Journal of Natural Medicines

, Volume 71, Issue 2, pp 349–356 | Cite as

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

  • Wen-Xiao Sun
  • Yan Liu
  • Wei Zhou
  • He-Wei Li
  • Jian Yang
  • Zhen-Bing ChenEmail author
Original Paper


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.


Rheumatoid arthritis Inflammation Lithospermum erythrorhizon Protein kinase C Protein synthesis mRNA stability 



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

Compliance with ethical standards

Conflict of interest

The authors declare that no conflict of interests exists.

Supplementary material

11418_2016_1064_MOESM1_ESM.tif (556 kb)
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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Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan 2016

Authors and Affiliations

  • Wen-Xiao Sun
    • 1
  • Yan Liu
    • 1
  • Wei Zhou
    • 1
  • He-Wei Li
    • 1
  • Jian Yang
    • 1
  • Zhen-Bing Chen
    • 2
    Email author
  1. 1.Department of Orthopedic and Hand Surgery, Liyuan HospitalHuazhong University of Science and TechnologyWuhanChina
  2. 2.Department of Orthopedic and Hand Surgery, Union Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina

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