therapeutic option for intervertebral disc degeneration.

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References

  • Afanas'Ev IB, Dorozhko AI, Brodskii AV, Kostyuk VA, Potapovitch AI (1989) Chelating and free radical scavenging mechanisms of inhibitory action of rutin and quercetin in lipid peroxidation. Biochem Pharmacol 38(11):1763–1769

    Article  CAS  Google Scholar 

  • Alvarez-Garcia O, Matsuzaki T, Olmer M, Masuda K, Lotz MK (2017) Age-related reduction in the expression of FOXO transcription factors and correlations with intervertebral disc degeneration. J Orthop Res 35(12):2682–2691

    Article  CAS  Google Scholar 

  • Chen D, Xia D, Pan Z, Xu D, Zhou Y, Wu Y, Cai N, Tang Q, Wang C, Yan M, Zhang JJ, Zhou K, Wang Q, Feng Y, Wang X, Xu H, Zhang X, Tian N (2016) Metformin protects against apoptosis and senescence in nucleus pulposus cells and ameliorates disc degeneration in vivo. Cell Death Dis 7(10):e2441

    Article  CAS  Google Scholar 

  • Dimozi A, Mavrogonatou E, Sklirou A, Kletsas D (2015) Oxidative stress inhibits the proliferation, induces premature senescence and promotes a catabolic phenotype in human nucleus pulposus intervertebral disc cells. Eur Cell Mater 30(89-102):103

    Google Scholar 

  • Feng C, Yang M, Lan M, Liu C, Zhang Y, Huang B et al (2017) ROS: crucial intermediators in the pathogenesis of intervertebral disc degeneration. Oxidative Med Cell Longev 2017:5601593

    Google Scholar 

  • Ganeshpurkar A, Saluja AK (2017) The pharmacological potential of rutin. Saudi Pharm J 25(2):149–164

    Article  Google Scholar 

  • Gul A, Kunwar B, Mazhar M, Faizi S, Ahmed D, Shah MR, Simjee SU (2018) Rutin and rutin-conjugated gold nanoparticles ameliorate collagen-induced arthritis in rats through inhibition of NF-kappaB and iNOS activation. Int Immunopharmacol 59:310–317

    Article  CAS  Google Scholar 

  • Hou G, Lu H, Chen M, Yao H, Zhao H (2014) Oxidative stress participates in age-related changes in rat lumbar intervertebral discs. Arch Gerontol Geriatr 59(3):665–669

    Article  CAS  Google Scholar 

  • Kauppila T, Kauppila J, Larsson NG (2017) Mammalian mitochondria and aging: an update. Cell Metab 25(1):57–71

    Article  CAS  Google Scholar 

  • Khomenko IP, Bakhtina LY, Zelenina OM, Kruglov SV, Manukhina EB, Bayda LA, Malyshev IY (2007) Role of heat shock proteins HSP70 and HSP32 in the protective effect of adaptation of cultured HT22 hippocampal cells to oxidative stress. Bull Exp Biol Med 144(2):174–177

    Article  CAS  Google Scholar 

  • Kong G, Huang Z, Ji W, Wang X, Liu J, Wu X, Huang Z, Li R, Zhu Q (2017) The ketone metabolite beta-hydroxybutyrate attenuates oxidative stress in spinal cord injury by suppression of class I histone deacetylases. J Neurotrauma 34(18):2645–2655

    Article  Google Scholar 

  • Kurashova NA, Madaeva IM, Kolesnikova LI (2019) Expression of heat shock proteins HSP70 under oxidative stress. Adv Gerontol 32(4):502–508

    CAS  PubMed  Google Scholar 

  • Le Maitre CL, Pockert A, Buttle DJ, Freemont AJ, Hoyland JA (2007) Matrix synthesis and degradation in human intervertebral disc degeneration. Biochem Soc Trans 35(Pt 4):652–655

    PubMed  Google Scholar 

  • Magalingam KB, Radhakrishnan A, Haleagrahara N (2013) Rutin, a bioflavonoid antioxidant protects rat pheochromocytoma (PC-12) cells against 6-hydroxydopamine (6-OHDA)-induced neurotoxicity. Int J Mol Med 32(1):235–240

    Article  CAS  Google Scholar 

  • Martinez DTI, De la Fuente M (2015) The role of Hsp70 in oxi-inflamm-aging and its use as a potential biomarker of lifespan. Biogerontology 16(6):709–721

    Article  Google Scholar 

  • Na JY, Song K, Kim S, Kwon J (2016) Rutin protects rat articular chondrocytes against oxidative stress induced by hydrogen peroxide through SIRT1 activation. Biochem Biophys Res Commun 473(4):1301–1308

    Article  CAS  Google Scholar 

  • Nerlich AG, Bachmeier BE, Schleicher E, Rohrbach H, Paesold G, Boos N (2007) Immunomorphological analysis of RAGE receptor expression and NF-kappaB activation in tissue samples from normal and degenerated intervertebral discs of various ages. Ann N Y Acad Sci 1096:239–248

    Article  CAS  Google Scholar 

  • Nerlich AG, Schleicher ED, Boos N (1997) 1997 Volvo Award winner in basic science studies. Immunohistologic markers for age-related changes of human lumbar intervertebral discs. Spine (Phila Pa 1976) 22(24):2781–2795

    Article  CAS  Google Scholar 

  • Peng B, Hou S, Shi Q, Jia L (2001) The relationship between cartilage end-plate calcification and disc degeneration: an experimental study. Chin Med J 114(3):308–312

    CAS  PubMed  Google Scholar 

  • Pulickal T, Boos J, Konieczny M, Sawicki LM, Muller-Lutz A, Bittersohl B et al (2019) MRI identifies biochemical alterations of intervertebral discs in patients with low back pain and radiculopathy. Eur Radiol 29(12):6443–6446

    Article  Google Scholar 

  • Sabirzhanov B, Stoica BA, Hanscom M, Piao CS, Faden AI (2012) Over-expression of HSP70 attenuates caspase-dependent and caspase-independent pathways and inhibits neuronal apoptosis. J Neurochem 123(4):542–554

    Article  CAS  Google Scholar 

  • Sandy JD (2001) Proteoglycan core proteins and catabolic fragments present in tissues and fluids. Methods Mol Biol 171:335–345

    CAS  PubMed  Google Scholar 

  • Sivan SS, Tsitron E, Wachtel E, Roughley P, Sakkee N, van der Ham F, Degroot J, Maroudas A (2006) Age-related accumulation of pentosidine in aggrecan and collagen from normal and degenerate human intervertebral discs. Biochem J 399(1):29–35

    Article  CAS  Google Scholar 

  • Song K, Kim S, Na JY, Park JH, Kim JK, Kim JH, Kwon J (2014) Rutin attenuates ethanol-induced neurotoxicity in hippocampal neuronal cells by increasing aldehyde dehydrogenase 2. Food Chem Toxicol 72:228–233

    Article  CAS  Google Scholar 

  • Su KY, Yu CY, Chen YW, Huang YT, Chen CT, Wu HF, Chen YLS (2014) Rutin, a flavonoid and principal component of Saussurea involucrata, attenuates physical fatigue in a forced swimming mouse model. Int J Med Sci 11(5):528–537

    Article  CAS  Google Scholar 

  • Suzuki S, Fujita N, Hosogane N, Watanabe K, Ishii K, Toyama Y, Takubo K, Horiuchi K, Miyamoto T, Nakamura M, Matsumoto M (2015) Excessive reactive oxygen species are therapeutic targets for intervertebral disc degeneration. Arthritis Res Ther 17:316

    Article  Google Scholar 

  • Tonomura H, Takahashi KA, Mazda O, Arai Y, Shin-Ya M, Inoue A, Honjo K, Hojo T, Imanishi J, Kubo T (2008) Effects of heat stimulation via microwave applicator on cartilage matrix gene and HSP70 expression in the rabbit knee joint. J Orthop Res 26(1):34–41

    Article  CAS  Google Scholar 

  • Valko M, Jomova K, Rhodes CJ, Kuca K, Musilek K (2016) Redox- and non-redox-metal-induced formation of free radicals and their role in human disease. Arch Toxicol 90(1):1–37

    Article  CAS  Google Scholar 

  • Vo NV, Hartman RA, Patil PR, Risbud MV, Kletsas D, Iatridis JC, Hoyland JA, le Maitre CL, Sowa GA, Kang JD (2016) Molecular mechanisms of biological aging in intervertebral discs. J Orthop Res 34(8):1289–1306

    Article  Google Scholar 

  • Vo N, Niedernhofer LJ, Nasto LA, Jacobs L, Robbins PD, Kang J, Evans CH (2013) An overview of underlying causes and animal models for the study of age-related degenerative disorders of the spine and synovial joints. J Orthop Res 31(6):831–837

    Article  Google Scholar 

  • Yang D, Wang D, Shimer A, Shen FH, Li X, Yang X (2014) Glutathione protects human nucleus pulposus cells from cell apoptosis and inhibition of matrix synthesis. Connect Tissue Res 55(2):132–139

    Article  CAS  Google Scholar 

  • Yang L, Rong Z, Zeng M, Cao Y, Gong X, Lin L, Chen Y, Cao W, Zhu L, Dong W (2015) Pyrroloquinoline quinone protects nucleus pulposus cells from hydrogen peroxide-induced apoptosis by inhibiting the mitochondria-mediated pathway. Eur Spine J 24(8):1702–1710

    Article  Google Scholar 

  • Zeng Y, Song J, Zhang M, Wang H, Zhang Y, Suo H (2020) Comparison of in vitro and in vivo antioxidant activities of six flavonoids with similar structures. Antioxidants (Basel) 9(8)

  • Zhai X, Ding Y, Wang Q, Zhang H, Li F (2016) Rutin acid ameliorates neural apoptosis induced by traumatic brain injury via mitochondrial pathways in mice. Neuroimmunomodulation 23(3):179–187

    Article  CAS  Google Scholar 

  • Zhang S, Liu W, Wang P, Hu B, Lv X, Chen S, Wang B, Shao Z (2021) Activation of HSP70 impedes tert-butyl hydroperoxide (t-BHP)-induced apoptosis and senescence of human nucleus pulposus stem cells via inhibiting the JNK/c-Jun pathway. Mol Cell Biochem

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Funding

This article was supported by the Scientific Research Projects of Administration of Traditional Chinese Medicine of Guangdong (No. 20211243 and No. 20201231), the Basic and Applied Basic Research Fund Project of Guangdong Province (No. 2020A1515110631), the Health Science and Technology Project of Guangzhou (No. 20211A011078), the Medical Science and Technology Research Fund Project of Guangdong (No. 2020111220372206), and the Science and Technology Plan Project of Guangzhou (No. 201704020032).

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Author notes

  1. Jian Zhou and Qi Liu contributed equally to this work and should be considered co-first authors.

Authors and Affiliations

  1. Department of Spine and Joint Surgery, Nanchang Hongdu Hospital of TCM, Nanchang, Jiangxi, China

    Jian Zhou

  2. Department of Orthopaedic Surgery, The Second Affiliated Hospital of Guangzhou Medical University, 250 Chang-gang-dong Road, Guangzhou, 510260, Guangdong, China

    Qi Liu, Chuhai Xie, Long Ling, Hailan Hu, Yanming Cao & Yan Huang

  3. Department of Orthopaedic Surgery, Southern University of Science and Technology Hospital, Shenzhen, China

    Zhou Yang

  4. School of Traditional Chinese Medicine, Southern Medical University, 1063 Shatai Road, Guangzhou, 510515, Guangdong, China

    Yue Hua

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  1. Jian Zhou
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  2. Qi Liu
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  3. Zhou Yang
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  4. Chuhai Xie
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  5. Long Ling
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  7. Yanming Cao
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  8. Yan Huang
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  9. Yue Hua
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Contributions

QL and JZ were the major contributors in writing the manuscript and performing the experiments. QL and YH designed the study and were in charge of reviewing the manuscript. ZY, CX, and LL performed the experiments and analyzed the data. HH, YC, and YH interpreted the data and edited the important intellectual content of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Qi Liu or Yue Hua.

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All experiments were approved by the Institutional Animal Care and Use Committee of Guangzhou Medical University.

Competing interests

The authors declare no competing interests.

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Editor: Tetsuji Okamoto

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Zhou, J., Liu, Q., Yang, Z. et al. Rutin maintains redox balance to relieve oxidative stress induced by TBHP in nucleus pulposus cells. In Vitro Cell.Dev.Biol.-Animal 57, 448–456 (2021). https://doi.org/10.1007/s11626-021-00581-7

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  • DOI: https://doi.org/10.1007/s11626-021-00581-7

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