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Involvement of Na+, K+-ATPase and its inhibitors in HuR-mediated cytokine mRNA stabilization in lung epithelial cells

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Abstract

Increasing evidence demonstrates that Na+, K+-ATPase plays an important role in pulmonary inflammation, but the mechanism remains largely unknown. In this study, we used cardiotonic steroids as Na+, K+-ATPase inhibitors to explore the possible involvement of Na+, K+-ATPase in pulmonary epithelial inflammation. The results demonstrated that mice after ouabain inhalation developed cyclooxygenase-2-dependent acute lung inflammation. The in vitro experiments further confirmed that Na+, K+-ATPase inhibitors significantly stimulated cyclooxygenase-2 expression in lung epithelial cells of human or murine origin, the process of which was participated by multiple cis-elements and trans-acting factors. Most importantly, we first described here that Na+, K+-ATPase inhibitors could evoke a significant Hu antigen R nuclear export in lung epithelial cells, which stabilized cyclooxygenase-2 mRNA by binding with a proximal AU-rich element within its 3′-untranslated region. In conclusion, HuR-mediated mRNA stabilization opens new avenues in understanding the importance of Na+, K+-ATPase, as well as its inhibitors in inflammation.

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Abbreviations

Act D:

Actinomycin D

ALI:

Acute lung injury

ARDS:

Acute respiratory distress syndrome

ARE:

AU-rich elements

BALF:

Bronchoalveolar lavage fluid

COX-2:

Cyclooxygenase-2

CTS:

Cardiotonic steroids

DMEM:

Dulbecco’s modified Eagle’s medium

DN:

Dominant negative

EO:

Endogenous ouabain

ERK:

Extracellular regulated protein kinases

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HA:

Hemagglutinin

H&E:

Hematoxylin and eosin

HRP:

Horseradish peroxidase

HuR:

Hu antigen R

IκB:

Inhibitory subunit of nuclear factor kappa B

LPS:

Lipopolysaccharide

MAPK:

Mitogen-activated protein kinase

MK2:

MAPK-activated protein kinase 2

MPO:

Myeloperoxidase

NO:

Nitric oxide

PGE2 :

Prostaglandin E2

ROS:

Reactive oxygen species

TNF:

Tumor necrosis factor

UTR:

Untranslated region

VEGF:

Vascular endothelial growth factor

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Acknowledgments

This work was supported by the Natural Science Fund of China (30470644, 30600502, 30973528 to Y. W., 30425009, 30330530 to H. Z.C., 30730107 to X. Q). Natural Science Fund of Jiangsu Province (BK2008272 to Y. W.). Natural Science Fund of Shanghai (10ZR1438400 to B.J.J.). Open project of Jiangsu Key Lab for Drug Efficiency and Safety Evaluation (P09008); and the Ministry of Education of China (TRAPOY 199028418).

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Authors and Affiliations

  1. The State Key Lab of Pharmaceutical Biotechnology, College of Life Sciences, Nanjing University, Nanjing, 210093, China

    Su Feng, Wei Chen, Dan Cao, Fang-Yuan Gong, Wei Cheng, Shun Cheng, Qiang Xu, Zi-Chun Hua & Wu Yin

  2. Department of Anesthesia and Intensive Care Unit, Changhai Hospital, Affiliated Hospital of the Second Military Medical University, Shanghai, 200433, China

    Jinjun Bian

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  1. Su Feng
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  2. Wei Chen
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  3. Dan Cao
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  4. Jinjun Bian
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  7. Shun Cheng
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  9. Zi-Chun Hua
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Correspondence to Zi-Chun Hua or Wu Yin.

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S. Feng and W. Chen contributed equally to this paper.

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Feng, S., Chen, W., Cao, D. et al. Involvement of Na+, K+-ATPase and its inhibitors in HuR-mediated cytokine mRNA stabilization in lung epithelial cells. Cell. Mol. Life Sci. 68, 109–124 (2011). https://doi.org/10.1007/s00018-010-0444-1

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