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Daphnetin reduces endotoxin lethality in mice and decreases LPS-induced inflammation in Raw264.7 cells via suppressing JAK/STATs activation and ROS production

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Abstract

Objective

Here, we used various approaches to investigate the suppressive role of daphnetin in LPS-induced inflammatory response, with the goal to understand the underlining molecular mechanism by which daphnetin regulated these processes.

Methods

We examined the survival rate and the lung injury in the mice model of LPS-induced endotoxemia. The production of pro-inflammatory factors including tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), IL-6, nitric oxide (NO), and prostaglandin E2 (PGE2) was measured by ELISA and nitrite analysis, respectively. The expression of inducible NO synthase (iNOS), cyclooxygenase 2 (COX-2), and the activation of signaling molecules was determined by immunoblotting. The production of reactive oxygen species (ROS) was measured by the ROS assay.

Results

In vivo study showed that daphnetin enhanced the survival rate and reduced the lung injury in mice with LPS-induced endotoxemia. Both in vivo and in vitro study showed that daphnetin prevented the production of pro-inflammatory factors including TNF-α, IL-1β, IL-6, NO, and PGE2 after LPS challenge. In Raw264.7 cells, we found that daphnetin reduced LPS-induced expression of iNOS and COX-2, and suppressed LPS-induced ROS production. In addition, we found that daphnetin suppressed the activation of JAK/STATs pathway and inhibited the nucleus import of STAT1 and STAT3.

Conclusions

Here, our results indicate that daphnetin shows anti-inflammatory properties, at least in part, through suppressing LPS-induced activation of JAK/STATs cascades and ROS production.

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Abbreviations

DFN:

Daphnetin

LPS:

Lipopolysaccharide

IL:

Interleukin

TNF:

Tumor necrosis factor

NO:

Nitric oxide

PGE2:

Prostaglandin E2

iNOS:

Inducible NO synthase

COX-2:

Cyclooxygenase 2

JAK:

Janus protein tyrosine kinase

STAT:

Signal transducers and activators of transcription

MAPKs:

Mitogen-activated protein kinases

ROS:

Reactive oxygen species

I.P. injection:

Intraperitoneal injection

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Acknowledgements

This work was supported by Grants from the National Natural Science Foundation of China (Nos. 81471557, 31571166, 81671565, and 31500628) a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (No. 164320H106) and Jiangsu Major Nature Science Foundation of High Education (No. 12KJA180006). We thank lab members for their encouragements.

Author contributions

Conceived and designed the experiments: ZMY LS. Performed the experiments: LS. Analysis the data: ZMY L. Lan LS L. Luo TZ JW XMS. Contributed reagents/materials/analysis tools: ZMY L. Lan L. Luo LS. Wrote the paper: LS ZMY L. Luo.

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Authors and Affiliations

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

    Lei Shen, Ting Zhou, Jing Wang, Xiumei Sang, Lei Lan & Zhimin Yin

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

    Lan Luo

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  1. Lei Shen
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  2. Ting Zhou
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  3. Jing Wang
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  5. Lei Lan
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  6. Lan Luo
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Correspondence to Lei Lan, Lan Luo or Zhimin Yin.

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Responsible Editor: Artur Bauhofer.

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Shen, L., Zhou, T., Wang, J. et al. Daphnetin reduces endotoxin lethality in mice and decreases LPS-induced inflammation in Raw264.7 cells via suppressing JAK/STATs activation and ROS production. Inflamm. Res. 66, 579–589 (2017). https://doi.org/10.1007/s00011-017-1039-1

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  • DOI: https://doi.org/10.1007/s00011-017-1039-1

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