Abstract
We aimed to investigate the preventive and therapeutic effect of apocynin (APO) on bleomycin (BLC)-induced lung injury in rats. Rats were assigned into groups as follows: control group; APO group, 20 mg/kg APO was given intraperitoneal for 29 days; BLC-1 and BLC-2 groups, a single intratracheal injection of BLC (2.5 mg/kg); APO+BLC-preventive group, 20 mg/kg APO was administered 12 h before the intratracheal BLC injection and continued for 14 days; BLC+APO-treatment group, 20 mg/kg APO was given on the 14th day after the intratracheal BLC injection and continued to sacrifice. The BLC-1 group was sacrificed on the 14th day of BLC administration to validate BLC-induced lung inflammation and fibrosis on the 14th of study initiation. All other groups were sacrificed on the 29th day after BLC administration. The semiquantitative histopathological assessment, tissue levels of malondialdehyde (MDA), superoxide dismutase, catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), total antioxidant capacity, total oxidant status (TOS), and oxidative stress index (OSI) were measured. An addition to the serum myeloperoxidase (MPO), the cell count and cytokines (IL-1β, IL-6, and IL-8) of bronchoalveolar lavage (BAL) fluid were assayed. BLC-provoked histological changes were significantly detected compared to the control group. APO restored these histological damages in different quantity in the treatment and prevention groups. BLC caused a significant decrease in GSH, CAT, and GPX, which were accompanied with significantly the increased MDA, TOS levels, and OSI in the lung tissue concomitant with increased levels of the cellular account and proinflammatory cytokines in the BAL fluid. Otherwise, APO administration, both before and after BLC, reversed all biochemical markers and cytokine as well as histopathological changes induced by BLC. Interestingly, APO treatment reversed MPO activity in serum increased by BLC. In this study, both protective and therapeutic effects of APO against BLC-induced lung fibrosis were demonstrated for the first time.
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Raghu, G., H.R. Collard, J.J. Egan, et al. 2011. An official ATS/ERS/JRS/ALAT statement: idiopathic pulmonary fibrosis: evidence-based guidelines for diagnosis and management. American Journal of Respiratory and Critical Care Medicine 183: 788–824.
Gross, T.J., and G.W. Hunninghake. 2001. Idiopathic pulmonary fibrosis. The New England Journal of Medicine 345: 517–525.
Dedon, P.C., and I.H. Goldberg. 1992. Free-radical mechanisms involved in the formation of sequence-dependent bistranded DNA lesions by the antitumor antibiotics bleomycin, neocarzinostatin, and calicheamicin. Chemical Research in Toxicology 5: 311–332.
Cottin, V. 2013. The role of pirfenidone in the treatment of idiopathic pulmonary fibrosis. Respiratory Research 14: 5–9. doi:10.1186/1465-9921-14- S1-S5.
Mata, M., A. Ruíz, M. Cerdá, M. Martinez-Losa, et al. 2003. Oral N-acetylcysteine reduces bleomycin-induced lung damage and mucin Muc5ac expression in rats. The European Respiratory Journal 22: 900–905.
Sogut, S., H. Ozyurt, F. Armutcu, et al. 2004. Erdosteine prevents bleomycin-induced pulmonary fibrosis in rats. European Journal of Pharmacology 494: 213–220.
Yildirim, Z., Y. Turkoz, M. Kotuk, et al. 2004. Effects of aminoguanidine and antioxidant erdosteine on bleomycin-induced lung fibrosis in rats. Nitric Oxide 11: 156–165.
Kilic, T., H. Parlakpinar, A. Polat, et al. 2014. Protective and therapeutic effect of molsidomine on bleomycin-induced lung fibrosis in rats. Inflammation 37: 1167–1178. doi:10.1007/s10753-014-9841-1.
Yildirim, Z., M. Kotuk, H. Erdogan, et al. 2006. Preventive effect of melatonin on bleomycin-induced lung fibrosis in rats. Journal of Pineal Research 40: 27–33.
Iraz, M., H. Erdogan, M. Kotuk, et al. 2006. Ginkgo biloba inhibits bleomycin-induced lung fibrosis in rats. Pharmacological Research 53: 310–316.
Akgedik, R., S. Akgedik, H. Karamanli, et al. 2012. Effect of resveratrol on treatment of bleomycin-induced pulmonary fibrosis in rats. Inflammation 35: 1732–1741.
Ermis, H., H. Parlakpinar, G. Gulbas, et al. 2013. Protective effect of dexpanthenol on bleomycin-induced pulmonary fibrosis in rats. Naunyn-Schmiedeberg's Archives of Pharmacology 386: 1103–1110. doi:10.1007/s 00210-013-0908.
Kilic, T., O. Osman Ciftc, A. Cetin, et al. 2014. Protective effect of chrysin on bleomycin-induced lung fibrosis in rats. Inflammation. doi:10.1007/s10753-014-9946-6.
Stefanska, J, and R. Pawliczak. 2008. Apocynin: molecular aptitudes. Mediators of Inflammation 106507
Impellizzeri, D., E. Esposito, E. Mazzon, et al. 2011. Effect of apocynin, a NADPH oxidase inhibitor, on acute lung inflammation. Biochemical Pharmacology 81: 636–648.
De Caterina, R., and M. Massaro. 2005. Omega-3 fatty acids and the regulation of expression of endothelial proatherogenic and pro-inflammatory genes. The Journal of Membrane Biology 206: 103–116.
Taylor, N.E., P. Glocka, M. Liang, et al. 2006. NADPH oxidase in the renal medulla causes oxidative stress and contributes to salt-sensitive hypertension in Dahl S rats. Hypertension 47: 692–698.
Jin, L., R.A. Beswick, T. Yamamoto, et al. 2006. Increased reactive oxygen species contributes to kidney injury in mineralocorticoid hypertensive rats. Journal of Physiology and Pharmacology 57: 343–357.
Colak, C., and H. Parlakpınar. 2012. Hayvan Deneyleri: in vivo Denemelerin Bildirimi: ARRIVE Kılavuzu-Derleme. Journal Turgut Ozal Medical Center 19(2): 128–131.
Connell, B.J., M.C. Saleh, B.V. Khan, et al. 2011. Apocynin may limit total cell death following cerebral ischemia and reperfusion by enhancing apoptosis. Food and Chemical Toxicology 49: 3063–3069.
Uchiyama, M., and M. Mihara. 1978. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Analytical Biochemistry 34: 271–278.
Sun, Y., L. Oberley, and Y. Li. 1988. A simple method for clinical assay of superoxide dismutase. Clinical Chemistry 34: 497–500.
Aebi, H. 1974. Catalase. In Methods of enzymatic analysis, ed. H.U. Bergmeyer, 673–677. New York: Academic.
Paglia, D.E., and W.N. Valentine. 1967. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. Journal of Laboratory and Clinical Medicine 70: 158–170.
Ellman, G.L. 1959. Tissue sulfhydryl groups. Archives of Biochemistry and Biophysics 82: 70–77.
Bai, A., P. Hu, J. Chen, et al. 2007. Blockade of STAT3 by antisense oligonucleotide in TNBS-induced murine colitis. International Journal of Colorectal Disease 22: 625–635.
Erel, O. 2004. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clinical Biochemistry 37: 277–285.
Erel, O. 2005. A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry 38: 1103–1111.
Coward, W.R., G. Saini, and G. Jenkins. 2010. The pathogenesis of idiopathic pulmonary fibrosis. Therapeutic Advances in Respiratory Disease 4: 367–388.
Harari, S., and A. Caminati. 2010. IPF: new insight on pathogenesis and treatment. Allergy 65: 537–553.
Chaudhary, N.I., A. Schnapp, and J.E. Park. 2006. Pharmacologic differentiation of inflammation and fibrosis in the rat bleomycin model. American Journal of Respiratory and Critical Care Medicine 173: 769–776.
Mungunsukh, O., A.J. Griffin, Y.H. Lee, et al. 2010. Bleomycin induces the extrinsic apoptotic pathway in pulmonary endothelial cells. American Journal of Physiology - Lung Cellular and Molecular Physiology 298: 696–703.
Mouratis, M.A., and V. Aidinis. 2011. Modeling pulmonary fibrosis with bleomycin. Current Opinion in Pulmonary Medicine 17: 355–361.
Carre, P., and P. Leophonte. 1993. Cytokines and pulmonary fibroses. Revue des Maladies Respiratoires 10: 193–207.
Underwood, D.C., R.R. Osborn, S. Bochnowicz, et al. 2000. SB 239063, a p38 MAPK inhibitor, reduces neutrophilia, inflammatory cytokines, MMP-9, and fibrosis in lung. American Journal of Physiology - Lung Cellular and Molecular Physiology 279: L895–L902.
Gon, Y., S. Hashimoto, T. Nakayama, et al. 2000. N-Acetyl-l-cysteine inhibits bleomycin-induced interleukin-8 secretion by bronchial epithelial cells. Respirology 5: 309–313.
Sener, G., N. Topaloglu, A.O. Sehirli, et al. 2007. Resveratrol alleviates bleomycin-induced lung injury in rats. Pulmonary Pharmacology & Therapeutics 20: 642–649.
Chiang, C.H., C.H. Chuang, S.L. Liu, et al. 2011. Apocynin attenuates ventilator-induced lung injury in an isolated and perfused rat lung model. Intensive Care Medicine 37: 1360–1367.
Perg, D.W., T.M. Chang, J.Y. Wang, et al. 2013. Inflammatory role of AMP-activated protein kinase signaling in an experimental model of toxic smoke inhalation injury. Critical Care Medicine 41: 120–132.
Altintas, R., A. Polat, N. Nigar Vardi, et al. 2013. The protective effects of apocynin on kidney damage caused by renal ischemia/reperfusion. Journal of Endourology 27: 617–624.
Peters, E.A., J.T.N. Hiltermann, and J. Stolk. 2001. Effect of apocynin on ozone-induced airway hyperresponsiveness to methacholine in asthmatics. Free Radical Biology and Medicine 31: 1442–1447.
Ximenes, V.F., M.P. Kanegae, S.R. Rissato, et al. 2007. The oxidation of apocynin catalyzed by myeloperoxidase: proposal for NADPH oxidase inhibition. Archives of Biochemistry and Biophysics 457: 134–141.
Smit, H.F., B.H. Kroes, A.J.J. van den Berg, et al. 2000. Immunomodulatory and anti-inflammatory activity of Picrorhiza scrophulariiflora. Journal of Ethnopharmacology 73: 101–109.
Barbieri, S.S., V. Cavalca, S. Eligini, et al. 2004. Apocynin prevents cyclooxygenase 2 expression in human monocytes through NADPH oxidase and glutathione redox dependent mechanisms. Free Radical Biology and Medicine 37: 156–165.
Stefanska, J., A. Sarniak, A. Wlodarczyk, et al. 2012. Apocynin reduces reactive oxygen species concentrations in exhaled breath condensate in asthmatics. Experimental Lung Research 38: 90–99.
Hougee, S., A. Hartog, A. Sanders, et al. 2006. Oral administration of the NADPH-oxidase inhibitor apocynin partially restores diminished cartilage proteoglycan synthesis and reduces inflammation in mice. European Journal of Pharmacology 531: 264–269.
Tang, L.L., K. Ye, X.F. Yang, et al. 2007. Apocynin attenuates cerebral infarction after transient focal ischaemia in rats. Journal of International Medical Research 35: 517–522.
Xu, L., Y. Li, S. Wan, Y. Wang, and P. Yu. 2014. Protective effects of apocynin nitrone on acute lung injury induced by lipopolysaccharide in rats. International Immunopharmacology 20: 377–382.
Antao-Menes, A., E.A. Turpin, P.C. Post, et al. 2008. STAT-1 signaling in human lung fibroblast in induced vanadium pentoxide through an IFN-beta autocrine loop. Journal of Immunology 180: 4200–4207.
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The authors have no conflict of interest to declare. This work has been supported by the scientific research unit of Inonu University (Project number:2013/112 ).
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Kilic, T., Parlakpinar, H., Taslidere, E. et al. Protective and Therapeutic Effect of Apocynin on Bleomycin-Induced Lung Fibrosis in Rats. Inflammation 38, 1166–1180 (2015). https://doi.org/10.1007/s10753-014-0081-1
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DOI: https://doi.org/10.1007/s10753-014-0081-1