Amitriptyline attenuates bleomycin-induced pulmonary fibrosis: modulation of the expression of NF-κβ, iNOS, and Nrf2
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.
KeywordsAmitriptyline 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.
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.
- 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
- Banchroft P, Stevens A, Turner DR (1996) Theory and practice of histological techniques, 4th edn. Churchil Livingstone, LondonGoogle Scholar
- 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
- Cheresh P, Kim SJ, Tulasiram S, Kamp DW (2013) Oxidative stress and pulmonary fibrosis. Biochim Biophys Acta 7(40):5Google Scholar
- 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
- 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
- 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
- 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
- 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