Baicalein Inhibits the IL-1β-Induced Inflammatory Response in Nucleus Pulposus Cells and Attenuates Disc Degeneration In vivo

Abstract

Intervertebral disc degeneration (IDD) is widely considered one of the main causes of low back pain, which is a chronic progressive disease closely related to inflammation and degeneration of nucleus pulposus (NP) cells. Baicalein is a natural bioactive compound with anti-inflammatory effects in different diseases, including inhibition of the inflammatory response in chondrocytes, whose morphology and avascular supply are similar to those of NP cells. Therefore, we hypothesized that baicalein may have a therapeutic effect on IDD by suppressing the inflammatory response. In vitro, NP cells were pretreated with baicalein for 2 h and then incubated with IL-1β for 24 h. We found that baicalein not only inhibited the overexpression of inflammatory cytokine production, including NO, PGE2, TNF-α, and IL-6, but also suppressed the expression of COX-2 and iNOS. The IL-1β-induced overexpression of MMP13 and ADAMTS5 and degradation of aggrecan and type II collagen were reversed by baicalein in a dose-dependent manner. Mechanistically, we found that baicalein suppressed the IL-1β-induced activation of the NF-κB and MAPK pathways. Moreover, an in vivo study demonstrated that baicalein treatment could ameliorate IDD in a puncture-induced rat model. Thus, baicalein has great value as a potential therapeutic agent for IDD.

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References

  1. 1.

    Katz, J.N. 2006. Lumbar disc disorders and low-back pain: socioeconomic factors and consequences. The Journal of bone and joint surgery American volume 88 (Suppl 2): 21–24.

    PubMed  Google Scholar 

  2. 2.

    Luoma, K., H. Riihimaki, R. Luukkonen, R. Raininko, E. Viikari-Juntura, and A. Lamminen. 2000. Low back pain in relation to lumbar disc degeneration. Spine 25: 487–492.

    CAS  PubMed  Google Scholar 

  3. 3.

    Adams, M.A., and P.J. Roughley. 2006. What is intervertebral disc degeneration, and what causes it? Spine 31: 2151–2161.

    PubMed  Google Scholar 

  4. 4.

    Hangai, M., K. Kaneoka, S. Kuno, S. Hinotsu, M. Sakane, N. Mamizuka, S. Sakai, and N. Ochiai. 2008. Factors associated with lumbar intervertebral disc degeneration in the elderly. The spine journal : official journal of the North American Spine Society 8: 732–740.

    Google Scholar 

  5. 5.

    Burke, J.G., R.W. Watson, D. McCormack, F.E. Dowling, M.G. Walsh, and J.M. Fitzpatrick. 2002. Intervertebral discs which cause low back pain secrete high levels of proinflammatory mediators. The Journal of bone and joint surgery British volume 84: 196–201.

    CAS  PubMed  Google Scholar 

  6. 6.

    Shamji, M.F., L.A. Setton, W. Jarvis, S. So, J. Chen, L. Jing, R. Bullock, R.E. Isaacs, C. Brown, and W.J. Richardson. 2010. Proinflammatory cytokine expression profile in degenerated and herniated human intervertebral disc tissues. Arthritis and Rheumatism 62: 1974–1982.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Le Maitre, C.L., A.J. Freemont, and J.A. Hoyland. 2005. The role of interleukin-1 in the pathogenesis of human intervertebral disc degeneration. Arthritis Research & Therapy 7: R732–R745.

    Google Scholar 

  8. 8.

    Lee, S., C.S. Moon, D. Sul, J. Lee, M. Bae, Y. Hong, M. Lee, S. Choi, R. Derby, B.J. Kim, J. Kim, J.S. Yoon, L. Wolfer, J. Kim, J. Wang, S.W. Hwang, and S.H. Lee. 2009. Comparison of growth factor and cytokine expression in patients with degenerated disc disease and herniated nucleus pulposus. Clinical Biochemistry 42: 1504–1511.

    CAS  PubMed  Google Scholar 

  9. 9.

    Wuertz, K., N. Vo, D. Kletsas, and N. Boos. 2012. Inflammatory and catabolic signalling in intervertebral discs: the roles of NF-kappaB and MAP kinases. European Cells & Materials 23: 103–119 discussion 119-120.

    CAS  Google Scholar 

  10. 10.

    Li, H.B., Y. Jiang, and F. Chen. 2004. Separation methods used for Scutellaria baicalensis active components. Journal of chromatography B, Analytical technologies in the biomedical and life sciences 812: 277–290.

    CAS  PubMed  Google Scholar 

  11. 11.

    Kim, K.C., I.K. Lee, K.A. Kang, H.S. Kim, S.S. Kang, and J.W. Hyun. 2012. Baicalein (5,6,7-trihydroxyflavone) reduces oxidative stress-induced DNA damage by upregulating the DNA repair system. Cell Biology and Toxicology 28: 421–433.

    CAS  PubMed  Google Scholar 

  12. 12.

    Zhu, J., J. Wang, Y. Sheng, Y. Zou, L. Bo, F. Wang, J. Lou, X. Fan, R. Bao, Y. Wu, F. Chen, X. Deng, and J. Li. 2012. Baicalin improves survival in a murine model of polymicrobial sepsis via suppressing inflammatory response and lymphocyte apoptosis. PLoS One 7: e35523.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Fan, G.W., Y. Zhang, X. Jiang, Y. Zhu, B. Wang, L. Su, W. Cao, H. Zhang, and X. Gao. 2013. Anti-inflammatory activity of baicalein in LPS-stimulated RAW264.7 macrophages via estrogen receptor and NF-kappaB-dependent pathways. Inflammation 36: 1584–1591.

    CAS  PubMed  Google Scholar 

  14. 14.

    Yang, W., H. Li, X. Cong, X. Wang, Z. Jiang, Q. Zhang, X. Qi, S. Gao, R. Cao, and W. Tian. 2016. Baicalin attenuates lipopolysaccharide induced inflammation and apoptosis of cow mammary epithelial cells by regulating NF-kappaB and HSP72. International Immunopharmacology 40: 139–145.

    CAS  PubMed  Google Scholar 

  15. 15.

    Dong, S.J., Y.Q. Zhong, W.T. Lu, G.H. Li, H.L. Jiang, and B. Mao. 2015. Baicalin inhibits lipopolysaccharide-induced inflammation through signaling NF-kappaB pathway in HBE16 airway epithelial cells. Inflammation 38: 1493–1501.

    CAS  PubMed  Google Scholar 

  16. 16.

    Zhang, X., Y. Yang, L. Du, W. Zhang, and G. Du. 2017. Baicalein exerts anti-neuroinflammatory effects to protect against rotenone-induced brain injury in rats. International Immunopharmacology 50: 38–47.

    CAS  PubMed  Google Scholar 

  17. 17.

    Zhang, X., Y. Zhu, X. Chen, Y. Zhang, Y. Zhang, Y. Jia, H. Wang, Y. Liu, and L. Xiao. 2014. Baicalein ameliorates inflammatory-related apoptotic and catabolic phenotypes in human chondrocytes. International Immunopharmacology 21: 301–308.

    PubMed  Google Scholar 

  18. 18.

    Chen, W.P., Y. Xiong, P.F. Hu, J.P. Bao, and L.D. Wu. 2015. Baicalein inhibits MMPs expression via a MAPK-dependent mechanism in chondrocytes. Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology 36: 325–333.

    Google Scholar 

  19. 19.

    Li, Y., J. Wang, X. Song, H. Bai, T. Ma, Z. Zhang, X. Li, R. Jiang, G. Wang, X. Fan, X. Liu, and L. Gao. 2017. Effects of baicalein on IL-1beta-induced inflammation and apoptosis in rat articular chondrocytes. Oncotarget 8: 90781–90795.

    PubMed  PubMed Central  Google Scholar 

  20. 20.

    Pfirrmann, C.W., A. Metzdorf, M. Zanetti, J. Hodler, and N. Boos. 2001. Magnetic resonance classification of lumbar intervertebral disc degeneration. Spine 26: 1873–1878.

    CAS  PubMed  Google Scholar 

  21. 21.

    Han, B., K. Zhu, F.C. Li, Y.X. Xiao, J. Feng, Z.L. Shi, M. Lin, J. Wang, and Q.X. Chen. 2008. A simple disc degeneration model induced by percutaneous needle puncture in the rat tail. Spine 33: 1925–1934.

    PubMed  Google Scholar 

  22. 22.

    Le Maitre, C.L., A. Pockert, D.J. Buttle, A.J. Freemont, and J.A. Hoyland. 2007. Matrix synthesis and degradation in human intervertebral disc degeneration. Biochemical Society Transactions 35: 652–655.

    PubMed  Google Scholar 

  23. 23.

    Sandy, J.D. 2001. Proteoglycan core proteins and catabolic fragments present in tissues and fluids. Methods in molecular biology (Clifton, NJ) 171: 335–345.

    CAS  Google Scholar 

  24. 24.

    Le Maitre, C.L., J.A. Hoyland, and A.J. Freemont. 2007. Catabolic cytokine expression in degenerate and herniated human intervertebral discs: IL-1beta and TNFalpha expression profile. Arthritis Research & Therapy 9: R77.

    Google Scholar 

  25. 25.

    Roelofs, P.D., R.A. Deyo, B.W. Koes, R.J. Scholten, and M.W. van Tulder. 2008. Nonsteroidal anti-inflammatory drugs for low back pain: an updated Cochrane review. Spine 33: 1766–1774.

    PubMed  Google Scholar 

  26. 26.

    Madigan, L., A.R. Vaccaro, L.R. Spector, and R.A. Milam. 2009. Management of symptomatic lumbar degenerative disk disease. The Journal of the American Academy of Orthopaedic Surgeons 17: 102–111.

    PubMed  Google Scholar 

  27. 27.

    Li, K., Y. Li, Z. Ma, and J. Zhao. 2015. Crocin exerts anti-inflammatory and anti-catabolic effects on rat intervertebral discs by suppressing the activation of JNK. International Journal of Molecular Medicine 36: 1291–1299.

    CAS  PubMed  PubMed Central  Google Scholar 

  28. 28.

    Chen, J., J. Xuan, Y.T. Gu, K.S. Shi, J.J. Xie, J.X. Chen, Z.M. Zheng, Y. Chen, X.B. Chen, Y.S. Wu, X.L. Zhang, and X.Y. Wang. 2017. Celastrol reduces IL-1beta induced matrix catabolism, oxidative stress and inflammation in human nucleus pulposus cells and attenuates rat intervertebral disc degeneration in vivo. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 91: 208–219.

    CAS  Google Scholar 

  29. 29.

    Dinarello, C.A. 2011. Interleukin-1 in the pathogenesis and treatment of inflammatory diseases. Blood 117: 3720–3732.

    CAS  PubMed  PubMed Central  Google Scholar 

  30. 30.

    Yang, W., X.H. Yu, C. Wang, W.S. He, S.J. Zhang, Y.G. Yan, J. Zhang, Y.X. Xiang, and W.J. Wang. 2015. Interleukin-1beta in intervertebral disk degeneration. Clinica chimica acta; international journal of clinical chemistry 450: 262–272.

    CAS  PubMed  Google Scholar 

  31. 31.

    Kang, J.D., M. Stefanovic-Racic, L.A. McIntyre, H.I. Georgescu, and C.H. Evans. 1997. Toward a biochemical understanding of human intervertebral disc degeneration and herniation. Contributions of nitric oxide, interleukins, prostaglandin E2, and matrix metalloproteinases. Spine 22: 1065–1073.

    CAS  PubMed  Google Scholar 

  32. 32.

    Sasaki, K., T. Hattori, T. Fujisawa, K. Takahashi, H. Inoue, and M. Takigawa. 1998. Nitric oxide mediates interleukin-1-induced gene expression of matrix metalloproteinases and basic fibroblast growth factor in cultured rabbit articular chondrocytes. Journal of Biochemistry 123: 431–439.

    CAS  PubMed  Google Scholar 

  33. 33.

    Hardy, M.M., K. Seibert, P.T. Manning, M.G. Currie, B.M. Woerner, D. Edwards, A. Koki, and C.S. Tripp. 2002. Cyclooxygenase 2-dependent prostaglandin E2 modulates cartilage proteoglycan degradation in human osteoarthritis explants. Arthritis and Rheumatism 46: 1789–1803.

    CAS  PubMed  Google Scholar 

  34. 34.

    Yoon, S.T., and N.M. Patel. 2006. Molecular therapy of the intervertebral disc. European spine journal : official publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society 15 (Suppl 3): S379–S388.

    Google Scholar 

  35. 35.

    Roberts, S., B. Caterson, J. Menage, E.H. Evans, D.C. Jaffray, and S.M. Eisenstein. 2000. Matrix metalloproteinases and aggrecanase: their role in disorders of the human intervertebral disc. Spine 25: 3005–3013.

    CAS  PubMed  Google Scholar 

  36. 36.

    Bachmeier, B.E., A. Nerlich, N. Mittermaier, C. Weiler, C. Lumenta, K. Wuertz, and N. Boos. 2009. Matrix metalloproteinase expression levels suggest distinct enzyme roles during lumbar disc herniation and degeneration. European spine journal : official publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society 18: 1573–1586.

    Google Scholar 

  37. 37.

    Vo, N.V., R.A. Hartman, T. Yurube, L.J. Jacobs, G.A. Sowa, and J.D. Kang. 2013. Expression and regulation of metalloproteinases and their inhibitors in intervertebral disc aging and degeneration. The spine journal : official journal of the North American Spine Society 13: 331–341.

    Google Scholar 

  38. 38.

    Sun, Z., Z. Yin, C. Liu, H. Liang, M. Jiang, and J. Tian. 2015. IL-1beta promotes ADAMTS enzyme-mediated aggrecan degradation through NF-kappaB in human intervertebral disc. Journal of Orthopaedic Surgery and Research 10: 159.

    PubMed  PubMed Central  Google Scholar 

  39. 39.

    Tu, J., W. Li, Y. Zhang, X. Wu, Y. Song, L. Kang, W. Liu, K. Wang, S. Li, W. Hua, and C. Yang. 2017. Simvastatin inhibits IL-1beta-induced apoptosis and extracellular matrix degradation by suppressing the NF-kB and MAPK pathways in nucleus pulposus cells. Inflammation 40: 725–734.

    CAS  PubMed  Google Scholar 

  40. 40.

    Kong, D., T. Zheng, M. Zhang, D. Wang, S. Du, X. Li, J. Fang, and X. Cao. 2013. Static mechanical stress induces apoptosis in rat endplate chondrocytes through MAPK and mitochondria-dependent caspase activation signaling pathways. PLoS One 8: e69403.

    CAS  PubMed  PubMed Central  Google Scholar 

  41. 41.

    Boutros, T., E. Chevet, and P. Metrakos. 2008. Mitogen-activated protein (MAP) kinase/MAP kinase phosphatase regulation: roles in cell growth, death, and cancer. Pharmacological Reviews 60: 261–310.

    CAS  PubMed  Google Scholar 

  42. 42.

    Seguin, C.A., M. Bojarski, R.M. Pilliar, P.J. Roughley, and R.A. Kandel. 2006. Differential regulation of matrix degrading enzymes in a TNFalpha-induced model of nucleus pulposus tissue degeneration. Matrix biology : journal of the International Society for Matrix Biology 25: 409–418.

    CAS  Google Scholar 

  43. 43.

    Studer, R.K., L.G. Gilbertson, H. Georgescu, G. Sowa, N. Vo, and J.D. Kang. 2008. p38 MAPK inhibition modulates rabbit nucleus pulposus cell response to IL-1. Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society 26: 991–998.

    CAS  Google Scholar 

  44. 44.

    Wako, M., T. Ohba, T. Ando, Y. Arai, K. Koyama, Y. Hamada, A. Nakao, and H. Haro. 2008. Mechanism of signal transduction in tumor necrosis factor-like weak inducer of apoptosis-induced matrix degradation by MMP-3 upregulation in disc tissues. Spine 33: 2489–2494.

    PubMed  Google Scholar 

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Funding

This work was supported by the National Natural Science Foundation of China (81871806 and 81501933), the Zhejiang Public Service Technology Research Program/Social Development (LGF18H060008), the Major Scientific and Technological Project of the Medical and Health Ministry of Zhejiang Province (WKJ-ZJ-1527), the Wenzhou Science and Technology Project (Y20170080), and the Zhejiang Provincial Natural Science Foundation of China (LY17H060008).

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Correspondence to Aimin Wu or Xiangyang Wang.

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The study was performed in accordance with the Declaration of Helsinki and relevant policies in China.

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This study was approved by the Committee of Wenzhou Medical University, and all surgical interventions, treatments, and postoperative animal care procedures were performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals.

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Jin, H., Wang, Q., Wu, J. et al. Baicalein Inhibits the IL-1β-Induced Inflammatory Response in Nucleus Pulposus Cells and Attenuates Disc Degeneration In vivo. Inflammation 42, 1032–1044 (2019). https://doi.org/10.1007/s10753-019-00965-8

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KEY WORDS

  • intervertebral disc degeneration
  • baicalein
  • inflammation
  • NF-κB
  • MAPK