Ameliorative effects of silymarin on HCl-induced acute lung injury in rats; role of the Nrf-2/HO-1 pathway

Document Type: Original Article

Authors

1 Department of Medical Biochemistry, Faculty of Medicine, Minia University, 61511 Minia, Egypt

2 Department of Pathology, Faculty of Medicine, Minia University, 61511 Minia, Egypt.

3 Research Building, Urology & Nephrology Center, Mansoura University, 35516 Mansoura, Egypt.

4 Department of Pharmacology, Faculty of Medicine, Minia University, 61511 Minia, Egypt

10.22038/ijbms.2019.14069

Abstract

Objective(s): Aspiration is a common cause of acute lung injury (ALI), which lacks an effective treatment. Inflammation and oxidative stress play key roles in ALI development. Silymarin is an active extract of Silybum marianum plant seeds (milk thistle). Silymarin has potent anti-inflammatory and antioxidant effects; however its role in aspiration induced ALI has not been investigated. The aim of this study is to investigate the role of silymarin in the treatment of hydrochloric acid (HCl) aspiration induced ALI and explores its mechanisms of action.
Materials and Methods: The study included three groups of rats: Control non-treated group, ALI group (intra-tracheal HCl injected), and silymarin treated ALI group. White blood cells (WBCs) with differential count, oxidative stress parameters, B-cell lymphoma 2 (Bcl-2), transforming growth factor-beta (TGF-β), cyclooxygenase 2 (COX-2), nuclear factor erythroid 2-related factor-2 (Nrf-2), and heme oxygenase-1 (HO-1) were investigated. Lung tissue histopathology and immunohistochemical expression of survivin and proliferating cell nuclear antigen (PCNA) were also examined.
Results: The results of the study showed that HCL caused histopathological changes in ALI with leukocytopenia and increased oxidative stress biomarkers. It increased TGF-β, up-regulated mRNA expression of COX-2, Nrf-2, and HO-1 and increased survivin and PCNA but decreased Bcl-2. Silymarin ameliorated the histopathological lung injury with further up-regulation of Nrf-2 and HO-1 mRNA and decreased the inflammatory and fibrotic parameters together with up-regulation of the anti-apoptotic and the proliferation parameters.
Conclusion: The protective effect of silymarin against ALI is mediated by Nrf-2/HO-1 pathway with subsequent antioxidant, anti-inflammatory, antiapoptotic, and proliferating activities.

Keywords


1. Qing R, Huang Z, Tang Y, Xiang Q, Yang F. Cordycepin alleviates lipopolysaccharide-induced acute lung injury via Nrf2/HO-1 pathway. Int Immunopharmacol 2018; 60:18-25.
2.     Qian L, Zhao Y, Guo L, Li S, Wu X. Activating transcription factor 3 (ATF3) protects against lipopolysaccharide-induced acute lung injury via inhibiting the expression of TL1A. J Cell Physiol 2017; 232:3727-3734.
3.     Ashbaugh DG, Bigelow DB, Petty TL, Levine BE. Acute respiratory distress in adults. Lancet 1967; 2:319-323.
4.     Tsushima K, King LS, Aggarwal NR, De Gorordo A, D’Alessio FR, Kubo K. Acute lung injury review. Internal medicine 2009; 48:621-630.
5.     Reiss LK, Uhlig U, Uhlig S. Models and mechanisms of acute lung injury caused by direct insults. Eur J Cell Biol 2012; 91:590-601.
6.     Terasaki Y, Terasaki M, Urushiyama H, Nagasaka S, Takahashi M, Kunugi S, et al. Role of survivin in acute lung injury: epithelial cells of mice and humans. Lab Invest 2013; 93:1147-1163.
7.     Zhao F, Shi D, Li T, Li L, Zhao M. Silymarin attenuates paraquat-induced lung injury via Nrf2-mediated pathway in vivo and in vitro. Clin Exp Pharmacol Physiol 2015; 42:988-998.
8.     Davidson BA, Vethanayagam RR, Grimm MJ, Mullan BA, Raghavendran K, Blackwell TS, et al. NADPH oxidase and Nrf2 regulate gastric aspiration-induced inflammation and acute lung injury. J Immunol 2013; 190:1714-1724.
9.     Li H, Zhou X, Tan H, Hu Y, Zhang L, Liu S, et al. Neutrophil extracellular traps contribute to the pathogenesis of acid-aspiration-induced ALI/ARDS. Oncotarget 2017; 9:1772-1784.
10. Puig F, Herrero R, Guillamat-Prats R, Gomez MN, Tijero J, Chimenti L, et al. A new experimental model of acid- and endotoxin-induced acute lung injury in rats. Am J Physiol Lung Cell Mol Physiol 2016; 311:229-237.
11. Lee JM, Li J, Johnson DA, Stein TD, Kraft AD, Calkins MJ, et al. Nrf2, a multi-organ protector?. FASEB J 2005; 19:1061-1066.
12. Ungvari Z, Bagi Z, Feher A, Recchia FA, Sonntag WE, Pearson K, et al. Resveratrol confers endothelial protection via activation of the antioxidant transcription factor Nrf2. Am J Physiol Heart Circ Physiol 2010; 299:18-24.
13. Surai PF. silymarin as a natural antioxidant: An overview of the current evidence and perspectives. Antioxidants 2015; 4:204-247.
14. Katiyar SK. Silymarin and skin cancer prevention: anti-inflammatory, antioxidant and immunomodulatory effects (review). Int J Oncol 2005; 26:169-176.
15. Jia R, Cao L, Du J, Xu P, Jeney G, Yin G. The protective effect of silymarin on the carbon tetrachloride (CCl4)-induced liver injury in common carp (Cyprinus carpio). In Vitro Cell Dev Biol Anim 2013; 49:155-161.
16. Demir M, Amanvermez R, Kamali Polat A, Karabicak I, Cinar H, Kesicioglu T, et al. The effect of silymarin on mesenteric ischemia-reperfusion injury. Med Princ Pract 2014; 23:140-144.
17. Neha, Jaggi AS, Singh N. Silymarin and its role in chronic diseases. Adv Exp Med Biol 2016; 929:25-44.
18. Zhu Z, Sun G. Silymarin mitigates lung impairments in a rat model of acute respiratory distress syndrome. Inflammopharmacol 2018; 26:747-754.
19. Liu HM, Guo YN. Effect of captopril on serum TNF-alpha level in acute lung injury rats induced by HCL. Asian Pac J Trop Med 2014; 7:905-908.
20. Wang L, Huang QH, Li YX, Huang YF, Xie JH, Xu LQ, et al. Protective effects of silymarin on triptolide-induced acute hepatotoxicity in rats. Mol Med Rep 2018; 17:789-800.
21. Atef  Y, El-Fayoumi HM, Abdel-Mottaleb Y, Mahmoud MF. Quercetin and tin protoporphyrin attenuate hepatic ischemia reperfusion injury: role of HO-1. Naunyn-Schmiedeberg’s Arch Pharmacol 2017; 390:871-881.
22. Kostopanagiotou G, Avgerinos E, Costopanagiotou C, Arkadopoulos N, Andreadou I, Diamantopoulou K, et al. Acute lung injury in a rat model of intestinal ischemia-reperfusion: the potential time depended role of phospholipases A(2). J Surg Res 2008; 147:108-116.
23. Setzer F, Schmidt B, Hueter L, Schwarzkopf K, Sanger J, Schreiber T. Characterization of the seven-day course of pulmonary response following unilateral lung acid injury in rats. PloS One 2018; 13:198440-198459.
24. Folkesson HG, Matthay MA, Hebert CA, Broaddus VC. Acid aspiration-induced lung injury in rabbits is mediated by interleukin-8-dependent mechanisms. J Clin Invest 1995; 96:107-116.
25. Booth CM, Matukas LM, Tomlinson GA, Rachlis AR, Rose DB, Dwosh HA, et al. Clinical features and short-term outcomes of 144 patients with SARS in the greater Toronto area. JAMA 2003; 289:2801-2809.
26. Jalali SM, Najafzadeh H, Mousavi S. Comparative effect of silymarin and d-penicillamine on lead induced hemotoxicity and oxidative stress in Rat. Iran J Toxicol 2017; 1:12-18
27. Gust R, Kozlowski JK, Stephenson AH, Schuster DP. Role of cyclooxygenase-2 in oleic acid-induced acute lung injury. Am J Respir Crit Care Med 1999; 160:1165-1170.
28. Terao Y, Nakamura T, Morooka H, Sumikawa K. Effect of cyclooxygenase-2 inhibitor pretreatment on gas exchange after hydrochloric acid aspiration in rats. J Anesth 2005; 19:257-259.
29. Robertson JA, Sauer D, Gold JA, Nonas SA. The role of cyclooxygenase-2 in mechanical ventilation-induced lung injury. Am J Respir Cell Mol Biol 2012; 47:387-394.
30. El-Lakkany NM, Hammam OA, El-Maadawy WH, Badawy AA, Ain-Shoka AA, Ebeid FA. Anti-inflammatory/anti-fibrotic effects of the hepatoprotective silymarin and the schistosomicide praziquantel against Schistosoma mansoni-induced liver fibrosis. Parasite Vector 2012; 5:9-23.
31. Ramakrishnan G, Elinos-Baez CM, Jagan S, Augustine TA, Kamaraj S, Anandakumar P, et al. Silymarin downregulates COX-2 expression and attenuates hyperlipidemia during NDEA-induced rat hepatocellular carcinoma. Mol Cell Biochem 2008; 313:53-61.
32. Sharifi R, Pasalar P, Kamalinejad M, Dehpour AR, Tavangar SM, Paknejad M, et al. The effect of silymarin (Silybum marianum) on human skin fibroblasts in an in vitro wound healing model. Pharm Biol 2013; 51:298-303.
33. Toklu HZ, Tunali Akbay T, Velioglu-Ogunc A, Ercan F, Gedik N, Keyer-Uysal M, et al. Silymarin, the antioxidant component of Silybum marianum, prevents sepsis-induced acute lung and brain injury. J Surg Res 2008; 145:214-222.
34. Guzel A, Kanter M, Aksu B, Basaran UN, Yalcin O, Guzel A, et al. Preventive effects of curcumin on different aspiration material-induced lung injury in rats. Pediatr Surg Int 2009; 25:83-92.
35. Yilmaz MZ, Guzel A, Torun AC, Okuyucu A, Salis O, Karli R, et al. The therapeutic effects of anti-oxidant and anti-inflammatory drug quercetin on aspiration-induced lung injury in rats. J Mol Histol 2014; 45:195-203.
36. Kurutas EB. The importance of antioxidants which play the role in cellular response against oxidative/nitrosative stress: current state. Nutr J 2016; 15:71-93.
37. Abdel-Gaber SA, Mohammed RK, Refaie MMM. Mechanism mediating the protective effect of diacerein in ischemia-reperfusion-induced testicular injury in rats. Life Sci 2018; 209:57-62.
38. Kong M, Mao J, Luo B, Qin Z. Role of transcriptional factor Nrf2 in the acute lung injury of mice. Int J Clin Exp Pathol 2015; 8:10929-10934.
39. Hsu HH, Ko WJ, Hsu JY, Chen JS, Lee YC, Lai IR, et al. Simvastatin ameliorates established pulmonary hypertension through a heme oxygenase-1 dependent pathway in rats. Respir Res 2009; 10:32-45.
40. Otterbein LE, Soares MP, Yamashita K, Bach FH. Heme oxygenase-1: unleashing the protective properties of heme. Trends Immunol 2003; 24:449-455.
41. Leffler CW, Nasjletti A, Yu C, Johnson RA, Fedinec AL, Walker N. Carbon monoxide and cerebral microvascular tone in newborn pigs. Am J Physiol 1999; 276:1641-1646.
42. Kawamura T, Wakabayashi N, Shigemura N, Huang CS, Masutani K, Tanaka Y, et al. Hydrogen gas reduces hyperoxic lung injury via the Nrf2 pathway in vivo. Am J Physiol Lung Cell Mol Physiol 2013; 304:646-656.
43. Joo Choi R, Cheng MS, Shik Kim Y. Desoxyrhapontigenin up-regulates Nrf2-mediated heme oxygenase-1 expression in macrophages and inflammatory lung injury. Redox Biol 2014; 2:504-512.
44. Osburn WO, Wakabayashi N, Misra V, Nilles T, Biswal S, Trush MA, et al. Nrf2 regulates an adaptive response protecting against oxidative damage following diquat-mediated formation of superoxide anion. Arch Biochem Biophys 2006; 454:7-15.
45. Yang CC, Fang JY, Hong TL, Wang TC, Zhou YE, Lin TC. Potential antioxidant properties and hepatoprotective effects of an aqueous extract formula derived from three Chinese medicinal herbs against CCl(4)-induced liver injury in rats. Int Immunopharmacol 2013; 15:106-113.
46. Toklu HZ, Tunali-Akbay T, Erkanli G, Yuksel M, Ercan F, Sener G. Silymarin, the antioxidant component of Silybum marianum, protects against burn-induced oxidative skin injury. Burns 2007; 33:908-916.
47. Raskovic A, Stilinovic N, Kolarovic J, Vasovic V, Vukmirovic S, Mikov M. The protective effects of silymarin against doxorubicin-induced cardiotoxicity and hepatotoxicity in rats. Mol 2011; 16:8601-8613.
48. Roubalova L, Dinkova-Kostova AT, Biedermann D, Kren V, Ulrichova J, Vrba J. Flavonolignan 2,3-dehydrosilydianin activates Nrf2 and upregulates NAD(P)H:quinone oxidoreductase 1 in Hepa1c1c7 cells. Fitoterapia 2017; 119:115-120.
49. Turkseven S, Kruger A, Mingone CJ, Kaminski P, Inaba M, Rodella LF, et al. Antioxidant mechanism of heme oxygenase-1 involves an increase in superoxide dismutase and catalase in experimental diabetes. Am J Physiol Heart Circ Physiol 2005; 289:701-707.
50. Buccellato LJ, Tso M, Akinci OI, Chandel NS, Budinger GR. Reactive oxygen species are required for hyperoxia-induced Bax activation and cell death in alveolar epithelial cells. J Biol Chem 2004; 279:6753-6760.
51. An J, Park SH, Ko IG, Jin JJ, Hwang L, Ji ES, et al. Polydeoxyribonucleotide ameliorates lipopolysaccharide-induced lung injury by inhibiting apoptotic cell death in rats. Int J Mol Sci 2017;18:1847-1861.
52. Bao XC, Fang YQ, You P, Zhang S, Ma J. Protective role of peroxisome proliferator-activated receptor beta/delta in acute lung injury induced by prolonged hyperbaric hyperoxia in rats. Respir Physiol Neurobiol 2014; 199:9-18.
53. Raza SS, Khan MM, Ashafaq M, Ahmad A, Khuwaja G, Khan A, et al. Silymarin protects neurons from oxidative stress associated damages in focal cerebral ischemia: a behavioral, biochemical and immunohistological study in Wistar rats. J Neurol Sci 2011; 309:45-54.
54. Chtourou Y, Garoui E, Boudawara T, Zeghal N. Therapeutic efficacy of silymarin from milk thistle in reducing manganese-induced hepatic damage and apoptosis in rats. Hum Exp Toxicol 2013; 32:70-81.
55. El-Sheikh AA, Morsy MA, Hamouda AH. Protective mechanisms of thymoquinone on methotrexate-induced intestinal toxicity in rats. Pharmacogn Mag 2016; 12:76-81.
56. Ganeshan K, Johnston LK, Bryce PJ. TGF-beta1 limits the onset of innate lung inflammation by promoting mast cell-derived IL-6. J Immunol 2013;190:5731-5738.
57. Broekelmann TJ, Limper AH, Colby TV, McDonald JA. Transforming growth factor beta 1 is present at sites of extracellular matrix gene expression in human pulmonary fibrosis. Proc Natl Acad Sci U S A 1991; 88:6642-6646.
58. Clichici S, Olteanu D, Nagy AL, Oros A, Filip A, Mircea PA. Silymarin inhibits the progression of fibrosis in the early stages of liver injury in CCl(4)-treated rats. J Med Food 2015; 18:290-298.
59. Kantah MK, Kobayashi R, Sollano J, Naito Y, Solimene U, Jains S, et al. Hepatoprotective activity of a phytotherapeutic formula on thioacetamide--induced liver fibrosis model. Acta Biomed 2011; 82:82-89.
60. Amenomori  S, Terasaki Y, Terasaki M, Urusiyama H, Takahashi M, Kunugi S, et al. The increased expression of survivin on lipopolysaccharaide (LPS)-induced acute lung injury (ALI) in mice. Am J Respir Crit Care Med 2011;183:2098-2099.
61. Mishra A, Doyle NA, Martin WJc. Bleomycin-mediated pulmonary toxicity: evidence for a p53-mediated response. Am J Respir Cell Mol Biol 2000; 22:543-549.
62. Hastings RH, Asirvatham A, Quintana R, Sandoval R, Dutta R, Burton DW, et al. Parathyroid hormone-related protein-(38-64) regulates lung cell proliferation after silica injury. Am J Physiol Lung Cell Mol Physiol 2002; 283:12-21.
63. Cherniack RM, Colby TV, Flint A, Thurlbeck WM, Waldron J, Ackerson L, et al. Quantitative assessment of lung pathology in idiopathic pulmonary fibrosis. Am Rev Respir Dis 1991; 144:892-900.
64. Mishra A, Liu JY, Brody AR, Morris GF. Inhaled asbestos fibers induce p53 expression in the rat lung. Am J Respir Cell Mol Biol 1997; 16:479-485.
65. Malaviya R, Venosa A, Hall L, Gow AJ, Sinko PJ, Laskin JD, et al. Attenuation of acute nitrogen mustard-induced lung injury, inflammation and fibrogenesis by a nitric oxide synthase inhibitor. Toxicol Appl Pharmacol 2012; 265:279-291.
66. He Q, Kim J, Sharma RP. Silymarin protects against liver damage in BALB/c mice exposed to fumonisin B1 despite increasing accumulation of free sphingoid bases. Toxicol Sci 2004; 80:335-342.