Amitriptyline attenuates bleomycin-induced pulmonary fibrosis: modulation of the expression of NF-κβ, iNOS, and Nrf2

  • Mai A. ZaafanEmail author
  • Ahmed R. Haridy
  • Amr M. Abdelhamid
Original Article


Amitriptyline is a tricyclic antidepressant that was suggested to have antifibrotic potential. The current study aimed to investigate the modulatory effects of amitriptyline on bleomycin-induced pulmonary fibrosis in rats. Rats were randomly assigned into 4 groups: normal control, bleomycin control, amitriptyline+bleomycin, and amitriptyline only treated group. Lung injury was evaluated through the histological examination and immunohistochemical detection of α-smooth muscle actin (α-SMA) in lung tissue, in addition to the biochemical assessment of pulmonary contents of hydroxyproline and transforming growth factor beta-1 (TGF-β1). In addition, the following parameters were investigated for studying the possible mechanisms of amitriptyline antifibrotic effect: inducible nitric oxide synthase (iNOS), nuclear factor-κβ (NF-κβ), tumor necrosis factor-alpha (TNF-α), serpine-1, p53, nuclear factor erythroid 2-related factor 2 (Nrf2), lipid peroxides, and reduced glutathione (GSH). Amitriptyline exhibited potent antifibrotic effect that was reflected upon the histopathological examination and through its ability to suppress all the fibrotic parameters. Amitriptyline successfully suppressed the expression of NF-κβ, Nrf2, iNOS, and p53 in lung tissues besides the inhibition of other oxidative stress and inflammatory mediators. Amitriptyline could be a promising treatment to pulmonary fibrosis. Amitriptyline not only prevents the depression and its drawbacks in patients suffering from pulmonary fibrosis but also it can suppress fibrosis through variable mechanisms mainly via inhibition of NF-κβ/TNF-α/TGF-β pathway in addition to inhibition of Nrf2 and iNOS expression.


Amitriptyline Pulmonary fibrosis Nuclear factor-κβ Nrf2 iNOS 



The authors are very grateful to Dr. Adel M. Bakeer, Professor of Pathology, Faculty of Veterinary Medicine, Cairo University, for examining and interpreting histopathologic aspects of this study. In addition, the authors are very grateful to our dear students; Marina Rafeal, Aghaby Youhana and Mahmoud Ismail who helped in the practical part of the study.

Author contribution

MA and AR conceived and designed the research. MA and AM conducted experiments. AM and AR analyzed data. MA wrote the manuscript. All authors read and approved the manuscript.

Compliance with ethical standards

The study was done with compliance to the ethics standards and approval from the ethics committee of the October University for Modern Sciences and Arts, Egypt.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Achar E, Maciel TT, Collares CF, Teixeira VP, Schor N (2009) Amitriptyline attenuates interstitial inflammation and ameliorates the progression of renal fibrosis. Kidney Int 75(6):596–604CrossRefGoogle Scholar
  2. Artaud-Macari E, Goven D, Brayer S, Hamimi A, Besnard V, Marchal-Somme J, Ali ZE, Crestani B, Kerdine-Römer S, Boutten A, Bonay M (2013) Nuclear factor erythroid 2-related factor 2 nuclear translocation induces myofibroblastic dedifferentiation in idiopathic pulmonary fibrosis. Antioxid Redox Signal 18(1):66–79CrossRefGoogle Scholar
  3. Banchroft P, Stevens A, Turner DR (1996) Theory and practice of histological techniques, 4th edn. Churchil Livingstone, LondonGoogle Scholar
  4. Chen JY, Newcomb B, Zhou C, Pondick JV, Ghoshal S, York SR et al (2017) Tricyclic antidepressants promote ceramide accumulation to regulate collagen production in human hepatic stellate cells. Sci Rep 7(1):1–13Google Scholar
  5. Cheresh P, Kim SJ, Tulasiram S, Kamp DW (2013) Oxidative stress and pulmonary fibrosis. Biochim Biophys Acta 7(40):5Google Scholar
  6. Guo C, Atochina-Vasserman E, Abramova H, George B, Manoj V, Scott P, Gow A (2016) Role of NOS2 in pulmonary injury and repair in response to bleomycin. Free Radic Biol Med 91(1):293–301CrossRefGoogle Scholar
  7. He X, Wang L, Szklarz G, Bi Y, Ma Q (2012) Resveratrol inhibits paraquat-induced oxidative stress and fibrogenic response by activating the nuclear factor erythroid 2-related factor 2 pathway. J Pharmacol Exp Ther 342(1):81–90CrossRefGoogle Scholar
  8. Jiang C, Liu G, Luckhardt T, Antony V, Zhou Y, Carter AB, Thannickal VJ, Liu RM (2017) Serpine 1 induces alveolar type II cell senescence through activating p53-p21-Rb pathway in fibrotic lung disease. Aging Cell 16(5):1114–1124CrossRefGoogle Scholar
  9. Kikuchi N, Ishii Y, Morishima Y, Yageta Y, Haraguchi N, Itoh K et al (2010) Nrf2 protects against pulmonary fibrosis by regulating the lung oxidant level and Th1/Th2 balance. Respir Res 11(31):1465–9921Google Scholar
  10. Lekkerkerker AN, Aarbiou J, van Es T, Janssen RA (2012) Cellular players in lung fibrosis. Curr Pharm Des 18(27):4093–4102CrossRefGoogle Scholar
  11. Li C, Yu Y, Li W, Liu B, Jiao X, Song X, Lv C, Qin S (2017a) Phycocyanin attenuates pulmonary fibrosis via the TLR2-MyD88-NF-kappaB signaling pathway. Sci Rep 7(1):5843–5855CrossRefGoogle Scholar
  12. Li M, Li Y, Li J, Zhao P, Bai Y, Feng S et al (2017b) Long-term effects of TCM Yangqing Kangxian formula on bleomycin-induced pulmonary fibrosis in rats via regulating nuclear factor-kappaB signaling. Evid Based Complement Alternat Med 2017(1):1–16Google Scholar
  13. Liu Y, Huang G, Mo B, Wang C (2017) Artesunate ameliorates lung fibrosis via inhibiting the Notch signaling pathway. Exp Ther Med 14(1):561–566CrossRefGoogle Scholar
  14. Liu M, Xu H, Zhang L, Zhang C, Yang L, Ma E, Liu L, Li Y (2018) Salvianolic acid B inhibits myofibroblast transdifferentiation in experimental pulmonary fibrosis via the up-regulation of Nrf2. Biochem Biophys Res Commun 495(1):325–331CrossRefGoogle Scholar
  15. Luzina IG, Todd NW, Sundararajan S, Atamas SP (2015) The cytokines of pulmonary fibrosis: much learned, much more to learn. Cytokine 74(1):88–100CrossRefGoogle Scholar
  16. Marudamuthu AS, Shetty SK, Bhandary YP, Karandashova S, Thompson M, Sathish V, Florova G, Hogan TB, Pabelick CM, Prakash YS, Tsukasaki Y, Fu J, Ikebe M, Idell S, Shetty S (2015) Plasminogen activator inhibitor-1 suppresses profibrotic responses in fibroblasts from fibrotic lungs. J Biol Chem 290(15):9428–9441CrossRefGoogle Scholar
  17. Matsuda T, Taniguchi H, Ando M, Kondoh Y, Kimura T, Kataoka K, Nishimura K, Nishiyama O, Sakamoto K, Hasegawa Y (2017) Depression is significantly associated with the health status in patients with idiopathic pulmonary fibrosis. Intern Med 56(13):1637–1644CrossRefGoogle Scholar
  18. Qiu W, Wu M, Liu S, Chen B, Pan C, Yang M, Wang KJ (2017) Suppressive immunoregulatory effects of three antidepressants via inhibition of the nuclear factor-kappaB activation assessed using primary macrophages of carp (Cyprinus carpio). Toxicol Appl Pharmacol 322(1):1–8CrossRefGoogle Scholar
  19. Tian SL, Yang Y, Liu XL, Xu QB (2018) Emodin attenuates bleomycin-induced pulmonary fibrosis via anti-inflammatory and anti-oxidative activities in rats. Med Sci Monit 24(1):1–10CrossRefGoogle Scholar
  20. Tukey JW (1951) Reminder sheets for “Discussion of paper on multiple comparisons by Henry Scheffe”. In the collected work of John W. Tukey VIII. Multiple comparisons. Chapman and Hall, New YorkGoogle Scholar
  21. Verma R, Brahmankar M, Kushwah L, Suresh B (2013) Evaluating the inhibitory potential of sulindac against the bleomycin-induced pulmonary fibrosis in Wistar rats. Environ Toxicol Pharmacol 36(3):769–778CrossRefGoogle Scholar
  22. Wuyts WA, Agostini C, Antoniou KM, Bouros D, Chambers RC, Cottin V, Egan JJ, Lambrecht BN, Lories R, Parfrey H, Prasse A, Robalo-Cordeiro C, Verbeken E, Verschakelen JA, Wells AU, Verleden GM (2013) The pathogenesis of pulmonary fibrosis: a moving target. Eur Respir J 41(5):1207–1218CrossRefGoogle Scholar
  23. Yao C, Li A, Gao W, Pallua N, Steffens G (2011) Improving the angiogenic potential of collagen matrices by covalent incorporation of Astragalus polysaccharides. Int J Burns Trauma 1(1):17–26PubMedPubMedCentralGoogle Scholar
  24. Zaafan MA, Zaki HF, El-Brairy AI, Kenawy SA (2016) Pyrrolidinedithiocarbamate attenuates bleomycin-induced pulmonary fibrosis in rats: modulation of oxidative stress, fibrosis, and inflammatory parameters. Exp Lung Res 42(8–10):408–416CrossRefGoogle Scholar
  25. Zhang Z, Qu J, Zheng C, Zhang P, Zhou W, Cui W, Mo X, Li L, Xu L, Gao J (2018) Nrf2 antioxidant pathway suppresses Numb-mediated epithelial-mesenchymal transition during pulmonary fibrosis. Cell Death Dis 9(2):83–94CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Pharmacology and Toxicology Department, Faculty of PharmacyOctober University for Modern Sciences and Arts (MSA)6th of OctoberEgypt
  2. 2.Biochemistry Department, Faculty of PharmacyOctober University for Modern Sciences and Arts (MSA)6th of OctoberEgypt

Personalised recommendations