Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Inflammation Research
Published in: Inflammation Research 5/2012

01-05-2012 | Original Research Paper

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

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

Published in: Inflammation Research | Issue 5/2012

Login to get access

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.
Literature
1.
2.
Sohn MH, Lee KE, Choi SY, Kwon BC, Chang MW, Kim KE. Effect of Mycoplasma pneumoniae lysate on interleukin-8 gene expression in human respiratory epithelial cells. Chest. 2005;128:322–6.PubMedCrossRef
3.
Hallamaa KM, Tang SL, Ficorilli N, Browning GF. Differential expression of lipoprotein genes in Mycoplasma pneumoniae after contact with human lung epithelial cells, and under oxidative and acidic stress. BMC Microbiol. 2008;8:124.PubMedCrossRef
4.
Barreira ER, Souza DC, Goes PF, Bousso A. Septic shock, necrotizing pneumonitis, and meningoencephalitis caused by Mycoplasma pneumoniae in a child: a case report. Clin Pediatr (Phila). 2009;48:320–2.CrossRef
5.
Waites KB, Talkington DF. Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev. 2004;17:697–728.PubMedCrossRef
6.
Shimizu T, Kida Y, Kuwano K. A dipalmitoylated lipoprotein from Mycoplasma pneumoniae activates NF-kappa B through TLR1, TLR2, and TLR6. J Immunol. 2005;175:4641–6.PubMed
7.
Shimizu T, Kida Y, Kuwano K. Triacylated lipoproteins derived from Mycoplasma pneumoniae activate nuclear factor-kappaB through toll-like receptors 1 and 2. Immunology. 2007;121:473–83.PubMedCrossRef
8.
Shimizu T, Kida Y, Kuwano K. Mycoplasma pneumoniae-derived lipopeptides induce acute inflammatory responses in the lungs of mice. Infect Immun. 2008;76:270–7.PubMedCrossRef
9.
Chambaud I, Wroblewski H, Blanchard A. Interactions between mycoplasma lipoproteins and the host immune system. Trends Microbiol. 1999;7:493–9.PubMedCrossRef
10.
Rawadi G, Zugaza JL, Lemercier B, et al. Involvement of small GTPases in Mycoplasma fermentans membrane lipoproteins-mediated activation of macrophages. J Biol Chem. 1999;274:30794–8.PubMedCrossRef
11.
Yang J, Hooper WC, Phillips DJ, Talkington DF. Cytokines in Mycoplasma pneumoniae infections. Cytokine Growth Factor Rev. 2004;15(2–3):157–68.PubMedCrossRef
12.
Yang J, Hooper WC, Phillips DJ, Talkington DF. Regulation of proinflammatory cytokines in human lung epithelial cells infected with Mycoplasma pneumoniae. Infect Immun. 2002;70:3649–55.PubMedCrossRef
13.
Lieberman D, Livnat S, Schlaeffer F, Porath A, Horowitz S, Levy R. IL-1beta and IL-6 in community-acquired pneumonia: bacteremic pneumococcal pneumonia versus Mycoplasma pneumoniae. Infection. 1997;25:90–4.PubMedCrossRef
14.
Hsieh CC, Tang RB, Tsai CH, Chen W. Serum interleukin-6 and tumor necrosis factor-alpha concentrations in children with Mycoplasma pneumoniae. Microbiol Immunol Infect. 2001;34:109–12.
15.
Narita M, Tanaka H, Yamada S, Abe S, Ariga T, Sakiyama Y. Significant role of interleukin-8 in pathogenesis of pulmonary disease due to Mycoplasma pneumoniae infection. Clin Diagn Lab Immunol. 2001;8:1028–30.PubMed
16.
Kim Y, Seo JH, Kim H. β-Carotene and lutein inhibit hydrogen peroxide-induced activation of NF-κB and IL-8 expression in gastric epithelial AGS cells. J Nutr Sci Vitaminol (Tokyo). 2011;57(3):216–23.CrossRef
17.
Rahman I. Oxidative stress, transcription factors and chromatin remodelling in lung inflammation. Biochem Pharmacol. 2002;64:935–42.PubMedCrossRef
18.
Mukaida N, Mahe Y, Matsushima K. Cooperative interaction of nuclear factor-kappa B- and cis-regulatory enhancer binding protein-like factor binding elements in activating the interleukin-8 gene by pro-inflammatory cytokines. J Biol Chem. 1990;265:21128–33.PubMed
19.
Mukaida N, Okamoto S, Ishikawa Y, Matsushima K. Molecular mechanism of interleukin-8 gene expression. J Leukoc Biol. 1994;56:554–8.PubMed
20.
Tsai YT, Chen YH, Chang DM, Chen PC, Lai JH. Janus kinase/signal transducer and activator of transcription 3 signaling pathway is crucial in chemokine production from hepatocytes infected by dengue virus. Exp Biol Med (Maywood). 2011;236:1156–65.CrossRef
21.
Hu X, Chen J, Wang L, Ivashkiv LB. Crosstalk among Jak-STAT, Toll-like receptor, and ITAM-dependent pathways in macrophage activation. J Leukoc Biol. 2007;82:237–43.PubMedCrossRef
22.
Yu H, Pardoll D, Jove R. STATs in cancer inflammation and immunity: a leading role for STAT3. Nat Rev Cancer. 2009;9:798–809.PubMedCrossRef
23.
Yu JH, Kim KH, Kim H. Suppression of IL-1beta expression by the Jak 2 inhibitor AG490 in cerulein-stimulated pancreatic acinar cells. Biochem Pharmacol. 2006;72:1555–62.PubMedCrossRef
24.
Waris G, Turkson J, Hassanein T, Siddiqui A. Hepatitis C virus (HCV) constitutively activates STAT-3 via oxidative stress: role of STAT-3 in HCV replication. J Virol. 2005;79:1569–80.PubMedCrossRef
25.
Dikdan GS, Saba SC, Dela Torre AN, Roth J, Wang S, Koneru B. Role of oxidative stress in the increased activation of signal transducers and activators of transcription-3 in the fatty livers of obese Zucker rats. Surgery. 2004;136:677–85.PubMedCrossRef
26.
Mazière C, Conte MA, Mazière JC. Activation of JAK2 by the oxidative stress generated with oxidized low-density lipoprotein. Free Radic Biol Med. 2001;31:1334–40.PubMedCrossRef
27.
Schieffer B, Luchtefeld M, Braun S, Hilfiker A, Hilfiker-Kleiner D, Drexler H. Role of NAD(P)H oxidase in angiotensin II-induced JAK/STAT signaling and cytokine induction. Circ Res. 2000;87:1195–201.PubMed
28.
Chen B, Wei J, Wang W, et al. Identification of signaling pathways involved in aberrant production of adipokines in adipocytes undergoing oxidative stress. Arch Med Res. 2009;40:241–8.PubMedCrossRef
29.
Kariya C, Chu HW, Huang J, Leitner H, Martin RJ, Day BJ. Mycoplasma pneumoniae infection and environmental tobacco smoke inhibit lung glutathione adaptive responses and increase oxidative stress. Infect Immun. 2008;76:4455–62.PubMedCrossRef
30.
Sun G, Xu X, Wang Y, Shen X, Chen Z, Yang J. Mycoplasma pneumoniae infection induces reactive oxygen species and DNA damage in A549 human lung carcinoma cells. Infect Immun. 2008;76:4405–13.PubMedCrossRef
31.
Shimizu T, Kida Y, Kuwano K. Mycoplasma pneumoniae-derived lipopeptides induce acute inflammatory responses in the lungs of mice. Infect Immun. 2008;76(1):270–7.PubMedCrossRef
32.
Yu JH, Kim KH, Kim DG, Kim H. Diphenyleneiodonium suppresses apoptosis in cerulein-stimulated pancreatic acinar cells. Int J Biochem Cell Biol. 2007;39:2063–75.PubMedCrossRef
33.
Ju KD, Lim JW, Kim KH, Kim H. Potential role of NADPH oxidase-mediated activation of NADPH oxidase-mediated activation of Jak2/Stat3 and mitogen-activated protein kinases and expression of TGF-β1 in the pathophysiology of acute pancreatitis. Inflamm Res. 2011;60:791–800.PubMedCrossRef
34.
Kim YH, Seo JH, Kim H. Beta-carotene and lutein inhibit hydrogen peroxide-induced activation of NF-κB and IL-8 expression in gastric epithelial AGS cells. J Nutr Sci Vitaminol. 2011;57:216–23.PubMedCrossRef
35.
Yu JH, Kim KH, Kim H. SOCS3 and PPAR-gamma ligands inhibit the expression of IL-6 and TGF-beta 1 by regulating JAK1/STAT3 signaling in pancreas. Int J Biochem Cell Biol. 2008;40:677–88.PubMedCrossRef
36.
37.
Chmura K, Lutz RD, Chiba H, et al. Mycoplasma pneumoniae antigens stimulate interleukin-8. Chest. 2003;123:425S.PubMedCrossRef
38.
Trevino JG, Gray MJ, Nawrocki ST, et al. Src activation of Stat3 is an independent requirement from NF-kappaB activation for constitutive IL-8 expression in human pancreatic adenocarcinoma cells. Angiogenesis. 2006;9:101–10.PubMedCrossRef
39.
Vij N, Sharma A, Thakkar M, Sinha S, Mohan RR. PDGF-driven proliferation, migration, and IL-8 chemokine secretion in human corneal fibroblasts involve JAK2-STAT3 signaling pathway. Mol Vis. 2008;14:1020–7.PubMed
40.
Gharavi NM, Alva JA, Mouillesseaux KP, et al. Role of the Jak/Stat pathway in the regulation of interleukin-8 transcription by oxidized phospholipids in vitro and in atherosclerosis in vivo. J Biol Chem. 2007;282:31460–8.PubMedCrossRef
41.
Almagor M, Kahane I, Yatziv S. Role of superoxide anion in host cell injury induced by Mycoplasma pneumoniae infection. A study in normal and trisomy 21 cells. J Clin Invest. 1984;73:842–7.PubMedCrossRef
42.
Almagor M, Kahane I, Gilon C, Yatziv S. Protective effects of the glutathione redox cycle and vitamin E on cultured fibroblasts infected by Mycoplasma pneumoniae. Infect Immun. 1986;52:240–4.PubMed
43.
Strassheim D, Riddle SR, Burke DL, Geraci MW, Stenmark KR. Prostacyclin inhibits IFN-gamma-stimulated cytokine expression by reduced recruitment of CBP/p300 to STAT1 in a SOCS-1-independent manner. J Immunol. 2009;183(11):6981–8.PubMedCrossRef
44.
Wertheimer AM, Polyak SJ, Leistikow R, Rosen HR. Engulfment of apoptotic cells expressing HCV proteins leads to differential chemokine expression and STAT signaling in human dendritic cells. Hepatology. 2007;45(6):1422–32.PubMedCrossRef
45.
West AP, Brodsky IE, Rahner C, et al. TLR signaling augments macrophage bactericidal activity through mitochondrial ROS. Nature. 2011;472:476–80.PubMedCrossRef
46.
Scharf S, Hippenstiel S, Flieger A, Suttorp N, N’Guessan PD. Induction of human β-defensin-2 in pulmonary epithelial cells by Legionella pneumophila: involvement of TLR2 and TLR5, p38 MAPK, JNK, NF-κB, and AP-1. Am J Physiol Lung Cell Mol Physiol. 2010;298(5):L687–95.PubMedCrossRef
47.
Wu TT, Chen TL, Loon WS, Tai YT, Cherng YG, Chen RM. Lipopolysaccharide stimulates syntheses of toll-like receptor 2 and surfactant protein-A in human alveolar epithelial A549 cells through upregulating phosphorylation of MEK1 and ERK1/2 and sequential activation of NF-κB. Cytokine. 2011;55(1):40–7.PubMedCrossRef
48.
Ivanov VN, Ghandhi SA, Zhou H, et al. Radiation response and regulation of apoptosis induced by a combination of TRAIL and CHX in cells lacking mitochondrial DNA: a role for NF-κB–STAT3-directed gene expression. Exp Cell Res. 2011;317(11):1548–66.PubMedCrossRef
49.
Funakoshi-Tago M, Tago K, Sato Y, Tominaga S, Kasahara T. JAK2 is an important signal transducer in IL-33-induced NF-κB activation. Cell Signal. 2011;23(2):363–70.PubMedCrossRef
50.
Li X, Kim KW, Cho ML, et al. IL-23 induces receptor activator of NF-kappaB ligand expression in fibroblast-like synoviocytes via STAT3 and NF-kappaB signal pathways. Immunol Lett. 2010;127(2):100–7.PubMedCrossRef
51.
Helin E, Vainionpää R, Hyypiä T, Julkunen I, Matikainen S. Measles virus activates NF-kappa B and STAT transcription factors and production of IFN-alpha/beta and IL-6 in the human lung epithelial cell line A549. Virology. 2001;290(1):1–10.PubMedCrossRef
52.
Morris PE, Bernard GR. Significance of glutathione in lung diseases and implications for therapy. Am J Med Sci. 1994;307:119–27.PubMedCrossRef
53.
Gressier B, Cabanis A, Lebegue S. Decrease of hypochlorous acid and hydroxyl radical generated by stimulated human neutrophils: comparison in vitro of some thiol-containing drugs. Methods Find Exp Clin Pharmacol. 1994;16:9–13.PubMed
54.
Quan Y, Jiang CT, Xue B, Zhu SG, Wang X. High glucose stimulates TNFα and MCP-1 expression in rat microglia via ROS and NF-κB pathways. Acta Pharmacol Sin. 2011;32(2):188–93.PubMedCrossRef
55.
Mata M, Morcillo E, Gimeno C, Cortijo J. N-acetyl-l-cysteine (NAC) inhibit mucin synthesis and pro-inflammatory mediators in alveolar type II epithelial cells infected with influenza virus A and B and with respiratory syncytial virus (RSV). Biochem Pharmacol. 2011;82(5):548–55.PubMedCrossRef
56.
Petrescu BC, Gurzu B, Iancu RI, et al. Apelin effects on lipopolysaccharide-increased pulmonary permeability in rats. Rev Med Chir Soc Med Nat Iasi. 2010;114(1):163–9.PubMed
57.
Chu SH, Kim H, Seo JY, Lim JW, Mukaida N, Kim KH. Role of NF-kB and AP-1 on Helicobacter pylori-induced IL-8 expression in AGS cells. Dig Dis Sci. 2003;48(2):257–65.PubMedCrossRef
58.
Roebuck KA. Oxidant stress regulation of IL-8 and ICAM-1 gene expression: different activation and binding of the transcription factors AP-1 and NF-κB. Int J Mol Med. 1999;4:223–30.PubMed
Metadata
Title
Reactive oxygen species mediate Jak2/Stat3 activation and IL-8 expression in pulmonary epithelial cells stimulated with lipid-associated membrane proteins from Mycoplasma pneumoniae
Authors
Sang Yong Choi
Joo Weon Lim
Takashi Shimizu
Koichi Kuwano
Jung Mogg Kim
Hyeyoung Kim
Publication date
01-05-2012
Publisher
SP Birkhäuser Verlag Basel
Published in
Inflammation Research / Issue 5/2012
Print ISSN: 1023-3830
Electronic ISSN: 1420-908X
DOI
https://doi.org/10.1007/s00011-012-0437-7

Other articles of this Issue 5/2012

Inflammation Research 5/2012 Go to the issue

Original Research Paper

Histamine modulates salivary secretion and diminishes the progression of periodontal disease in rat experimental periodontitis

Original Research Paper

Effects of 1,25-(OH)2D3 on the expressions of vitamin D receptor, STAT5 and cytoskeletal rearrangement in human monocytes incubated with sera from type 2 diabetes patients and diabetic nephropathy patients with uremia

Original Research Paper

HSP90 inhibition by 17-DMAG reduces inflammation in J774 macrophages through suppression of Akt and nuclear factor-κB pathways

Original Research Paper

Suppressive effect of an orally active MEK1/2 inhibitor in two different animal models for rheumatoid arthritis: a comparison with leflunomide

Original Research Paper

Prevention of 1-palmitoyl lysophosphatidylcholine-induced inflammation by polyunsaturated acyl lysophosphatidylcholine

Short Communication

The association between the functional PTPN22 1858 C/T and MIF −173 C/G polymorphisms and juvenile idiopathic arthritis: a meta-analysis
Obesity Clinical Trial Summary

At a glance: The STEP trials

Read more

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

Watch now

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits