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Tannic acid prevents macrophage-induced pro-fibrotic response in lung epithelial cells via suppressing TLR4-mediated macrophage polarization

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Abstract

Background

Polarized macrophages induce fibrosis through multiple mechanisms, including a process termed epithelial-to-mesenchymal transition (EMT). Mesenchymal cells contribute to the excessive accumulation of fibrous connective tissues, leading to organ failure. This study was aimed to investigate the effect of tannic acid (TA), a natural dietary polyphenol on M1 macrophage-induced EMT and its underlying mechanisms.

Materials

First, we induced M1 polarization in macrophage cell lines (RAW 264.7 and THP-1). Then, the conditioned-medium (CM) from these polarized macrophages was used to induce EMT in the human adenocarcinomic alveolar epithelial (A549) cells. We also analysed the role of TA on macrophage polarization.

Results

We found that TA pre-treated CM did not induce EMT in epithelial cells. Further, TA pre-treated CM showed diminished activation of MAPK in epithelial cells. Subsequently, TA was shown to inhibit LPS-induced M1 polarization in macrophages by directly targeting toll-like receptor 4 (TLR4), thereby repressing LPS binding to TLR4/MD2 complex and subsequent signal transduction.

Conclusion

It was concluded that TA prevented M1 macrophage-induced EMT by suppressing the macrophage polarization possibly through inhibiting the formation of LPS-TLR4/MD2 complex and blockage of subsequent downstream signal activation. Further, our findings may provide beneficial information to develop new therapeutic strategies against chronic inflammatory diseases.

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References

  1. 1.

    Gilbane AJ, Denton CP, Holmes AM. Scleroderma pathogenesis: a pivotal role for fibroblasts as effector cells. Arthritis Res Ther. 2013;15:215.

  2. 2.

    Ramming A, Dees C, Distler JH. From pathogenesis to therapy-Perspective on treatment strategies in fibrotic diseases. Pharmacol Res. 2015;100:93–100.

  3. 3.

    Willis BC, duBois RM, Borok Z. Epithelial origin of myofibroblasts during fibrosis in the lung. Proc Am Thorac Soc. 2006;3:377–82.

  4. 4.

    Iwano M, Plieth D, Danoff TM, Xue C, Okada H, Neilson EG. Evidence that fibroblasts derive from epithelium during tissue fibrosis. J Clin Invest. 2002;110:341–50.

  5. 5.

    Lamouille S, Xu J, Derynck R. Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 2014;15:178–96.

  6. 6.

    Stone RC, Pastar I, Ojeh N, Chen V, Liu S, Garzon KI, et al. Epithelial-mesenchymal transition in tissue repair and fibrosis. Cell Tissue Res. 2016;365:495–506.

  7. 7.

    Braga TT, Agudelo JS, Camara NO. Macrophages during the fibrotic process: M2 as friend and foe. Front Immunol. 2015;25:602.

  8. 8.

    Cao Q, Harris DC, Wang Y. Macrophages in kidney injury, inflammation, and fibrosis. Physiology (Bethesda). 2015;30:183–94.

  9. 9.

    Mantovani A, Biswas SK, Galdiero MR, Sica A, Locati M. Macrophage plasticity and polarization in tissue repair and remodelling. J Pathol. 2013;229:176–85.

  10. 10.

    Shi J, Li Q, Sheng M, Zheng M, Yu M, Zhang L. The role of TLR4 in M1 macrophage-induced epithelial-mesenchymal transition of peritoneal mesothelial cells. Cell Physiol Biochem. 2016;40:1538–48.

  11. 11.

    Scalbert A, Manach C, Morand C, Rémésy C, Jiménez L. Dietary polyphenols and the prevention of diseases. Crit Rev Food Sci Nutr. 2005;45:287–306.

  12. 12.

    Salminen JP, Karonen M. Chemical ecology of tannins and other phenolics: we need a change in approach. Funct Ecol. 2011;25:325–38.

  13. 13.

    Pattarayan D, Sivanantham A, Krishnaswami V, Loganathan L, Palanichamy R, Natesan S, et al. Tannic acid attenuates TGF-β1-induced epithelial-to-mesenchymal transition by effectively intervening TGF-β signaling in lung epithelial cells. J Cell Physiol. 2018;233:2513–25.

  14. 14.

    Pattarayan D, Sivanantham A, Bethunaickan R, Palanichamy R, Rajasekaran S. Tannic acid modulates fibroblast proliferation and differentiation in response to pro-fibrotic stimuli. J Cell Biochem. 2018;119:6732–42.

  15. 15.

    Sivanantham A, Pattarayan D, Bethunaickan R, Kar A, Mahapatra SK, Thimmulappa RK, et al. Tannic acid protects against experimental acute lung injury through down-regulation of TLR4 and MAPK. J Cell Physiol. 2019;234:6463–76.

  16. 16.

    Chai WM, Wei QM, Deng WL, Zheng YL, Chen XY, Huang Q, et al. Anti-melanogenesis properties of condensed tannins from Vigna angularis seeds with potent antioxidant and DNA damage protection activities. Food Funct. 2019;10:99–111.

  17. 17.

    Koval A, Pieme CA, Queiroz EF, Ragusa S, Ahmed K, Blagodatski A, et al. Tannins from Syzygium guineense suppress Wnt signaling and proliferation of Wnt-dependent tumors through a direct effect on secreted Wnts. Cancer Lett. 2018;435:110–20.

  18. 18.

    Gourlay G, Constabel CP. Condensed tannins are inducible antioxidants and protect hybrid polar against oxidative stress. Tree Physiol. 2019;39:345–55.

  19. 19.

    Helmy IM, Azim AM. Efficacy of ImageJ in the assessment of apoptosis. Diag Pathol. 2012;7:15.

  20. 20.

    Wang Y, Su L, Morin MD, Jones BT, Whitby LR, Surakattula MM, et al. TLR4/MD-2 activation by a synthetic agonist with no similarity to LPS. Proc Natl Acad Sci USA. 2016;113:E884–93.

  21. 21.

    Maestro Suite, 2017, Version11.4.011, Schrodinger, LLC, NY.

  22. 22.

    Loganathan L, Muthusamy K. Investigation of drug interaction potentials and binding modes on direct renin inhibitors. A computational modeling studies. Lett Drug Des Discov. 2019;16:919–38.

  23. 23.

    Kawasaki K, Nogawa H, Nishijima M. Identification of mouse MD-2 residues important for forming the cell surface TLR4-MD-2 complex recognized by anti-TLR4-MD-2 antibodies, and for conferring LPS and taxol responsiveness on mouse TLR4 by alanine-scanning mutagenesis. J Immunol. 2003;170:413–20.

  24. 24.

    Park BS, Song DH, Kim HM, Choi BS, Lee H, Lee JO. The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex. Nature. 2009;458:1191–5.

  25. 25.

    Santos AFM, Macedo LJA, Chaves MH, Castaneda ME, Merkoci A, Lima FCA, et al. Hybrid self-assembled materials constituted by ferromagnetic nanoparticles and tannic acid: a theoretical and experimental investigation. J Braz Chem Soc. 2016;27:727–34.

  26. 26.

    Kong J, Yu S. Fourier transform infrared spectroscopic analysis of protein secondary structures. Acta Biochim Biophys Sin. 2007;39:549–59.

  27. 27.

    Deng YR, Liu WB, Lian ZX, Li X, Hou X. Sorafenib inhibits macrophage-mediated epithelial-mesenchymal transition in hepatocellular carcinoma. Oncotarget. 2016;7:38292–305.

  28. 28.

    Wu KQ, Muratore CS, So EY, Sun C, Dubielecka PM, Reginato AM, et al. M1 macrophage-induced endothelial-to-mesenchymal transition promotes infantile hemangioma regression. Am J Pathol. 2017;187:2102–11.

  29. 29.

    Yeung OW, Lo CM, Ling CC, Qi X, Geng W, Li CX, et al. Alternatively activated (M2) macrophages promote tumour growth and invasiveness in hepatocellular carcinoma. J Hepatol. 2015;62:607–16.

  30. 30.

    Li Q, Lv LL, Wu M, Zhang XL, Liu H, Liu BC. Dexamethasone prevents monocyte-induced tubular epithelial-mesenchymal transition in HK-2 cells. J Cell Biochem. 2003;114:632–8.

  31. 31.

    Borthwick LA, Corris PA, Mahida R, Walker A, Gardner A, Suwara M, et al. TNFα from classically activated macrophages accentuates epithelial to mesenchymal transition in obliterative bronchiolitis. Am J Transplant. 2013;13:621–33.

  32. 32.

    Yamada M, Kuwano K, Maeyama T, Hamada N, Yoshimi M, Nakanishi Y, et al. Dual-immunohistochemistry provides little evidence for epithelial-mesenchymal transition in pulmonary fibrosis. Histochem Cell Biol. 2008;129:453–62.

  33. 33.

    Marmai C, Sutherland RE, Kim KK, Dolganov GM, Fang X, Kim SS, et al. Alveolar epithelial cells express mesenchymal proteins in patients with idiopathic pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol. 2011;301:L71–8.

  34. 34.

    Liang H, Gu Y, Li T, Zhang Y, Huangfu L, Hu M, et al. Integrated analyses identify the involvement of microRNA-26a in epithelial-mesenchymal transition during idiopathic pulmonary fibrosis. Cell Death Dis. 2014;5:e1238.

  35. 35.

    Tanjore H, Xu XC, Polosukhin VV, Degryse AL, Li B, Han W, et al. Contribution of epithelial-derived fibroblasts to bleomycin-induced lung fibrosis. Am J Respir Crit Care Med. 2009;180:657–65.

  36. 36.

    Inai T, Kobayashi J, Shibata Y. Claudin-1 contributes to the epithelial barrier function in MDCK cells. Eur J Cell Biol. 1999;78:849–55.

  37. 37.

    Shiozaki A, Bai XH, Shen-Tu G, Moodley S, Takeshita H, Fung SY, et al. Claudin 1 mediates TNFα-induced gene expression and cell migration in human lung carcinoma cells. PLoS One. 2012;7:e38049.

  38. 38.

    Fortier AM, Asselin E, Cadrin M. Keratin 8 and 18 loss in epithelial cancer cells increases collective cell migration and cisplatin sensitivity through claudin1 up-regulation. J Biol Chem. 2013;288:11555–71.

  39. 39.

    Grund EM, Kagan D, Tran CA, Zeitvogel A, Starzinski-Powitz A, Nataraja S, et al. Tumor necrosis factor-alpha regulates inflammatory and mesenchymal responses via mitogen-activated protein kinase kinase, p38, and nuclear factor kappaB in human endometriotic epithelial cells. Mol Pharmacol. 2008;73:1394–404.

  40. 40.

    Xiao J, Gong Y, Chen Y, Yu D, Wang X, Zhang X, et al. IL-6 promotes epithelial-to-mesenchymal transition of human peritoneal mesothelial cells possibly through the JAK2/STAT3 signaling pathway. Am J Physiol Renal Physiol. 2017;313:F310–8.

  41. 41.

    Lee SO, Yang X, Duan S, Tsai Y, Strojny LR, Keng P, et al. IL-6 promotes growth and epithelial-mesenchymal transition of CD133+ cells of non-small cell lung cancer. Oncotarget. 2016;7:6626–38.

  42. 42.

    Li S, Lu J, Chen Y, Xiong N, Li L, Zhang J, et al. MCP-1-induced ERK/GSK-3β/Snail signaling facilitates the epithelial-mesenchymal transition and promotes the migration of MCF-7 human breast carcinoma cells. Cell Mol Immunol. 2017;14:621–30.

  43. 43.

    Lee CH, Wu CL, Shiau AL. Toll-like receptor 4 signaling promotes tumor growth. J Immunother. 2010;33:73–82.

  44. 44.

    Sujitha S, Dinesh P, Rasool M. Berberine modulates ASK1 signaling mediated through TLR4/TRAF2 via upregulation of miR-23a. Toxicol Appl Pharmacol. 2018;359:34–46.

  45. 45.

    Rahimifard M, Maqbool F, Moeini-Nodeh S, Niaz K, Abdollahi M, Braidy N, et al. Targeting the TLR4 signaling pathway by polyphenols: A novel therapeutic strategy for neuroinflammation. Ageing Res Rev. 2017;36:11–9.

  46. 46.

    Joh EH, Gu W, Kim DH. Echinocystic acid ameliorates lung inflammation in mice and alveolar macrophages by inhibiting the binding of LPS to TLR4 in NF-kB and MAPK pathways. Biochem Pharmacol. 2012;84:40.

  47. 47.

    Zeng KW, Yu Q, Liao LX, Song FJ, Lv HN, Jiang Y, et al. Anti-neuroinflammatory effect of MC13, a novel coumarin compound from condiment murraya, through inhibiting lipopolysaccharide-Induced TRAF6-TAK1-NF-κB, P38/ERK MAPKS and Jak2-Stat1/Stat3 pathways. J Cell Biochem. 2015;116:1286–99.

  48. 48.

    Wang Y, Shan X, Dai Y, Jiang L, Chen G, Zhang Y, et al. Curcumin analog L48H37 prevents lipopolysaccharide-induced TLR4 signaling pathway activation and sepsis via targeting MD2. J Pharmacol Exp Ther. 2015;353:539–50.

  49. 49.

    Kim SY, Koo JE, Seo YJ, Tyagi N, Jeong E, Choi J, et al. Suppression of Toll-like receptor 4 activation by caffeic acid phenethyl ester is mediated by interference of LPS binding to MD2. Br J Pharmacol. 2013;168:1933–45.

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Acknowledgements

This work was supported by Ramalingaswami re-entry fellowship (BT/RLF/Re-entry/36/2013). S. R. is the recipient of Ramalingaswami re-entry fellowship from the Department of Biotechnology (DBT), Government of India. This study was also supported in part by the Department of Science and Technology (DST; Award No: YSS/2014/000125) (to S. R.), Government of India. The first author (A. S.) gratefully acknowledges the support of Indian Council of Medical Research (ICMR), New Delhi, India for the award of ICMR‐Senior Research Fellowship (SRF; Award No: 45/03/2018‐BMS/PHA/OL). The infrastructure of Department of Biotechnology, Anna University, BIT-campus is supported by the Department of Science and Technology-Fund for Improvement of S and T Infrastructure in Universities and Higher Educational Institutions (DST-FIST).

Author information

SR has conceptualized, designed the experiments, acquired financial support, supervised the study, and wrote the manuscript; AS performed most of the experiments; DP and NR involved in the acquisition of data; AS, DP and SR analysed data and interpreted the results; AK and SKM participated in cytokine analysis; LL and KM participated in computational analysis; RB participated in flow cytometry analysis; RB, SKM and RP contributed reagents/materials/analysis tools.

Correspondence to Subbiah Rajasekaran.

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Sivanantham, A., Pattarayan, D., Rajasekar, N. et al. Tannic acid prevents macrophage-induced pro-fibrotic response in lung epithelial cells via suppressing TLR4-mediated macrophage polarization. Inflamm. Res. 68, 1011–1024 (2019). https://doi.org/10.1007/s00011-019-01282-4

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Keywords

  • EMT
  • LPS
  • M1 macrophages
  • Mesenchymal cells
  • Tannic acid
  • TLR4