@2024 Afarand., IRAN
ISSN: 2008-2630 Iranian Journal of War & Public Health 2014;6(4):171-176
ISSN: 2008-2630 Iranian Journal of War & Public Health 2014;6(4):171-176
Evaluation of TLR4 Gene Expression in Mustard Gas Injured Persons’ Lungs
ARTICLE INFO
Article Type
Original ResearchAuthors
Ghaffarpour S. (1 )Ghazanfari T. (* )
Salehi I. (2 )
Askary N. (3 )
Mir Afshariyeh A. (4 )
Faghihzadeh E. (1 )
(* ) Immunoregulation Research Center, Shahed University, Tehran, Iran
(1 ) Immunoregulation Research Center, Shahed University, Tehran, Iran
(2 ) Immunology Department, Medicine Faculty, Tehran University, Tehran, Iran
(3 ) Biology Department, Sciences Faculty, Shahid Bahonar University of Kerman, Kerman, Iran
(4 ) Pathology Department, Medicine Faculty, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Correspondence
Article History
Received: July 9, 2014Accepted: August 25, 2014
ePublished: September 20, 2014
ABSTRACT
Aims
Recurrent respiratory infections are one of the clinical protests of those with delayed
complications of exposure to mustard gas. Studies Show that inflammatory mediators have
been changed in serum, sputum and Broncho-Alveolar Lavage (BAL) samples of chemical
victims. Regarding TLR4 importance in inflammation and response to infections, this study
aimed to investigate TLR4 gene expression in chemical victims’ lung tissues.
Materials & Methods In this case-control study, paraffin blocks of lung tissue samples were collected from 28 veterans exposed to mustard gas with delayed pulmonary complications (cases group) and 9 pulmonary samples from the subjects with no mustard exposure (controls group) from the archive of some hospitals during 2013-2014. Slides samples from lung paraffin blocks stained with Hematoxylin and Eosin (H&E) were prepared. Real-time PCR was used to evaluate RNA expression. ΔΔCT median was used to measure the relative genes expression. Comparison of data was done using Mann-Whitney test by SPSS 21 software.
Findings In case group, 12 patients (42.8%) were diagnosed with constructive bronchiolitis, 8 patients (28.6%) with respiratory bronchiolitis and 8 patients (28.6%) with other disease and in control group, 4 patients (44.4%) with constructive bronchiolitis, 1 patient (11.2%) with respiratory bronchiolitis and 4 patients (44.4%) were diagnosed with other diseases. Case group ΔΔCT median was 5.28±3.58cyc and that of control group was 5.81±3.29cyc that gad no significant difference (p>0.05).
Conclusion As there is pathological similarity in lung tissues of both groups, it seems that TLR4 gene expression undergoes same changes.
Materials & Methods In this case-control study, paraffin blocks of lung tissue samples were collected from 28 veterans exposed to mustard gas with delayed pulmonary complications (cases group) and 9 pulmonary samples from the subjects with no mustard exposure (controls group) from the archive of some hospitals during 2013-2014. Slides samples from lung paraffin blocks stained with Hematoxylin and Eosin (H&E) were prepared. Real-time PCR was used to evaluate RNA expression. ΔΔCT median was used to measure the relative genes expression. Comparison of data was done using Mann-Whitney test by SPSS 21 software.
Findings In case group, 12 patients (42.8%) were diagnosed with constructive bronchiolitis, 8 patients (28.6%) with respiratory bronchiolitis and 8 patients (28.6%) with other disease and in control group, 4 patients (44.4%) with constructive bronchiolitis, 1 patient (11.2%) with respiratory bronchiolitis and 4 patients (44.4%) were diagnosed with other diseases. Case group ΔΔCT median was 5.28±3.58cyc and that of control group was 5.81±3.29cyc that gad no significant difference (p>0.05).
Conclusion As there is pathological similarity in lung tissues of both groups, it seems that TLR4 gene expression undergoes same changes.
CITATION LINKS
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[9]Pourfarzam S, Yaraee R, Hassan ZM, Yarmohammadi ME, Faghihzadeh S, Soroush MR, et al. Chemokines, MMP-9 and PMN elastase in spontaneous sputum of sulfur mustard exposed civilians: Sardasht-Iran Cohort Study. Int Immunopharmacol. 2013;17(3):958-63.
[10]Yaraee R, Ghazanfari T, Ebtekar M, Ardestani SK, Rezaei A, Kariminia A, et al. Alterations in serum levels of inflammatory cytokines (TNF, IL-1alpha, IL-1beta and IL-1Ra) 20 years after sulfur mustard exposure: Sardasht-Iran cohort study. Int Immunopharmacol. 2009;9(13-14):1466-70.
[11]Yaraee R, Ghazanfari T, Faghihzadeh S, Mostafaie A, Soroush MR, Inai K, et al. Alterations in the serum levels of soluble L, P and E-selectin 20 years after sulfur mustard exposure: Sardasht-Iran Cohort Study. Int Immunopharmacol. 2009;9(13-14):1477-81.
[12]Kiani A, Mostafaie A, Shirazi FH, Ghazanfari T. Serum profiles of matrix metalloproteinases and their tissue inhibitors in long-term pulmonary complication induced by sulfur mustard: Sardasht-Iran Cohort Study (SICS). Int Immunopharmacol. 2013;17(3):964-7.
[13]Yaraee R, Hassan ZM, Pourfarzam S, Rezaei A, Faghihzadeh S, Ebtekar M, et al. Fibrinogen and inflammatory cytokines in spontaneous sputum of sulfur-mustard-exposed civilians--Sardasht-Iran Cohort Study. Int Immunopharmacol. 2013;17(3):968-73.
[14]Ghanei M, Adibi I. Clinical Review of Mustard Lung. IJMS. 2007;32(2):58-65.
[15]Panahi Y, Ghanei M, Aslani J, Mojtahedzadeh M, Sarhangnejad R, Barkhordari A. Evaluation of microbial resistance to antibiotics in patients with chronic pulmonary lesions due to chemical agents and non-chemical agents. J Mil Med. 2004;6(3):181-6. [Persian]
[16]Emad A, Emad Y. Levels of cytokine in bronchoalveolar lavage (BAL) fluid in patients with pulmonary fibrosis due to sulfur mustard gas inhalation. J Interferon Cytokine Res. 2007;27(1):38-43.
[17]Joeong E, Lee J. Intrinsic and extrinsic regulation of innate immune receptors. Yonsei Med J. 2011;52(3):379-92.
[18]Takeda K, Kaisho T, Akira Sh. Toll-like receptors. Ann Rev Immunol. 2003;21:335-76.
[19]Plummer LE, Smiley-Jewell S, Pinkerton KE. Impact of air pollution on lung inflammation and the role of Toll-like receptors. Int J Interferon Cytokine Mediator Res. 2012;4:43-57.
[20]Gerecke DR, Chen M, Isukapalli SS, Gordon MK, Chang YC, Tong W, et al. Differential gene expression profiling of mouse skin after sulfur mustard exposure: Extended time response and inhibitor effect. Toxicol Appl Pharmacol. 2009;234(2):156-65.
[21]Ghaffarpour S, Ghazanfari T. Correlation between MMP9 and MMP9/ TIMPs complex with pulmonary function in sulfur mustard exposed civilians: Sardasht-Iran Cohort Study. Iran J Immunol. 2014;11:458-9.
[22]Bezemer GF, Sagar S, van Bergenhenegouwen J, Georgiou NA, Garssen J, Kraneveld AD, et al. Dual role of Toll-like receptors in asthma and chronic obstructive pulmonary disease. Pharmacol Rev. 2012;64(2):337-58.
[23]Go H, Koh J, Kim HS, Jeon YK, Chung DH. Expression of toll-like receptor 2 and 4 is increased in the respiratory epithelial cells of chronic idiopathic interstitial pneumonia patients. Respir Med. 2014;108(5):783-92.
[24]Grossman EJ, Shilling RA. Bronchiolitis obliterans in lung transplantation: The good, the bad, and the future. Transl Res. 2009;153(4):153-65.
[25]Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402-8.
[26]Chaudhuri N, Dower SK, Whyte MK, Sabroe I. Toll-like receptors and chronic lung disease. Clin Sci. 2005;109(2):125-33.
[27]Ghanei M, Tazelaar HD, Chilosi M, Harandi AA, Peyman M, Akbari HM, Shamsaei H, et al. An international collaborative pathologic study of surgical lung biopsies from mustard gas-exposed patients. Respir Med. 2008;102(6):825-30
[28]Garantziotis S, Palmer S, Snyder L, Ganous T, Chen B, Wang T, et al. Alloimmune lung injury induced by local innate immune activation through inhaled lipopolysaccharide. Transplantation. 2007;84(8):1012-9.
[29]Tesar BM, Jiang D, Liang J, Palmer SM, Noble PW, Goldstein DR. The Role of Hyaluronan Degradation Products as Innate Alloimmune Agonists. Am J Transplant. 2006;6(11):2622-35.
[30]Pace E, Ferraro M, Minervini MI, Vitulo P, Pipitone L, Chiappara G, et al. Beta defensin-2 is reduced in central but not in distal airways of smoker COPD patients. PLoS One. 2012;7(3):e3360.
[31]Nadigel J, Préfontaine D, Baglole CJ, Maltais F, Bourbeau J, Eidelman DH, et al. Cigarette smoke increases TLR4 and TLR9 expression and induces cytokine production from CD8+ T cells in chronic obstructive pulmonary disease. Respir Res. 2011;12(1):149.
[2]Ghanei M, Chilosi M, Hosseini Akbari HM, Motiei-Langroudi R, Amini Harandi A, Shamsaei H, et al. Use of immunohistochemistry techniques in patients exposed to sulphur mustard gas. Pathol Res Int. 2011. Available From: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3138111.
[3]Ghanei M, Harandi AA. Molecular and cellular mechanism of lung injuries due to exposure to sulfur mustard: A review. Inhal Toxicol. 2011;23(7):363-71.
[4]Razavi SM, Ghanei M, Salamati P, Safiabadi M. Long-term effects of mustard gas on respiratory system of Iranian veterans after Iraq-Iran war: A review. Chinese J Traumatol. 2013;16(3):163-8.
[5]Razavi SM, Ghanei M, Salamati P, Safiabadi M. Long-term effects of mustard gas on respiratory system of Iranian veterans after Iraq-Iran war: A review. Chinese J Traumatol. 2013;16(3):163-8.
[6]Rowell M, Kehe K, Balszuweit F, Thiermann H. The chronic effects of sulfur mustard exposure. Toxicology. 2009;263(1):9-11.
[7]Ghazanfari T, Yaraee R, Kariminia A, Ebtekar M, Faghihzadeh S, Vaez-Mahdavi MR, et al. Alterations in the serum levels of chemokines 20 years after sulfur mustard exposure: Sardasht-Iran Cohort Study. Int Immunopharmacol. 2009;9(13-14):1471-6.
[8]Pourfarzam S, Ghazanfari T, Yaraee R, Ghasemi H, Hassan ZM, Faghihzadeh S, et al. Serum levels of IL-8 and IL-6 in the long term pulmonary complications induced by sulfur mustard: Sardasht-Iran Cohort Study. Int Immunopharmacol. 2009;9(13-14):1482-8.
[9]Pourfarzam S, Yaraee R, Hassan ZM, Yarmohammadi ME, Faghihzadeh S, Soroush MR, et al. Chemokines, MMP-9 and PMN elastase in spontaneous sputum of sulfur mustard exposed civilians: Sardasht-Iran Cohort Study. Int Immunopharmacol. 2013;17(3):958-63.
[10]Yaraee R, Ghazanfari T, Ebtekar M, Ardestani SK, Rezaei A, Kariminia A, et al. Alterations in serum levels of inflammatory cytokines (TNF, IL-1alpha, IL-1beta and IL-1Ra) 20 years after sulfur mustard exposure: Sardasht-Iran cohort study. Int Immunopharmacol. 2009;9(13-14):1466-70.
[11]Yaraee R, Ghazanfari T, Faghihzadeh S, Mostafaie A, Soroush MR, Inai K, et al. Alterations in the serum levels of soluble L, P and E-selectin 20 years after sulfur mustard exposure: Sardasht-Iran Cohort Study. Int Immunopharmacol. 2009;9(13-14):1477-81.
[12]Kiani A, Mostafaie A, Shirazi FH, Ghazanfari T. Serum profiles of matrix metalloproteinases and their tissue inhibitors in long-term pulmonary complication induced by sulfur mustard: Sardasht-Iran Cohort Study (SICS). Int Immunopharmacol. 2013;17(3):964-7.
[13]Yaraee R, Hassan ZM, Pourfarzam S, Rezaei A, Faghihzadeh S, Ebtekar M, et al. Fibrinogen and inflammatory cytokines in spontaneous sputum of sulfur-mustard-exposed civilians--Sardasht-Iran Cohort Study. Int Immunopharmacol. 2013;17(3):968-73.
[14]Ghanei M, Adibi I. Clinical Review of Mustard Lung. IJMS. 2007;32(2):58-65.
[15]Panahi Y, Ghanei M, Aslani J, Mojtahedzadeh M, Sarhangnejad R, Barkhordari A. Evaluation of microbial resistance to antibiotics in patients with chronic pulmonary lesions due to chemical agents and non-chemical agents. J Mil Med. 2004;6(3):181-6. [Persian]
[16]Emad A, Emad Y. Levels of cytokine in bronchoalveolar lavage (BAL) fluid in patients with pulmonary fibrosis due to sulfur mustard gas inhalation. J Interferon Cytokine Res. 2007;27(1):38-43.
[17]Joeong E, Lee J. Intrinsic and extrinsic regulation of innate immune receptors. Yonsei Med J. 2011;52(3):379-92.
[18]Takeda K, Kaisho T, Akira Sh. Toll-like receptors. Ann Rev Immunol. 2003;21:335-76.
[19]Plummer LE, Smiley-Jewell S, Pinkerton KE. Impact of air pollution on lung inflammation and the role of Toll-like receptors. Int J Interferon Cytokine Mediator Res. 2012;4:43-57.
[20]Gerecke DR, Chen M, Isukapalli SS, Gordon MK, Chang YC, Tong W, et al. Differential gene expression profiling of mouse skin after sulfur mustard exposure: Extended time response and inhibitor effect. Toxicol Appl Pharmacol. 2009;234(2):156-65.
[21]Ghaffarpour S, Ghazanfari T. Correlation between MMP9 and MMP9/ TIMPs complex with pulmonary function in sulfur mustard exposed civilians: Sardasht-Iran Cohort Study. Iran J Immunol. 2014;11:458-9.
[22]Bezemer GF, Sagar S, van Bergenhenegouwen J, Georgiou NA, Garssen J, Kraneveld AD, et al. Dual role of Toll-like receptors in asthma and chronic obstructive pulmonary disease. Pharmacol Rev. 2012;64(2):337-58.
[23]Go H, Koh J, Kim HS, Jeon YK, Chung DH. Expression of toll-like receptor 2 and 4 is increased in the respiratory epithelial cells of chronic idiopathic interstitial pneumonia patients. Respir Med. 2014;108(5):783-92.
[24]Grossman EJ, Shilling RA. Bronchiolitis obliterans in lung transplantation: The good, the bad, and the future. Transl Res. 2009;153(4):153-65.
[25]Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402-8.
[26]Chaudhuri N, Dower SK, Whyte MK, Sabroe I. Toll-like receptors and chronic lung disease. Clin Sci. 2005;109(2):125-33.
[27]Ghanei M, Tazelaar HD, Chilosi M, Harandi AA, Peyman M, Akbari HM, Shamsaei H, et al. An international collaborative pathologic study of surgical lung biopsies from mustard gas-exposed patients. Respir Med. 2008;102(6):825-30
[28]Garantziotis S, Palmer S, Snyder L, Ganous T, Chen B, Wang T, et al. Alloimmune lung injury induced by local innate immune activation through inhaled lipopolysaccharide. Transplantation. 2007;84(8):1012-9.
[29]Tesar BM, Jiang D, Liang J, Palmer SM, Noble PW, Goldstein DR. The Role of Hyaluronan Degradation Products as Innate Alloimmune Agonists. Am J Transplant. 2006;6(11):2622-35.
[30]Pace E, Ferraro M, Minervini MI, Vitulo P, Pipitone L, Chiappara G, et al. Beta defensin-2 is reduced in central but not in distal airways of smoker COPD patients. PLoS One. 2012;7(3):e3360.
[31]Nadigel J, Préfontaine D, Baglole CJ, Maltais F, Bourbeau J, Eidelman DH, et al. Cigarette smoke increases TLR4 and TLR9 expression and induces cytokine production from CD8+ T cells in chronic obstructive pulmonary disease. Respir Res. 2011;12(1):149.