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Forsythin inhibits lipopolysaccharide-induced inflammation by suppressing JAK-STAT and p38 MAPK signalings and ROS production

Inflammation Research volume 63, pages 597–608 (2014)Cite this article

Abstract

Objective

Forsythin (FOR) is an active ingredient extracted from the fruit of the medicinal plant Forsythia suspensa (Thunb.) Vahl. Here, we investigated the effect of FOR on LPS-induced inflammatory response and the underlying molecular mechanisms in RAW264.7 macrophages.

Materials and methods

RAW264.7 cells were pre-treated with or without FOR and then stimulated with or without LPS. The productions of TNF-α, IL-1β, IL-6, PGE2 and NO were determined by ELISA and nitrite analysis, respectively. The expressions of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were measured by Western blotting and RT-PCR analysis. The activations of signaling molecules were detected by Western blotting using phosphorylation specific antibodies. Reactive oxygen species (ROS) production was determined by ROS assay.

Results

LPS-induced productions of IL-1β, IL-6, TNF-α, NO and PGE2 were inhibited by FOR in a dose-dependent manner. FOR also suppressed the LPS-elevated expressions of iNOS and COX-2. Further investigations revealed that FOR significantly inhibited the LPS-induced activations of JAK-STATs and p38 MAPKs, but not of IKKα/β in LPS-stimulated RAW264.7 cells. Additionally, FOR interfered with both JAK-STATs and p38 MAPKs signaling pathways to modulate the expressions of IL-1β, IL-6, TNF-α, iNOS and COX-2. Furthermore, FOR reduced the LPS-induced ROS accumulation, validating that FOR serves as an antioxidant.

Conclusions

Our data suggested that FOR exerts anti-inflammatory action, at least in part, via suppressing LPS-induced activation of JAK-STATs and p38 MAPKs signalings and production of ROS in macrophage cells.

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Acknowledgments

This work was supported by grants from the National Nature Science Foundation of China (Nos. 81072433, 81172798, and 31071000), the Jiangsu Provincial Nature Science Foundation (No. BK2012452), and by the Priority Academic Program Development of Jiangsu Higher Education Institutions (No. 164320H106).

Conflict of interest

The authors declare that they have no conflict of interest.

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Affiliations

  1. Jiangsu Province Key Laboratory for Molecular and Medicine Biotechnology, College of Life Science, Nanjing Normal University, No. 1 Wenyuan Road, Nanjing, 210046, Jiangsu, People’s Republic of China

    Xiaolong Pan, Xiang Cao, Na Li, Yimiao Xu, Qiuyue Wu, Jing Bai, Zhimin Yin & Lei Lan

  2. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, 210093, Jiangsu, People’s Republic of China

    Lan Luo

  3. Collaborative Innovation Center of Biomedicine for Public Hygiene Emergency and Critical Care, Jiangsu Life Sciences and Technology Innovation Park, Nanjing, Jiangsu, People’s Republic of China

    Zhimin Yin, Lan Luo & Lei Lan

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  1. Xiaolong Pan
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Correspondence to Zhimin Yin.

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Responsible Editor: Graham R. Wallace.

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Supplementary material 1 (TIFF 274 kb)

Fig.S1 Cells were pretreated with or without FOR (25, 50, 100, 150, 200 μg/ml) for 2 h. After stimulation with or without LPS (100 ng/ml) for 4 h, the levels of phospho-STAT3 (ser727) and total cellular STAT3 were determined by Western blot analysis.

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Pan, X., Cao, X., Li, N. et al. Forsythin inhibits lipopolysaccharide-induced inflammation by suppressing JAK-STAT and p38 MAPK signalings and ROS production. Inflamm. Res. 63, 597–608 (2014). https://doi.org/10.1007/s00011-014-0731-7

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  • DOI: https://doi.org/10.1007/s00011-014-0731-7

Keywords

  • Forsythin
  • JAK-STATs
  • p38 MAPKs
  • ROS
  • RAW264.7 cells