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The Role of CTGF in Inflammatory Responses Induced by Silica Particles in Human Bronchial Epithelial Cells

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Abstract

Background

Prolonged exposure to crystalline silica leads to persistent pulmonary inflammation and progressive fibrosis. Connective tissue growth factor (CTGF) has emerged as a potent proinflammatory and profibrotic regulator to participate in a variety of chronic inflammatory diseases. However, the role of CTGF in silica-induced pulmonary inflammation remains poorly understood.

Methods

To explore the effect of CTGF on inflammatory responses caused by silica particles, human bronchial epithelial cells (16HBE) were transfected with CTGF siRNA and exposed to silica particles at concentrations of 0, 12.5, 25, 50, 100 μg/ml for 48 h. Intracellular CTGF mRNA and protein expressions were determined by RT-PCR and Western blotting, respectively. The levels of inflammatory cytokines including IL-8, TNF-α, IL-6, IL-1β, IL-17A and TGF-β1 were measured by ELISA kits.

Results

Silica particles induce significantly elevated intracellular CTGF mRNA expression in 16HBE cells in a dose-dependent manner when compared with blank control group (P < 0.05). The secretions of IL-8, TNF-α, IL-6 and IL-17A were also significantly increased by silica particles (P < 0.05). After exposure to 25 or 50 μg/ml silica particles, the expression of intracellular CTGF mRNA was significantly inhibited in 16HBE cells when transfected with CTGF siRNA (P < 0.05). The secreted levels of IL-8, TNF-α, IL-6 and IL-17A induced by silica particles were also significantly lower from CTGF siRNA-transfected cells than that from normal 16HBE cells (P < 0.05).

Conclusion

Inhibition of CTGF gene attenuates silica-induced inflammatory responses in bronchial epithelial cells, suggesting that CTGF could be a pivotal regulator in the development of silica-induced inflammation.

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References

  1. Chen W, Liu Y, Wang H et al (2012) Long-term exposure to silica dust and risk of total and cause-specific mortality in Chinese workers: a cohort study. PLoS Med 9(4):e1001206

    CAS  PubMed  PubMed Central  Google Scholar 

  2. American Thoracic Society Committee of the Scientific Assembly on Environmental and Occupational Health (1997) Adverse effects of crystalline silica exposure. Am J Respir Crit Care Med 155(2):761–768

    Google Scholar 

  3. Liu Y, Zhou Y, Hnizdo E et al (2017) Total and cause-specific mortality risk associated with low-level exposure to crystalline silica: a 44-year cohort study from China. Am J Epidemiol 186(4):481–490

    PubMed  Google Scholar 

  4. Leung CC, Yu IT, Chen W (2012) Silicosis Lancet 379(9830):2008–2018

    CAS  PubMed  Google Scholar 

  5. Braz NF, Carneiro AP, Avelar NC et al (2016) Influence of Cytokines and soluble receptors in the quality of life and functional capacity of workers exposed to silica. J Occup Environ Med 58(3):272–276

    CAS  PubMed  Google Scholar 

  6. Lee JS, Shin JH, Lee JO et al (2010) Serum levels of interleukin-8 and tumor necrosis factor-alpha in coal workers' pneumoconiosis: one-year follow-up study. Saf Health Work 1(1):69–79

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Zhai R, Ge X, Li H et al (2004) Differences in cellular and inflammatory cytokine profiles in the bronchoalveolar lavage fluid in bagassosis and silicosis. Am J Ind Med 46(4):338–344

    CAS  PubMed  Google Scholar 

  8. Rimal B, Greenberg AK, Rom WN (2005) Basic pathogenetic mechanisms in silicosis: current understanding. Curr Opin Pulm Med 11(2):169–173

    PubMed  Google Scholar 

  9. Carneiro PJ, Clevelario AL, Padilha GA et al (2017) Bosutinib therapy ameliorates lung inflammation and fibrosis in experimental silicosis. Front Physiol 8:159

    PubMed  PubMed Central  Google Scholar 

  10. Kawasaki H (2015) A mechanistic review of silica-induced inhalation toxicity. Inhal Toxicol 27(8):363–377

    CAS  PubMed  Google Scholar 

  11. Sellamuthu R, Umbright C, Roberts JR et al (2017) Molecular mechanisms of pulmonary response progression in crystalline silica exposed rats. Inhal Toxicol 29(2):53–64

    CAS  PubMed  Google Scholar 

  12. Ungvari Z, Valcarcel-Ares MN, Tarantini S et al (2017) Connective tissue growth factor (CTGF) in age-related vascular pathologies. Geroscience 39(5–6):491–498

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Holbourn KP, Acharya KR, Perbal B (2008) The CCN family of proteins: structure-function relationships. Trends Biochem Sci 33(10):461–473

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Takigawa M (2003) CTGF/Hcs24 as a multifunctional growth factor for fibroblasts, chondrocytes and vascular endothelial cells. Drug News Perspect 16(1):11–21

    CAS  PubMed  Google Scholar 

  15. Ramazani Y, Knops N, Elmonem MA et al (2018) Connective tissue growth factor (CTGF) from basics to clinics. Matrix Biol 68–69:44–66

    PubMed  Google Scholar 

  16. Tarr JT, Visser TG, Moon JE et al (2017) The pivotal role of CCN2 in mammalian palatogenesis. J Cell Commun Signal 11(1):25–37

    PubMed  Google Scholar 

  17. Lasky JA, Ortiz LA, Tonthat B et al (1998) Connective tissue growth factor mRNA expression is upregulated in bleomycin-induced lung fibrosis. Am J Physiol 275(2):L365–371

    CAS  PubMed  Google Scholar 

  18. Toda N, Mukoyama M, Yanagita M et al (2018) CTGF in kidney fibrosis and glomerulonephritis. Inflamm Regen 38:14

    PubMed  PubMed Central  Google Scholar 

  19. Braig S, Wallner S, Junglas B et al (2011) CTGF is overexpressed in malignant melanoma and promotes cell invasion and migration. Br J Cancer 105(2):231–238

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Donderski R, Szczepanek J, Domagalski K et al (2013) Analysis of relative expression level of VEGF ( vascular endothelial growth factor ), HIF-1alpha ( hypoxia inducible factor 1alpha ) and CTGF ( connective tissue growth factor ) genes in chronic glomerulonephritis (CGN) patients. Kidney Blood Press Res 38(1):83–91

    CAS  PubMed  Google Scholar 

  21. Ubink I, Verhaar ER, Kranenburg O et al (2016) A potential role for CCN2/CTGF in aggressive colorectal cancer. J Cell Commun Signal 10(3):223–227

    PubMed  PubMed Central  Google Scholar 

  22. Kular L, Pakradouni J, Kitabgi P et al (2011) The CCN family: a new class of inflammation modulators? Biochimie 93(3):377–388

    CAS  PubMed  Google Scholar 

  23. Wang X, McLennan SV, Allen TJ et al (2010) Regulation of pro-inflammatory and pro-fibrotic factors by CCN2/CTGF in H9c2 cardiomyocytes. J Cell Commun Signal 4(1):15–23

    PubMed  PubMed Central  Google Scholar 

  24. Sanchez-Lopez E, Rayego S, Rodrigues-Diez R et al (2009) CTGF promotes inflammatory cell infiltration of the renal interstitium by activating NF-kappaB. J Am Soc Nephrol 20(7):1513–1526

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Yao W, Feng FF, Jiao J, et al. (2006) Expression level and significance of TGF-beta1, PDGF, CTGF in serum of patients with pneumoconiosis. Sichuan Da Xue Xue Bao Yi Xue Ban 37(5):754–756

    CAS  PubMed  Google Scholar 

  26. Guo X, Jagannath C, Espitia MG et al (2012) Uptake of silica and carbon nanotubes by human macrophages/monocytes induces activation of fibroblasts in vitro—potential implication for pathogenesis of inflammation and fibrotic diseases. Int J Immunopathol Pharmacol 25(3):713–719

    CAS  PubMed  Google Scholar 

  27. Perkins TN, Shukla A, Peeters PM, et al. (2012) Differences in gene expression and cytokine production by crystalline vs. amorphous silica in human lung epithelial cells. Part Fibre Toxicol 9(1):6.

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Williams LJ, Zosky GR (2019) The inflammatory effect of iron oxide and silica particles on lung epithelial cells. Lung 197(2):199–207

    CAS  PubMed  Google Scholar 

  29. Ovrevik J, Lag M, Holme JA et al (2009) Cytokine and chemokine expression patterns in lung epithelial cells exposed to components characteristic of particulate air pollution. Toxicology 259(1–2):46–53

    CAS  PubMed  Google Scholar 

  30. Chen F, Shi X (2002) NF-kappaB, a pivotal transcription factor in silica-induced diseases. Mol Cell Biochem 234–235(1–2):169–176

    PubMed  Google Scholar 

  31. Hubbard AK, Timblin CR, Shukla A et al (2002) Activation of NF-kappaB-dependent gene expression by silica in lungs of luciferase reporter mice. Am J Physiol Lung Cell Mol Physiol 282(5):L968–975

    CAS  PubMed  Google Scholar 

  32. van Berlo D, Knaapen AM, van Schooten FJ et al (2010) NF-kappaB dependent and independent mechanisms of quartz-induced proinflammatory activation of lung epithelial cells. Part Fibre Toxicol 7:13

    PubMed  PubMed Central  Google Scholar 

  33. Di Giuseppe M, Gambelli F, Hoyle GW et al (2009) Systemic inhibition of NF-kappaB activation protects from silicosis. PLoS ONE 4(5):e5689

    PubMed  PubMed Central  Google Scholar 

  34. Wahab NA, Brinkman H, Mason RM (2001) Uptake and intracellular transport of the connective tissue growth factor: a potential mode of action. Biochem J 359(Pt 1):89–97

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Gao R, Brigstock DR (2005) Activation of nuclear factor kappa B (NF-kappaB) by connective tissue growth factor (CCN2) is involved in sustaining the survival of primary rat hepatic stellate cells. Cell Commun Signal 3:14

    PubMed  PubMed Central  Google Scholar 

  36. Karatas M, Gunduzoz M, Ozis TN et al (2019) Neutrophil to lymphocyte ratio and platelet to lymphocyte ratio as haematological indices of inflammatory response in ceramic workers' silicosis. Clin Respir J 13(3):159–165

    CAS  PubMed  Google Scholar 

  37. Harada A, Sekido N, Akahoshi T et al (1994) Essential involvement of interleukin-8 (IL-8) in acute inflammation. J Leukoc Biol 56(5):559–564

    CAS  PubMed  Google Scholar 

  38. Lin CH, Wang YH, Chen YW et al (2016) Transcriptional and posttranscriptional regulation of CXCL8/IL-8 gene expression induced by connective tissue growth factor. Immunol Res 64(2):369–384

    CAS  PubMed  Google Scholar 

  39. Das MK, Basak S, Ahmed MS et al (2014) Connective tissue growth factor induces tube formation and IL-8 production in first trimester human placental trophoblast cells. Eur J Obstet Gynecol Reprod Biol 181:183–188

    CAS  PubMed  Google Scholar 

  40. Karger A, Fitzner B, Brock P et al (2008) Molecular insights into connective tissue growth factor action in rat pancreatic stellate cells. Cell Signal 20(10):1865–1872

    CAS  PubMed  Google Scholar 

  41. Cooker LA, Peterson D, Rambow J et al (2007) TNF-alpha, but not IFN-gamma, regulates CCN2 (CTGF), collagen type I, and proliferation in mesangial cells: possible roles in the progression of renal fibrosis. Am J Physiol Renal Physiol 293(1):F157–165

    CAS  PubMed  Google Scholar 

  42. Wang Z, Qiu Y, Lu J et al (2013) Connective tissue growth factor promotes interleukin-1beta-mediated synovial inflammation in knee osteoarthritis. Mol Med Rep 8(3):877–882

    CAS  PubMed  Google Scholar 

  43. Chung BH, Kim KW, Kim BM et al (2015) Increase of Th17 Cell Phenotype in Kidney Transplant Recipients with Chronic Allograft Dysfunction. PLoS ONE 10(12):e0145258

    PubMed  PubMed Central  Google Scholar 

  44. Dudas PL, Sague SL, Elloso MM et al (2011) Proinflammatory/profibrotic effects of interleukin-17A on human proximal tubule epithelium. Nephron Exp Nephrol 117(4):e114–123

    CAS  PubMed  Google Scholar 

  45. Okamoto Y, Hasegawa M, Matsushita T et al (2012) Potential roles of interleukin-17A in the development of skin fibrosis in mice. Arthritis Rheum 64(11):3726–3735

    CAS  PubMed  Google Scholar 

  46. Rodrigues-Diez R, Rodrigues-Diez RR, Rayego-Mateos S et al (2013) The C-terminal module IV of connective tissue growth factor is a novel immune modulator of the Th17 response. Lab Invest 93(7):812–824

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was financially supported by grants from the National Natural Scientific Foundation of China (81402659), the China Scholarship Council Fund (201808420128), the Natural Scientific Foundation of Hubei Province (2014CFB811, 2016CFB520) and the Outstanding Project of Young and Middle-aged Team in Wuhan University of Science and Technology (2018TDZ03).

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Authors and Affiliations

  1. Department of Occupational and Environmental Health, School of Public Health, Medical College, Wuhan University of Science and Technology, Wuhan, 430065, Hubei, China

    Ting Zhou, Qimei Yu, Hui Lin, Guoqing Fu, Lu Lei, Yuqin Shi & Ling Zhang

  2. Hubei Province Key Laboratory of Occupational Hazard Identification and Control, Wuhan University of Science and Technology, Wuhan, 430065, Hubei, China

    Ting Zhou, Qimei Yu, Hui Lin, Zhenyu Wang, Guoqing Fu, Lu Lei, Yuqin Shi & Ling Zhang

  3. Department of Physiology, Wayne State University, Detroit, MI, 48201, USA

    Ting Zhou

  4. Department of Basic Medicine, Medical College, Wuhan University of Science and Technology, Wuhan, 430065, Hubei, China

    Zhenyu Wang

  5. Institute of Pathology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, Hubei, China

    Lingzhi Qin

  6. Department of Epidemiology, School of Public Health, Sun Yat-sen University, Guangzhou, 510080, Guangdong, China

    Yuewei Liu

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  1. Ting Zhou
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Zhou, T., Yu, Q., Lin, H. et al. The Role of CTGF in Inflammatory Responses Induced by Silica Particles in Human Bronchial Epithelial Cells. Lung 197, 783–791 (2019). https://doi.org/10.1007/s00408-019-00272-x

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  • DOI: https://doi.org/10.1007/s00408-019-00272-x

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