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Reactive oxygen species mediate Jak2/Stat3 activation and IL-8 expression in pulmonary epithelial cells stimulated with lipid-associated membrane proteins from Mycoplasma pneumoniae

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Abstract

Objective

To investigate the involvement of reactive oxygen species (ROS) in the activation of Janus kinase2 (Jak2)/signal transducers and activators of transcription3 (Stat3), and IL-8 expression in pulmonary epithelial cells stimulated with lipid-associated membrane proteins (LAMP) from Mycoplasma pneumoniae using a known antioxidant, N-acetylcysteine (NAC).

Methods

Pulmonary epithelial A549 cells were treated with or without NAC in the presence or absence of LAMP. Intracellular ROS levels were detected by fluorescent analysis for fluorescent dichlorofluorescein. mRNA expression of IL-8 was analyzed by reverse transcription-polymerase chain reaction. IL-8 protein in the medium was determined by enzyme-linked immunosorbent assay. Activation of Jak2/Stat3 was determined by the increases in phospho-specific forms of Jak2/Stat3 compared to total forms of Jak2/Stat3 by western blotting. Stat3–DNA binding activity was assessed by electrophoretic mobility shift assay.

Results

LAMP increased the level of ROS, phosphorylation of Jak2/Stat3, Stat3–DNA binding activity, and IL-8 expression in A549 cells, which were inhibited by NAC dose-dependently.

Conclusion

LAMP of M. pneumoniae induces the production of ROS, Jak2/Stat3 activation, and IL-8 induction in A549 cells. Antioxidants such as NAC may be beneficial for preventing pulmonary inflammation caused by M. pneumoniae.

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Acknowledgments

This study was supported by the National Research Foundation of Korea (NRF) funded by Ministry of Education, Science and Technology (2012-0000811). H. Kim is grateful to the Brain Korea 21 Project, College of Human Ecology, Yonsei University.

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Authors and Affiliations

  1. Department of Food and Nutrition, Brain Korea 21 Project, College of Human Ecology, Yonsei University, Seoul, 120-749, Korea

    Sang Yong Choi, Joo Weon Lim & Hyeyoung Kim

  2. Department of Infectious Medicine, Kurume University School of Medicine, Kurume, 830-0011, Japan

    Takashi Shimizu & Koichi Kuwano

  3. Department of Microbiology, Hanyang University College of Medicine, Seoul, 133-791, Korea

    Jung Mogg Kim

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  1. Sang Yong Choi
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  2. Joo Weon Lim
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  3. Takashi Shimizu
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Correspondence to Hyeyoung Kim.

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Choi, S.Y., Lim, J.W., Shimizu, T. et al. Reactive oxygen species mediate Jak2/Stat3 activation and IL-8 expression in pulmonary epithelial cells stimulated with lipid-associated membrane proteins from Mycoplasma pneumoniae . Inflamm. Res. 61, 493–501 (2012). https://doi.org/10.1007/s00011-012-0437-7

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  • DOI: https://doi.org/10.1007/s00011-012-0437-7

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