Nigella sativa attenuates bleomycin-induced pulmonary fibrosis in rats by inhibition of inflammation, fibrosis, and inducible nitric oxide synthase

Year 2015, Volume: 32 Issue: 3, 121 - 125, 17.11.2015

Mustafa Erboga , Zeynep Fidanol Erboga Yeliz Bozdemır Donmez Cevat Aktas , Mehmet Kanter

Abstract

The present study investigated the anti-oxidant, anti-inflammatory and anti-fibrotic potential of Nigella sativa (NS) against bleomycin (BLC)-induced lung injury and fibrosis, an experimental animal model. A total of 18 male Sprague-Dawley rats were divided into three groups: Control, BLC, BLC+NS; each group consist of 6 animals. Pulmonary fibrosis was induced by a single intratracheal instillation of 2.5 mg/kg BLC. BLC+NS group rats were intragastric administered daily 400 mg/kg of NS from day 1 to 28. At the end of the study, lung tissues were removed for histopathological and immunohistochemical investigation. Lung tissues were stained with hematoxylin and eosin and Masson’s Trichrome for histological evaluation. Moreover, the inducible nitric oxide synthase (iNOS) expression in the lung tissues was determined by immunohistochemical staining. BLC-induced histological changes including lung inflammation and lung fibrosis were significantly detected compared to the control group. NS treatment significantly ameliorated the BLC mediated histological changes and reduced the inflammatory cell infiltrate in lung tissues. NS significantly blocked collagen accumulations with parallel reduction in the fibrosis score. In addition, NS also markedly decreased the positive staining of iNOS in lung tissues. Our study provides evidence that NS significantly ameliorated BLM-induced pulmonary fibrosis in rats via the inhibition of inflammation score, fibrosis score, and iNOS expression. Therefore, NS may be a potential therapeutic reagent for the treatment of lung fibrosis.

Keywords

Bleomycin, Inducible nitric oxide synthase, Nigella sativa, Pulmonary fibrosis, Rat

References

• Adamson, I.Y., 1976. Pulmonary toxicity of bleomycin. Environ. Health. Perspect. 16, 119-125.
• Ahmad, A., Husain, A., Mujeeb, M., Khan, S.A., Najmi, A.K., Siddique, N.A., 2013. Damanhouri Z.A, Anwar F.A review on therapeutic potential of Nigella sativa: A miracle herb. Asian. Pac. J. Trop. Biomed. 3, 337-352. doi: 10.1016/S2221-1691(13)60075-1.
• Akhtar, M., Maikiyo, A.M., Najmi, A.K., Khanam, R., Mujeeb, M., Aqil, M., 2013. Neuroprotective effects of chloroform and petroleum ether extracts of Nigella sativa seeds in stroke model of rat. J. Pharm. Bioallied. Sci. 5, 119-125. doi: 10.4103/0975-7406.111825.
• Alemi, M., Sabouni, F., Sanjarian, F., Haghbeen, K., Ansari, S., 2013. Anti-inflammatory effect of seeds and callus of Nigella sativa L. Extracts on mix glial cells with regard to their thymoquinone content. AAPS. Pharm. Sci. Tech. 14, 160-167. doi: 10.1208/s12249-012-9899-8.
• American Thoracic Society (ATS), 2000. Idiopathic pulmonary fibrosis: Diagnosis and treatment. International consensus statement. ATS, and the European Respiratory Society (ERS). Am. J. Respir. Crit. Care. Med. 161, 646-664.
• Barnes, P.J., Hansel, T.T., 2004. Prospects for new drugs for chronic obstructive pulmonary disease. Lancet. 364, 985-996.
• Bourgou, S., Pichette, A., Marzouk, B., Legault, J., 2012. Antioxidant, antiinflammatory, anticancer and antibacterial activities of extracts from Nigella sativa (Black Cumin) plant parts. J. Food. Biochem. 36, 539-546.
• Chaudhary, N.I., Schnapp, A., Park, J.E., 2006. Pharmacologic differentiation of inflammation and fibrosis in the rat bleomycin model. Am. J. Respir. Crit. Care. Med. 173, 769-776.
• Chen, C.Y., Peng, W.H., Wu, L.C., Wu, C.C., Hsu, S.L., 2010. Luteolin ameliorates experimental lung fibrosis both in vivo and in vitro: Implications for therapy of lung fibrosis. J. Agric. Food. Chem. 58, 11653-11661. doi: 10.1021/jf1031668.
• Claussen, C.A., Long, E.C., 1999. Nucleic Acid recognition by metal complexes of bleomycin. Chem. Rev. 99, 2797-2816.
• Di Paola, R., Talero, E., Galuppo, M., Mazzon, E., Bramanti, P., Motilva, V., Cuzzocrea, S., 2011. Adrenomedullin in inflammatory process associated with experimental pulmonary fibrosis. Respir. Res. 12, 41. doi: 10.1186/1465-9921-12-41.
• Erden, E.Ş., Kırkıl, G., Deveci, F., İlhan, N., Çobanoğlu, B., Turgut, T., Muz, M.H., 2008. Ratlarda bleomisin ile oluşturulan akciğer fibrozisinde erdosteinin inflamasyon ve fibrozis üzerine etkileri. Tüberküloz ve Toraks Derg. 56, 127-138.
• Ermis, H., Parlakpinar, H., Gulbas, G., Vardi, N., Polat, A., Cetin, A., Kilic, T., Aytemur, Z.A., 2013. Protective effect of dexpanthenol on bleomycin-induced pulmonary fibrosis in rats. Naunyn. Schmiedebergs. Arch. Pharmacol. 386, 1103-1110. doi: 10.1007/s00210-013-0908-6.
• Galuppo, M., Esposito, E., Mazzon, E., Di Paola, R., Paterniti, I., Impellizzeri, D., Cuzzocrea, S., 2011. MEK inhibition suppresses the development of lung fibrosis in the bleomycin model. Naunyn. Schmiedebergs. Arch. Pharmacol. 384, 21-37. doi: 10.1007/s00210-011- 0637-7.
• Gao, Y., Lu, J., Zhang, Y., Chen, Y., Gu, Z., Jiang, X., 2013. Baicalein attenuates bleomycin-induced pulmonary fibrosis in rats through inhibition of miR-21. Pulm. Pharmacol. Ther. 26, 649-654. doi: 10.1016/j.pupt.2013.03.006.
• Hagiwara, S., Ishii, Y., Kitamura, S., 2000. Aerolized administration of N-acetylcysteine attenuates lung fibrosis induced by bleomicin in mice. Am. J. Respir. Crit. Care. Med. 162, 225-231.
• Hobbs, A.J., Higgs, A., Moncada, S., 1999. Inhibition of nitric oxide synthase as a potential therapeutic target. Annu. Rev. Pharmacol. Toxicol. 39, 191-220.
• Hsu, S.M., Raine, L., Fanger, H., 1981. Use of avidin biotinperoxidase complex (ABC) in immunperoxidase techniques: A comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29, 577-580.
• Hubner, R.H., Gitter, W., El Mokhtari, N.E., Mathiak, M., Both, M., Bolte, H., Freitag-Wolf, S., Bewig, B., 2008. Standardized quantification of pulmonary fibrosis in histological samples. Biotechniques. 44, 507-511, 514-517. doi: 10.2144/000112729.
• Iraz, M., Erdogan, H., Kotuk, M., Yağmurca, M., Kilic, T., Ermis, H., Fadillioğlu, E., Yildirim, Z., 2006. Ginkgo biloba inhibits bleomycin- induced lung fibrosis in rats. Pharmacol. Res. 53, 310-316.
• Kalayarasan, S., Sriram, N., Sudhandiran, G., 2008. Diallyl sulfide attenuates bleomycin-induced pulmonary fibrosis: Critical role of iNOS, NF- kappaB, TNF-alpha and IL-1beta. Life. Sci. 82, 1142-1153. doi: 10.1016/j.lfs.2008.03.018.
• Karimfar, M.H., Rostami, S., Haghani, K., Bakhtiyari, S., Noori-Zadeh, A., 2015. Melatonın allevıates bleomycın-induced pulmonary fıbrosıs in mıce. J. Biol. Regul. Homeost. Agents. 29, 327-334.
• Kilic, T., Ciftci, O., Cetin, A., Kahraman, H., 2014. Preventive effect of chrysin on bleomycin-induced lung fibrosis in rats. Inflammation. 37, 2116-2124. doi: 10.1007/s10753-014-9946-6.
• Kilic, T., Parlakpinar, H., Taslidere, E., Yildiz, S., Polat, A., Vardi, N., Colak, C., Ermis, H., 2015. Protective and therapeutic effect of apocynin on bleomycin-induced lung fibrosis in rats. Inflammation. 38, 1166-1180. doi: 10.1007/s10753-014-0081-1.
• Lakari, E., Soini, Y., Saily, M., Koistinen, P., Paakko, P., Kinnula, V.L., 2002. Inducible nitric oxide synthase, but not xanthine oxidase, is highly expressed in interstitial pneumonias and granulomatous diseases of human lung. Am. J. Clin. Pathol. 117, 132-142.
• Larki, A., Hemmati, A.A., Arzi, A., Borujerdnia, M.G., Es maeilzadeh, S., Zad Karami, M.R., 2013. Regulatory effect of caffeic acid phenethyl ester on type I collagen and interferon-gamma in bleomycin-induced pulmonary fibrosis in rat. Res. Pharm. Sci. 8, 243-252.
• Li, S., Yang, X., Li, W., Li, J., Su, X., Chen, L., Yan, G., 2012. N-acetylcysteine downregulation of lysyl oxidase activity alleviating bleomycin- induced pulmonary fibrosis in rats. Respiration. 84, 509-517. doi: 10.1159/000340041.
• Mouratis, M.A., Aidinis, V., 2011. Modeling pulmonary fibrosis with bleomycin. Curr. Opin. Pulm. Med. 17, 355-361. doi: 10.1097/ MCP.0b013e328349ac2b.
• Moeller, A., Ask, K., Warburton, D., Gauldie, J., Kolb, M., 2008. The bleomycin animal model: A useful tool to investigate treatment options for idiopathic pulmonary fibrosis? Int. J. Biochem. Cell. Biol. 40, 362-382.
• Salem, M.L., 2005. Immunomodulatory and immunotherapeutic properties of the Nigella sativa L. seed. Int. Immunopharmacol. 5, 1749-1770.
• Salem, E.M., Yar, T., Bamosa, A.O., Al-Quorain, A., Yasawy, M.I., Alsulaiman, R.M., Randhawa, M.A., 2010. Comparative study of Nigella sativa and triple therapy in eradication of Helicobacter pylori in patients with non-ulcer dyspepsia. Saudi. J. Gastroenterol. 16, 207-214. doi: 10.4103/1319-3767.65201.
• Sultan, M., Butt, M.S., Karim, R., Iqbal, S.Z., Ahmad, S., Zia-Ul-Haq, M., Aliberti, L., Ahmad, A.N., De Feo, V., 2014. Effect of Nigella sativa fixed and essential oils on antioxidant status, hepatic enzymes, and immunity in streptozotocin induced diabetes mellitus. BMC. Complement. Altern. Med. 14, 193. doi: 10.1186/1472-6882-14-193.
• Swamy, S.M., Tan, B.K., 2000. Cytotoxic and immunopotentiating effects of ethanolic extract of Nigella sativa L. seeds. J. Ethnopharmacol. 70, 1-7.
• Usuki, K.F.Y., 1995. Evolution of three patterns of intra-alveolar fibrosis produced by bleomycin in rats. Pathol. Int. 45, 552-564.
• Verma, R., Kushwah, L., Gohel, D., Patel, M., Marvania, T., Balakrishnan, S., 2013. Evaluating the ameliorative potential of Quercetin against the Bleomycin-induced pulmonary fibrosis in Wistar Rats. Pulm. Med. 2013, 921724. doi: 10.1155/2013/921724.