Abstract
We investigated the efficacy of the traditional herbal extract 6-bromoindirubin-3′-oxime (BIO) against lipopolysaccharide (LPS)-induced mastitis in mice and inflammatory signaling in mouse mammary epithelial cells (MMECs). In vivo, breast inflammation scores and enzyme-linked immunosorbent assay (ELISA) detection of pro-inflammatory factor expression were used to assess the effect of BIO against mastitis. In vitro, the effects of BIO on LPS-induced changes in the expression levels of pro-inflammatory factors, anti-inflammatory cytokines, and signaling factors of the toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) and TLR4/mitogen-activated protein kinase (MAPK) pathways were examined by qRT-PCR and ELISA. In LPS-injected mice, BIO pretreatment downregulated the expression of the pro-inflammatory factors interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and myeloperoxidase (MPO) in mammary glands and reduced inflammatory lesions in breast tissue. In MMECs, BIO pretreatment downregulated the LPS-induced expression of IL-1β, IL-6, and TNF-α. Further, BIO inhibited both the expression and phosphorylation of TLR4/NF-κB and TLR4/MAPK signaling factors. Thus, BIO downregulates IL-6, IL-1β, TNF-α, and MPO expression, upregulates IL-10 expression, and suppresses LPS-induced inflammation by inhibiting the TLR4/NF-κB and TLR4/MAPK pathways. BIO may be a potential treatment agent for mastitis and other inflammatory diseases.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ouyang, W., S. Rutz, N.K. Crellin, P.A. Valdez, and S.G. Hymowitz. 2011. Regulation and functions of the IL-10 family of cytokines in inflammation and disease. Annual Review of Immunology 29 (29): 71–109.
Kiku, Y., T.O. Zawa, H.T. Akahashi, et al. 2017. Effect of intramammary infusion of recombinant bovine GM-CSF and IL-8 on CMT score, somatic cell count, and milk mononuclear cell populations in Holstein cows with Staphylococcus aureus subclinical mastitis. Veterinary Research Communications: 1–8.
Gilbert, F.B., P.C. Unha, K.J. Ensen, et al. 2013. Differential response of bovine mammary epithelial cells to Staphylococcus aureus or Escherichia coli agonists of the innate immune system. Veterinary Research 44 (1): 40.
Wang, C., X. Bai, and C. Wang. 2014. Traditional Chinese medicine: a treasured natural resource of anticancer drug research and development. American Journal of Chinese Medicine 42 (3): 1450035.
Lai, J.L., Y.H. Liu, C. Liu, M.P. Qi, R.N. Liu, X.F. Zhu, Q.G. Zhou, Y.Y. Chen, A.Z. Guo, and C.M. Hu. 2017. Indirubin inhibits LPS-induced inflammation via TLR4 abrogation mediated by the NF-κB and MAPK signaling pathways. Inflammation. 40 (1): 1–12.
Engel, M.A., C.A. Kellermann, G. Burnat, E.G. Hahn, T. Rau, and P.C. Konturek. 2010. Mice lacking cannabinoid CB1-, CB2-receptors or both receptors show increased susceptibility to trinitrobenzene sulfonic acid (TNBS)-induced colitis. Journal of Physiology and Pharmacology 61 (1): 89–97.
Lai, J.L., Y.H. Liu, Y.C. Peng, P. Ge, C.F. He, C. Liu, Y.Y. Chen, A.Z. Guo, and C.M. Hu. 2017. Indirubin treatment of lipopolysaccharide-induced mastitis in a mouse model and activity in mouse mammary epithelial cells. Mediators of Inflammation: 3082805. https://doi.org/10.1155/2017/3082805.
Michlewska, S.M.C.F., I.I. Dransfield, and A. Rossi. 2009. Macrophage phagocytosis of apoptotic neutrophils is critically regulated by the opposing actions of pro-inflammatory and anti-inflammatory agents: key role for TNF-alpha. FASEB Journal 23 (3): 844–854.
Lv, X., K. Fu, W. Li, Y. Wang, J. Wang, H. Li, W. Tian, and R. Cao. 2015. TIIA attenuates LPS-induced mouse endometritis by suppressing the NF-κB signaling pathway. Canadian Journal of Physiology and Pharmacology 93 (11): 967–971.
Fang, W.F., Y.M. Chen, C.Y. Lin, H.L. Huang, H. Yeh, Y.T. Chang, K.T. Huang, and M.C. Lin. 2018. Histone deacetylase 2 (HDAC2) attenuates lipopolysaccharide (LPS)-induced inflammation by regulating PAI-1 expression. Journal of Inflammation 15 (1): 3.
Braig, S., C.A. Kressirer, J. Liebl, F. Bischoff, S. Zahler, L. Meijer, and A.M. Vollmar. 2013. Indirubin derivative 6 BIO suppresses metastasis. Cancer Research 73 (19): 6004–6012.
Eljaafari, A., M. Robert, M. Chehimi, S. Chanon, C. Durand, G. Vial, N. Bendridi, A.M. Madec, E. Disse, M. Laville, J. Rieusset, E. Lefai, H. Vidal, and L. Pirola. 2015. Adipose tissue-derived stem cells from obese subjects contribute to inflammation and reduced insulin response in adipocytes through differential regulation of the Th1/Th17 balance and monocyte activation. Diabetes 64 (7): 2477–2488.
Banks, W.A., A.J. Kastin, and E.G. Gutierrez. 1994. Penetration of interleukin-6 across the murine blood-brain barrier. Neuroscience Letters 179 (1–2): 53–56.
Schaper, F., and S. Rose-John. 2015. Interleukin-6: Biology, signaling and strategies of blockade. Cytokine & Growth Factor Reviews 26 (5): 475–487.
Kearney, C.J., S.P. Cullen, G.A. Tynan, C.M. Henry, D. Clancy, E.C. Lavelle, and S.J. Martin. 2015. Necroptosis suppresses inflammation via termination of TNF- or LPS-induced cytokine and chemokine production. Cell Death and Differentiation 22 (8): 1313–1327.
Siqueira, M.B., A. Kroner, E.I. Girolami, et al. 2015. Role of IL-10 in resolution of inflammation and functional recovery after peripheral nerve injury. The Journal of Neuroscience: the Official Journal of the Society for Neuroscience 35 (50): 16431–16442.
Jr, J.C., and R. Medzhitov. 2002. Innate immune recognition. Annual Review of Immunology 20 (1): 197–216.
Aratani, Y. 2018. Myeloperoxidase: its role for host defense, inflammation, and neutrophil function. Archives of Biochemistry and Biophysics 640: 47–52.
Zhang, X., Y. Wang, C. Xiao, Z. Wei, J. Wang, Z. Yang, and Y. Fu. 2017. Resveratrol inhibits LPS-induced mice mastitis through attenuating the MAPK and NF-κB signaling pathway. Microbial Pathogenesis 107: 462–467.
May, M.J., and S. Ghosh. 1998. NF-kappaB and Rel proteins: evolutionarily conserved mediators of immune responses. Immunology Today 19.
Serasanambati, M., and S.R. Chilakapati. 2016. Function of nuclear factor kappa B (NF-κB) in human diseases—a review. South Indian Journal of Biological Sciences 2 (4): 368–387.
Liu, Y., H. Yin, M. Zhao, and Q. Lu. 2014. TLR2 and TLR4 in autoimmune diseases: a comprehensive review. Clinical Reviews in Allergy & Immunology 47 (2): 136–147.
Thalhamer, T., M.A. Mcgrath, and M.M. Harnett. 2008. MAPKs and their relevance to arthritis and inflammation. Rheumatology 47 (4): 409–414.
Kyriakis, J.M., and J. Avruch. 2012. Mammalian MAPK signal transduction pathways activated by stress and inflammation: a 10-year update. Physiological Reviews 92 (2): 689–737.
Acknowledgments
We would like to thank all our coworkers in the State Key Laboratory of Agricultural Microbiology (Wuhan, China) for useful discussions. This work was supported by the National Key Research and Development Plan (No. 2016YFD0500906), and the special fund for the China Agriculture Research System (Beef/Yak cattle) (No. CARS-38), and the Research Fund for National Distinguished Scholars in Agriculture Research and Technical Innovative Team.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
All co-authors implicated in this research approved this article to be published. The authors declare that they have no conflict of interest.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Liu, C., Tang, X., Zhang, W. et al. 6-Bromoindirubin-3′-Oxime Suppresses LPS-Induced Inflammation via Inhibition of the TLR4/NF-κB and TLR4/MAPK Signaling Pathways. Inflammation 42, 2192–2204 (2019). https://doi.org/10.1007/s10753-019-01083-1
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10753-019-01083-1