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Indirubin Inhibits LPS-Induced Inflammation via TLR4 Abrogation Mediated by the NF-kB and MAPK Signaling Pathways

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Abstract

Indirubin plays an important role in the treatment of many chronic diseases and exhibits strong anti-inflammatory activity. However, the molecular mode of action during mastitis prophylaxis remains poorly understood. In this study, a lipopolysaccharide (LPS)-induced mastitis mouse model showed that indirubin attenuated histopathological changes in the mammary gland, local tissue necrosis, and neutrophil infiltration. Moreover, indirubin significantly downregulated the production of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α). We explored the mechanism whereby indirubin exerts protective effects against LPS-induced inflammation of mouse mammary epithelial cells (MMECs). The addition of different concentrations of indirubin before exposure of cells to LPS for 1 h significantly attenuated inflammation and reduced the concentrations of the three inflammatory cytokines in a dose-dependent manner. Indirubin downregulated LPS-induced cyclooxygenase-2 (COX-2) and Toll-like receptor 4 (TLR4) expression, inhibited phosphorylation of the LPS-induced nuclear transcription factor-kappa B (NF-kB) P65 protein and its inhibitor IkBα of the NF-kB signaling pathway. Furthermore, indirubin suppressed phosphorylation of P38, extracellular signal-regulated kinase (ERK), and c-Jun NH2-terminal kinase (JNK) of the mitogen-activated protein kinase (MAPK) signal pathways. Thus, indirubin effectively suppressed LPS-induced inflammation via TLR4 abrogation mediated by the NF-kB and MAPK signaling pathways and may be useful for mastitis prophylaxis.

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ACKNOWLEDGMENTS

This project was supported by the Special Fund for China Agriculture Research System (Beef/Yak Cattle) (grant no. CARS-38); the Natural Science Foundation of Hubei Province, China (grant no. 2015CFB435); the National Natural Science Foundation of China (grant no. 31101874); and the Fundamental Research Funds for the Central Universities (grant nos. 2662015PY054 and 2662016PY015).

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  1. The Faculty of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China

    Jin-lun Lai, Yu-hui Liu, Chang Liu, Ming-pu Qi, Rui-ning Liu, Xi-fang Zhu, Qiu-ge Zhou, Ying-yu Chen, Ai-zhen Guo & Chang-min Hu

  2. State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, China

    Ying-yu Chen, Ai-zhen Guo & Chang-min Hu

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  1. Jin-lun Lai
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Lai, Jl., Liu, Yh., Liu, C. et al. Indirubin Inhibits LPS-Induced Inflammation via TLR4 Abrogation Mediated by the NF-kB and MAPK Signaling Pathways. Inflammation 40, 1–12 (2017). https://doi.org/10.1007/s10753-016-0447-7

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