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Shikonin inhibits TNF-α production through suppressing PKC-NF-κB-dependent decrease of IL-10 in rheumatoid arthritis-like cell model

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Abstract

Shikonin, a major effective component in the Chinese herbal medicine Lithospermum erythrorhizon Sieb., exhibits an anti-inflammatory property towards rheumatoid arthritis (RA), but the potential mechanism is unclear. Our aim was to investigate the mechanism of shikonin on the lipopolysaccharide (LPS)-induced fibroblast-like synoviocyte (LiFLS) inflammation model. Fibroblast-like synoviocytes (FLSs) were treated with 200 μg/ml of LPS for 24 h to establish the RA-like model, LiFLS. FLSs were pretreated with shikonin (0.1–1 μM) for 30 min in the treatment groups. Quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assays were used to detect mRNA and protein levels of interleukin (IL)-10 and tumor necrosis factor (TNF)-α. Signal proteins involved in IL-10 production were analyzed by Western blotting. Shikonin significantly reversed the inhibitory effects of LPS on IL-10 expression in FLSs by inactivating the PKC-NF-κB pathway. In addition, shikonin inhibited LPS-induced TNF-α expression in FLSs, and this effect was markedly diminished by IL-10-neutralizing antibody. The IL-10-mediated suppression of TNF-α transcription was demonstrated by no response to the protein synthesis inhibitor cyclohexamide and no mRNA decay. Shikonin inhibits LPS-induced TNF-α production in FLSs through suppressing the PKC-NF-κB-dependent decrease in IL-10, and this study also highlights the potential application of shikonin in the treatment of RA.

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Acknowledgements

This work was supported by grants from the Science and Technology Planning Natural Scientific Foundation of Hubei Province of China (2010CDB07501).

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    Authors and Affiliations

    1. Department of Orthopedic and Hand Surgery, Liyuan Hospital, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, 430077, Hubei, China

      Wen-Xiao Sun, Yan Liu, Wei Zhou, He-Wei Li & Jian Yang

    2. Department of Orthopedic and Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, Hubei, China

      Zhen-Bing Chen

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    1. Wen-Xiao Sun
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    Correspondence to Zhen-Bing Chen.

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    W.-X. Sun and Y. Liu contributed equally to this work.

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    11418_2016_1064_MOESM1_ESM.tif

    Supplementary Fig. 1. Identification of FLSs by microscope and flow cytometry. LiFLSs was identified by (a) microscopy (200× and 400×) or (b) flow cytometry using antibodies anti-vimentin and anti-CD68. (TIFF 556 kb)

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    Sun, WX., Liu, Y., Zhou, W. et al. Shikonin inhibits TNF-α production through suppressing PKC-NF-κB-dependent decrease of IL-10 in rheumatoid arthritis-like cell model. J Nat Med 71, 349–356 (2017). https://doi.org/10.1007/s11418-016-1064-3

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