Skip to main content

Advertisement

SpringerLink
Log in

Divulging the Intricacies of Crosstalk Between NF-kB and Nrf-2/Keap1 Pathway in the Treatment of Arthritis by Dimethyl Fumarate

  • Original Article
  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

The aim of this study was to examine the hypothesis that use of dimethyl fumarate (DMF) may mitigate arthritic symptoms in collagen-induced arthritis (CIA) rats through activation of NF-E2-related factor 2(Nrf-2) and suppression of NF-kB pathway. Arthritis in rats was induced by subcutaneous injection of collagen type II (200 µl) at the base of the tail. After induction arthritic rats were treated with DMF (25 mg/kg b.wt.) for 20 days from the day 25th to 45th. At the end of the study, serum and joint homogenate was used to assess the oxidative stress and cytokines level. In addition, mRNA expression of various genes such as NF-kB, Keap-1 (Kelch-like ECH-associated protein 1) and Nrf-2 was assayed through qRT-PCR in joint tissue. Finally, all these biochemical and molecular results were confirmed by histological and in silico study. Our results showed that decrease in the clinical severity, inflammation, and cell necrosis in DMF-treated rats. This was related with decrease in NF-kB activity and increase in activity of Nrf-2. Treatment with DMF increases the levels of endogenous antioxidant biomarkers glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD) and decreases inflammation. These biochemical and molecular results were further confirmed by performing in silico study that shows DMF strongly inhibits the activation of NF-kB, and conversely at the same time increases the activity of Nrf-2 that means a significantly lower amount of inflammatory mediators and oxidants was produced. Decrease in inflammation leads to preserving the joint architecture and alleviation from clinical symptoms of arthritis. Collectively, these results indicate that Nrf-2 activation protects against arthritic symptoms.

Graphical Abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

Data Availability

The data used to support the findings of this study are available from the corresponding author upon request.

Abbreviations

CIA:

Collagen-induced arthritis

LPO:

Lipid Peroxidation

GSH:

Reduced glutathione

CAT:

Catalase

SOD:

Super Oxide Dismutase

TNF-α:

Tumor necrosis factor-α

IL-1β:

Interleukin-1β

IL-10:

Interleukin-10

COX-2:

Cyclooxygenase-2

NF-kBp65:

Nuclear factor-kBp65

Nrf-2:

Nuclear factor erythroid 2–related factor 2

MDA:

Malondialdehyde

References

  1. McInnes, I. B., & Schett, G. (2007). Cytokines in the pathogenesis of rheumatoid arthritis. Nature reviews. Immunology, 7(6), 429–442.

    Article  CAS  PubMed  Google Scholar 

  2. Müller-Ladner, U., Ospelt, C., Gay, S., Distler, O., & Pap, T. (2007). Cells of the synovium in rheumatoid arthritis. Synovial fibroblasts. Arthritis research & therapy, 9(6), 223.

    Article  Google Scholar 

  3. Jones, B. A., Riegsecker, S., Rahman, A., Beamer, M., Aboualaiwi, W., Khuder, S. A., & Ahmed, S. (2013). Role of ADAM-17, p38 MAPK, cathepsins, and the proteasome pathway in the synthesis and shedding of fractalkine/CX3 CL1 in rheumatoid arthritis. Arthritis and rheumatism, 65(11), 2814–2825.

    Article  CAS  PubMed  Google Scholar 

  4. Iwamoto, T., Okamoto, H., Toyama, Y., & Momohara, S. (2008). Molecular aspects of rheumatoid arthritis: Chemokines in the joints of patients. The FEBS journal, 275(18), 4448–4455.

    Article  CAS  PubMed  Google Scholar 

  5. Lawrence, T. (2009). The nuclear factor NF-kappaB pathway in inflammation. Cold Spring Harbor Perspectives in Biology, 1(6), a001651.

    Article  PubMed Central  PubMed  Google Scholar 

  6. Liu, T., Zhang, L., Joo, D., & Sun, S. C. (2017). NF-κB signaling in inflammation. Signal Transduction And Targeted Therapy, 2, 17023.

    Article  PubMed Central  PubMed  Google Scholar 

  7. Traenckner, E. B., Wilk, S., & Baeuerle, P. A. (1994). A proteasome inhibitor prevents activation of NF-kappa B and stabilizes a newly phosphorylated form of I kappa B-alpha that is still bound to NF-kappa B. The EMBO Journal, 13(22), 5433–5441.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Janssen-Heininger, Y. M., Poynter, M. E., & Baeuerle, P. A. (2000). Recent advances towards understanding redox mechanisms in the activation of nuclear factor kappaB. Free Radical Biology & Medicine, 28(9), 1317–1327.

    Article  CAS  Google Scholar 

  9. Kavian, N., Mehlal, S., Jeljeli, M., Saidu, N., Nicco, C., Cerles, O., Chouzenoux, S., Cauvet, A., Camus, C., Ait-Djoudi, M., Chéreau, C., Kerdine-Römer, S., Allanore, Y., & Batteux, F. (2018). The Nrf2-antioxidant response element signaling pathway controls fibrosis and autoimmunity in scleroderma. Frontiers In Immunology, 9, 1896.

    Article  PubMed Central  PubMed  Google Scholar 

  10. Payne, R. (2000). Limitations of NSAIDs for pain management: Toxicity or lack of efficacy? The Journal Of Pain, 1(3 Suppl), 14–18.

    Article  CAS  PubMed  Google Scholar 

  11. Papadopoulou, A., D’Souza, M., Kappos, L., & Yaldizli, O. (2010). Dimethyl fumarate for multiple sclerosis. Expert Opinion On Investigational Drugs, 19(12), 1603–1612.

    Article  CAS  PubMed  Google Scholar 

  12. Lee, D. H., Stangel, M., Gold, R., & Linker, R. A. (2013). The fumaric acid ester BG-12: A new option in MS therapy. Expert Review Of Neurotherapeutics, 13(8), 951–958.

    Article  CAS  PubMed  Google Scholar 

  13. Kobayashi, K., Tomiki, H., Inaba, Y., Ichikawa, M., Kim, B. S., & Koh, C. S. (2015). Dimethyl fumarate suppresses Theiler’s murine encephalomyelitis virus-induced demyelinating disease by modifying the Nrf2-Keap1 pathway. International Immunology, 27(7), 333–344.

    Article  CAS  PubMed  Google Scholar 

  14. Bomprezzi, R. (2015). Dimethyl fumarate in the treatment of relapsing-remitting multiple sclerosis: An overview. Therapeutic Advances In Neurological Disorders, 8(1), 20–30.

    Article  PubMed Central  PubMed  Google Scholar 

  15. Campo, G. M., Avenoso, A., Campo, S., Ferlazzo, A. M., Altavilla, D., & Calatroni, A. (2003). Efficacy of treatment with glycosaminoglycans on experimental collagen-induced arthritis in rats. Arthritis Research & Therapy, 5(3), R122–R131.

    Article  CAS  Google Scholar 

  16. Holmdahl, R., Carlsen, S., Mikulowska, A., Vestberg, M., Brunsberg, U., Hansson, A., Sunduall, M., Jans-son, L., & Pettersson, U. (1998). Genetic analysis of murine models for rheumatoid arthritis. In K. W. Adolpho (Ed.), Human Genome Methods (pp. 215–238). CRC Press.

    Google Scholar 

  17. Robles, L., Vaziri, N. D., Li, S., Takasu, C., Masuda, Y., Vo, K., Farzaneh, S. H., Stamos, M. J., & Ichii, H. (2015). Dimethyl fumarate ameliorates acute pancreatitis in rodent. Pancreas, 44(3), 441–447.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Satoh, K. (1978). Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clinica Chimica Acta, 90(1), 37–43.

    Article  CAS  Google Scholar 

  19. Tietze, F. (1969). Enzymic method for quantitative deter-mination of nanogram amounts of total and oxi-dized glutathione: Applications to mammalian blood and other tissues. Analytical Biochemistry, 27(3), 502–522.

    Article  CAS  PubMed  Google Scholar 

  20. Hyc, A., Osiecka-Iwan, A., Dziunycz, P., & Moskalewski, S. (2007). Preparation of rat synovial membrane for studies of cytokine secretion. Folia Histochemica Et Cytobiologica, 45(1), 57–60.

    CAS  PubMed  Google Scholar 

  21. Schmittgen, T. D., Zakrajsek, B. A., Mills, A. G., Gorn, V., Singer, M. J., & Reed, M. W. (2000). Quantitative reverse transcription-polymerase chain reaction to study mRNA decay: Comparison of endpoint and real-time methods. Analytical biochemistry, 285(2), 194–204.

    Article  CAS  PubMed  Google Scholar 

  22. Morris, G. M., Goodsell, D. S., Halliday, R. S., Huey, R., Hart, W. E., Belew, R. K., & Olson, A. J. (1998). Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function. Journal Of Computational Chemistry, 19(14), 1639–1662.

    Article  CAS  Google Scholar 

  23. Solis, F. J., & Wets, R. J. (1921). Minimization by random search techniques. Mathematics Of Operations Research, 6(1), 19–30.

    Article  Google Scholar 

  24. Khan, M. A., Rabbani, G., Kumari, M., et al. (2022). Ellagic acid protects type II collagen induced arthritis in rat via diminution of IKB phosphorylation and suppression IKB-NF-kB complex activation: In vivo and in silico study. Inflammopharmacol, 30, 1729–1743.

    Article  CAS  Google Scholar 

  25. Stuart, J. M., Townes, A. S., & Kang, A. H. (1982). The role of collagen autoimmunity in animal models and human diseases. The Journal of Investigative Dermatology, 79(Suppl 1), 121s–127s.

    Article  PubMed  Google Scholar 

  26. Wooley, P. H., Luthra, H. S., Krco, C. J., Stuart, J. M., & David, C. S. (1984). Type II collagen-induced arthritis in mice. II. Passive transfer and suppression by intravenous injection of anti-type II collagen antibody or free native type II collagen. Arthritis and Rheumatism, 27(9), 1010–1017.

    Article  CAS  PubMed  Google Scholar 

  27. Taysi, S., Polat, F., Gul, M., Sari, R. A., & Bakan, E. (2001). Lipid peroxidation, some extracellular anti-oxidants, and anti-oxidant enzymes in serum of patients with rheumatoid arthritis. Rheumatology International, 21(5), 200–4.

    Article  Google Scholar 

  28. Khan, M. A., Subramaneyaan, M., Arora, V. K., Banerjee, B. D., & Ahmed, R. S. (2015). Effect of Withania somnifera (Ashwagandha) root extract on amelioration of oxidative stress and autoantibodies production in collagen-induced arthritic rats. Journal of Complementary and Integrative Medicine, 12(2), 117–125.

    Article  PubMed  Google Scholar 

  29. Jambale, T. A., Halyal, S. S., & Jayaprakash Murthy, D. S. (2014). Study of serum malondialdehyde, nitric oxide, vitamin E levels in patients with rheumatoid arthritis. International Journal of Medical Research and Health Sciences, 3(2), 322–325.

    Article  Google Scholar 

  30. Bharath, S., Hsu, M., Kaur, D., Rajagopalan, S., & Andersen, J. K. (2002). Glutathione, iron and Parkinson’s disease. Biochemical pharmacology, 64(5–6), 1037–1048.

    Article  CAS  PubMed  Google Scholar 

  31. Freeman, B. A., & Crapo, J. D. (1982). Biology of disease: Free radicals and tissue injury. Laboratory investigation; a journal of technical methods and pathology, 47(5), 412–426. [32]. Niture, S. K., Khatri, R., & Jaiswal, A. K. (2014). Regulation of Nrf2-an update. Free Radical Biology & Medicine, 66, 36–44.

    Google Scholar 

  32. Linker, R. A., Lee, D. H., Ryan, S., van Dam, A. M., Conrad, R., Bista, P., Zeng, W., Hronowsky, X., Buko, A., Chollate, S., Ellrichmann, G., Brück, W., Dawson, K., Goelz, S., Wiese, S., Scannevin, R. H., Lukashev, M., & Gold, R. (2011). Fumaric acid esters exert neuroprotective effects in neuroinflammation via activation of the Nrf2 antioxidant pathway. Brain : A Journal of Neurology, 134(Pt 3), 678–692.

    Article  PubMed  Google Scholar 

  33. Scannevin, R. H., Chollate, S., Jung, M. Y., Shackett, M., Patel, H., Bista, P., Zeng, W., Ryan, S., Yamamoto, M., Lukashev, M., & Rhodes, K. J. (2012). Fumarates promote cytoprotection of central nervous system cells against oxidative stress via the nuclear factor (erythroid-derived 2)-like 2 pathway. The Journal of Pharmacology and Experimental Therapeutics, 341(1), 274–284.

    Article  CAS  PubMed  Google Scholar 

  34. Yang, Y., Jiang, S., Yan, J., Li, Y., Xin, Z., Lin, Y., & Qu, Y. (2015). An overview of the molecular mechanisms and novel roles of Nrf2 in neurodegenerative disorders. Cytokine & Growth Factor Reviews, 26(1), 47–57.

    Article  CAS  Google Scholar 

  35. Reddy, N. M., Kleeberger, S. R., Kensler, T. W., Yamamoto, M., Hassoun, P. M., & Reddy, S. P. (2009). Disruption of Nrf2 impairs the resolution of hyperoxia-induced acute lung injury and inflammation in mice. Journal of immunology (Baltimore, Md. : 1950), 182(11), 7264–7271.

    Article  CAS  PubMed  Google Scholar 

  36. Wang, H., Khor, T. O., Saw, C. L., Lin, W., Wu, T., Huang, Y., & Kong, A. N. (2010). Role of Nrf2 in suppressing LPS-induced inflammation in mouse peritoneal macrophages by polyunsaturated fatty acids docosahexaenoic acid and eicosapentaenoic acid. Molecular pharmaceutics, 7(6), 2185–2193.

    Article  CAS  PubMed  Google Scholar 

  37. Ganesh Yerra, V., Negi, G., Sharma, S. S., & Kumar, A. (2013). Potential therapeutic effects of the simultaneous targeting of the Nrf2 and NF-κB pathways in diabetic neuropathy. Redox Biology, 1(1), 394–397.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  38. Isomäki, P., & Punnonen, J. (1997). Pro- and anti-inflammatory cytokines in rheumatoid arthritis. Annals of Medicine, 29(6), 499–507.

    Article  PubMed  Google Scholar 

  39. Szekanecz, Z., Halloran, M. M., Volin, M. V., Woods, J. M., Strieter, R. M., Kenneth Haines, G., Kunkel, S. L., 3rd., Burdick, M. D., & Koch, A. E. (2000). Temporal expression of inflammatory cytokines and chemokines in rat adjuvant-induced arthritis. Arthritis and Rheumatism, 43(6), 1266–1277.

    Article  CAS  PubMed  Google Scholar 

  40. Wilms, H., Sievers, J., Rickert, U., Rostami-Yazdi, M., Mrowietz, U., & Lucius, R. (2010). Dimethylfumarate inhibits microglial and astrocytic inflammation by suppressing the synthesis of nitric oxide, IL-1beta, TNF-alpha and IL-6 in an in-vitro model of brain inflammation. Journal of Neuroinflammation, 7, 30.

    Article  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgements

We acknowledge the Department of Biochemistry, University College of Medical Sciences & GTB Hospital, New Delhi, India, for proving us basic infrastructure for performing experiments.

Author information

Authors and Affiliations

  1. Department of Biochemistry, University College of Medical Sciences & GTB Hospital, Dilshad Garden, Delhi, 110095, India

    Mahmood Ahmad Khan & Rafat Sultana Ahmed

  2. Nano Diagnostics & Devices (NDD), IT Medical Fusion Center, 350-27 Gumidae-ro, Gumi Si, Gyeongbuk, 39253, Republic of Korea

    Gulam Rabbani

  3. Department of Biochemistry, Santosh Deemed to Be University, Ghaziabad, U.P, India

    Juhi Aggarawal

Authors
  1. Mahmood Ahmad Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gulam Rabbani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Juhi Aggarawal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rafat Sultana Ahmed
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

MAK, RSA: designing of work, performing experimental work, data collection, and analysis; JA, GR performed the in silico study. All the authors read and approved the final manuscript.

Corresponding author

Correspondence to Mahmood Ahmad Khan.

Ethics declarations

Ethical Approval

The experimental protocol was approved by the Institutional Animal Ethics Committee.

Consent of Participate

Not applicable.

Competing Interests

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Highlights

This is first time we have reported the anti-arthritic effect of DMF through cross talk between NF-kB and Nrf-2/Keap-1 pathway. In our study, we found that DMF preserved the clinical severity in CIA rats through activation of Nrf-2 and simultaneous inhibition of NF-kB. Furthermore, in silico study was also performed to correlate the above results, and we found that DMF inhibits the activation of NF-kB. These findings may become beneficial, to develop a new therapeutic strategy in the treatment of RA.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Khan, M.A., Rabbani, G., Aggarawal, J. et al. Divulging the Intricacies of Crosstalk Between NF-kB and Nrf-2/Keap1 Pathway in the Treatment of Arthritis by Dimethyl Fumarate. Appl Biochem Biotechnol 195, 4177–4195 (2023). https://doi.org/10.1007/s12010-023-04324-0

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-023-04324-0

Keywords

Search

Navigation