Evaluation of the LTBP1 and Smad6 Genes Expression in Lung Tissue of Sulfur Mustard-exposed Individuals with Long-term Pulmonary Complications
Abstract
Sulfur mustard (SM) exposure injures different organs such as the lungs and leads to short and long term complications Transforming growth factor beta (TGF-β) has the main role in altering fibroblast activities linked to airways remodeling. Latency TGF beta binding proteins 1 (LTBP1 facilitates localization of TGF-β in the extracellular matrix. Mothers against decapentaplegic homolog 6 (Smad6) negatively regulates TGF-β signaling, thus establishing a main negative feedback loop. In this study, we investigated the expression of LTBP1 and Smad6 in the lung tissues of SM-exposed and control individuals. Lung formalin-fixed paraffin-embedded (FFPE) blocks of SM-exposed (20 samples) and control groups (20 samples) were collected from archival pathology department of several general hospitals. The total mRNA of lung FFPE tissues was extracted. Quality of the extracted mRNA was evaluated by an Agilent Bio analyzer and RNA was quantified using a Nano Drop. LTBP1 and Smad6 expression levels were evaluated by real-time PCR. LTBP1 expression levels did not change between the two groups (p=0.626), howeverSmad6 expression levels were significantly higher (2.6 fold) in SM-exposed individuals compared to the control group (p=0.001). Our results revealed that Smad6 may be involved in lung tissue remodeling process in SM-exposed patients. Smad6 regulates fibrotic alterations in lung tissue and its function as negative feedback mechanisms in TGF-β.
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26. www.biotools.nubic.northwestern.edu/OligoCalc
27. Eghtedardoost M, Hassan ZM, Askari N, Sadeghipour A, Naghizadeh MM, Ghafarpour S, et al. The delayed effect of mustard gas on housekeeping gene expression in lung biopsy of chemical injuries. BiochemBiophys Rep 2017; 11:27-32.
28. Pfaffi MW. A new mathematical model for relative quantification in real time RT-PCR. Nucleic Acids Res 2001; 29(9):e45.
29. Hynes, Richard O. The extracellular matrix: not just pretty fibrils. Science 2009; 326(5957):1216-9.
30. Ezzie ME, Crawford M, Cho J-H, Orellana R, Zhang S, Gelinas R, et al. Gene expression networks in COPD: microRNA and mRNA regulation. Thorax 2012; 67(2):122-31.
31. Aghanouri R, Ghanei M, Aslani J, Keivani-Amine H, Rastegar F, Karkhane A. Fibrogenic cytokine levels in bronchoalveolar lavage aspirates 15 years after exposure to sulfur mustard. Am J Physiol Lung Cell MolPhysiol 2004; 287(6):1160-4.
32. Weinberger B, Laskin JD, Sunil VR, Sinko PJ, Heck DE, Laskin DL. Sulfur mustard-induced pulmonary injury: Therapeutic approaches to mitigating toxicity. PulmPharmacolTher 2011; 24(1):92-9.
33. Ghazanfari T, Yaraee R, Kariminia A, Ebtekar M, Faghihzadeh S, Vaez-Mahdavi MR, et al. Alterations in the serum levels of chemokines 20years after sulfur mustard exposure: Sardasht-Iran Cohort Study.IntImmunopharmacol 2009; 9(13-14):1471-6.
34. Zhang X, Zheng H, Zhang H, Ma W, Wang F, Liu C, et al. Increased interleukin (IL)-8 and decreased IL-17production in chronic obstructive pulmonary disease (COPD) provoked by cigarette smoke. Cytokine 2011; 56(3):717-25.
35. Krafft E, Lybaert P. Transforming Growth Factor Beta 1 Activation, Storage, and Signaling Pathways in Idiopathic Pulmonary Fibrosis in Dogs. J Vet Intern Med 2014; 28(6):1666–75.
2. Hassan ZM, Ebtekar M, Ghanei M, Taghikhani M, Daloii MRN, Ghazanfari T. Immunobiological consequences of sulfur mustard contamination. Iran J Allergy Asthma Immunol 2006; 5(3):101-8.
3. Ghasemi H, Ghazanfari T, Babaei M, Soroush MR, Yaraee R, Ghassemi-Broumand M, et al. Long-term ocular complications of sulfur mustard in the civilian victims of Sardasht, Iran. Cutan Ocul Toxicol 2008; 27(4):317-26.
4. Balali-Mood M, Mousavi S, Balali-Mood B. Chronic health effects of sulphur mustard exposure with special reference to Iranian veterans. Emerg Health Threats J 2008; 1(1):7e.
5. Najafi A, Ghanei M, Jamalkandi SA. Airway remodeling: Systems biology approach, from bench to bedside. Technol Health Care 2016; 24(6);811-9.
6. Pourfarzam S, Ghazanfari T, Merasizadeh J, Ghanei M, Azimi G, Araghizadeh H, et al. Long-term pulmonary complications in sulfur mustard victims of Sardasht, Iran. Toxin Reviews. 2009;28(1):8-13.
7. Khazdair MR, Boskabady MH, Ghorani V. Respiratory effects of sulfur mustard exposure, similarities and differences with asthma and COPD. InhalToxicol 2015; 27(14):731-44.
8. Annes JP, Chen Y, Munger JS, Rifkin DB. Integrin alphaVbeta6-mediated activation of latent TGF-beta requires the latent TGF-beta binding protein-1. J Cell Biol 2004; 165(5):723–34.
9. Ihn H, Yamane K, Kubo M, Tamaki K. Blockade of endogenous TGF Beta singnaling prevents up-regulate collagen synthesis in scleroderma fibroblasts; association with increased expression of transforming growth factor receptors. Arthritis Rheum 2001; 44:474-80.
10. Pha Phan SH. Fibroblast phenotypes in pulmonaryfibrosis. Am J Respir Cell MolBiol 2003; 29 (3 Suppl):87–92.
11. Gordon KJ, Blobe GC. Role of transforming growth factor beta in human disease. N Engl J Med 2000; 342(18):1350-8.
12. Tylor A. review of the activation of TGF-Beta in immunity. J LeukocBiol 2009; 85(1):29-33.
13. Massam-Wu T, Chiu M, Choudhury R, Chaudhry SS, Baldwin AK, McGovern A, et al. Assembly of fibrillinmicrofibrils governs extracellular deposition of latent TGF beta. J Cell Sci 2010; 123(14):3006-11.
14. Tsuji T, Okada F, Yamaguchi K, Nakamura T. Molecular cloning of the large subunit of transforming growth factor type beta masking protein and expression of the mRNA in various rat tissues. Proc Natl AcadSci U S A 1990; 87(22):8835–9.
15. Miyazono K, Olofsson A, Colosetti P, Heldin CH. A role of the latent TGF-beta 1-binding protein in the assembly and secretion of TGF-beta 1. EMBO J 1991; 10(5):1091–101.
16. Shi M, Zhu J, Wang R, Chen X, Mi L, Walz T, et al. LatentTGF-beta structure and activation. Nature 2011; 474:343–9.
17. Ebrahimi M, Roudkenar M H, ImanFooladi AA, Halabian R, Ghanei M, Kondo H , et al. Discrepancy between MmRNA and Protein Expression of Neutrophil Gelatinase-Associated Lipocalin in Bronchial Epithelium Induced by SulfurMustard. J Biomed and biotech. 2010..
18. Derynck R, Zhang YE. Smad-dependent and Smad-independent pathways in TGF-β family signalling. Nature 2003; 425(6958):577.
19. . Miyazawa K, Shinozaki M, Hara T, Furuya T, Miyazono K. Two major Smad pathways in TGFb superfamily signaling. Genes to Cells 2002; 7(12):1191-204.
20. Massagué J. How cells read TGF-b signals. Nature Rev. Mol Cell Biol 2000; 1(3):169–78.
21. Moustakas, A, Souchelnytskyi S. Smad regulation in TGF-b signal transduction. J Cell Sci 2001; 114:4359–69.
22. Zarin A, Behmanesh M, Tavallaei M, Shohrati M, Ghanei M. Overexpression of transforming growth factor (TGF)-β1 and TGF-β3 genes in lung of toxic-inhaled patients. Exp Lung Res 2010; 36(5):284-91.
23. 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.
24. . Leppäranta O. Sens C. Regulation of TGF-β storage and activation in the human idiopathic pulmonary fibrosis lung. Cell Tissue Res 2012; 348(3):491–503.
25. Springer J, Groneberg A, Fisher A. SMAD-signaling in chronic obstructive pulmonary disease. transcriptional down-regulation of inhibitory SMAD 6 and 7 by cigarette smoke. BiolChem 2004; 385(7):649-53.
26. www.biotools.nubic.northwestern.edu/OligoCalc
27. Eghtedardoost M, Hassan ZM, Askari N, Sadeghipour A, Naghizadeh MM, Ghafarpour S, et al. The delayed effect of mustard gas on housekeeping gene expression in lung biopsy of chemical injuries. BiochemBiophys Rep 2017; 11:27-32.
28. Pfaffi MW. A new mathematical model for relative quantification in real time RT-PCR. Nucleic Acids Res 2001; 29(9):e45.
29. Hynes, Richard O. The extracellular matrix: not just pretty fibrils. Science 2009; 326(5957):1216-9.
30. Ezzie ME, Crawford M, Cho J-H, Orellana R, Zhang S, Gelinas R, et al. Gene expression networks in COPD: microRNA and mRNA regulation. Thorax 2012; 67(2):122-31.
31. Aghanouri R, Ghanei M, Aslani J, Keivani-Amine H, Rastegar F, Karkhane A. Fibrogenic cytokine levels in bronchoalveolar lavage aspirates 15 years after exposure to sulfur mustard. Am J Physiol Lung Cell MolPhysiol 2004; 287(6):1160-4.
32. Weinberger B, Laskin JD, Sunil VR, Sinko PJ, Heck DE, Laskin DL. Sulfur mustard-induced pulmonary injury: Therapeutic approaches to mitigating toxicity. PulmPharmacolTher 2011; 24(1):92-9.
33. Ghazanfari T, Yaraee R, Kariminia A, Ebtekar M, Faghihzadeh S, Vaez-Mahdavi MR, et al. Alterations in the serum levels of chemokines 20years after sulfur mustard exposure: Sardasht-Iran Cohort Study.IntImmunopharmacol 2009; 9(13-14):1471-6.
34. Zhang X, Zheng H, Zhang H, Ma W, Wang F, Liu C, et al. Increased interleukin (IL)-8 and decreased IL-17production in chronic obstructive pulmonary disease (COPD) provoked by cigarette smoke. Cytokine 2011; 56(3):717-25.
35. Krafft E, Lybaert P. Transforming Growth Factor Beta 1 Activation, Storage, and Signaling Pathways in Idiopathic Pulmonary Fibrosis in Dogs. J Vet Intern Med 2014; 28(6):1666–75.
| Files | ||
| Issue | Vol 18, No 5 (2019) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijaai.v18i5.1895 | |
| PMID | 32245290 | |
| Keywords | ||
| Latency TGF beta binding proteins 1 (LTBP1) Mothers against decapentaplegic homolog 6 (SMAD6) Sulfur mustard Transforming growth factor β | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Mami S, Ghaffarpour S, Faghihzadeh S, Ghazanfari T. Evaluation of the LTBP1 and Smad6 Genes Expression in Lung Tissue of Sulfur Mustard-exposed Individuals with Long-term Pulmonary Complications. Iran J Allergy Asthma Immunol. 2019;18(5):473-478.
