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

Sang Yong Choi, Joo Weon Lim, Takashi Shimizu, Koichi Kuwano, Jung Mogg Kim, Hyeyoung Kim

Research output: Contribution to journalArticle

21 Citations (Scopus)

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 transcriptionpolymerase 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. copyright

Original languageEnglish
Pages (from-to)493-501
Number of pages9
JournalInflammation Research
Volume61
Issue number5
DOIs
Publication statusPublished - 2012 May 1

Fingerprint

Mycoplasma Pneumonia
Transducers
Interleukin-8
Reactive Oxygen Species
Membrane Proteins
Epithelial Cells
Lipids
Lung
Acetylcysteine
Mycoplasma pneumoniae
Antioxidants
DNA
Electrophoretic Mobility Shift Assay
Pneumonia
Western Blotting
Enzyme-Linked Immunosorbent Assay
Phosphorylation
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Immunology
  • Pharmacology

Cite this

@article{d9f9bf7cee9e4c4993eaf9fd38742d9a,
title = "Reactive oxygen species mediate Jak2/Stat3 activation and IL-8 expression in pulmonary epithelial cells stimulated with lipid-associated membrane proteins from Mycoplasma pneumonia",
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 transcriptionpolymerase 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. copyright",
author = "Choi, {Sang Yong} and Lim, {Joo Weon} and Takashi Shimizu and Koichi Kuwano and Kim, {Jung Mogg} and Hyeyoung Kim",
year = "2012",
month = "5",
day = "1",
doi = "10.1007/s00011-012-0437-7",
language = "English",
volume = "61",
pages = "493--501",
journal = "Inflammation Research",
issn = "1023-3830",
publisher = "Birkhauser Verlag Basel",
number = "5",

}

Reactive oxygen species mediate Jak2/Stat3 activation and IL-8 expression in pulmonary epithelial cells stimulated with lipid-associated membrane proteins from Mycoplasma pneumonia. / Choi, Sang Yong; Lim, Joo Weon; Shimizu, Takashi; Kuwano, Koichi; Kim, Jung Mogg; Kim, Hyeyoung.

In: Inflammation Research, Vol. 61, No. 5, 01.05.2012, p. 493-501.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Reactive oxygen species mediate Jak2/Stat3 activation and IL-8 expression in pulmonary epithelial cells stimulated with lipid-associated membrane proteins from Mycoplasma pneumonia

AU - Choi, Sang Yong

AU - Lim, Joo Weon

AU - Shimizu, Takashi

AU - Kuwano, Koichi

AU - Kim, Jung Mogg

AU - Kim, Hyeyoung

PY - 2012/5/1

Y1 - 2012/5/1

N2 - 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 transcriptionpolymerase 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. copyright

AB - 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 transcriptionpolymerase 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. copyright

UR - http://www.scopus.com/inward/record.url?scp=84863108461&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84863108461&partnerID=8YFLogxK

U2 - 10.1007/s00011-012-0437-7

DO - 10.1007/s00011-012-0437-7

M3 - Article

C2 - 22270622

AN - SCOPUS:84863108461

VL - 61

SP - 493

EP - 501

JO - Inflammation Research

JF - Inflammation Research

SN - 1023-3830

IS - 5

ER -