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Thymoquinone mitigate ischemia-reperfusion-induced liver injury in rats: a pivotal role of nitric oxide signaling pathway

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Abstract

Oxidative and nitrosative stress-induced endothelial cell damage play an essential role in the pathogenesis of hepatic ischemia-reperfusion (IR) injury. IR is associated with reduced eNOS expression and exacerbated by superimposed stress. NOSTRIN induces intracellular endothelial nitric oxide synthase (eNOS) translocation and inducible nitric oxide synthase (iNOS) increases nitric oxide (NO) production. Our aim was to assess hepatic expression of iNOS, eNOS, and NOSTRIN in IR with or without N-acetylcysteine (NAC) or thymoquinone (TQ) pretreatment and to compare their hepatoprotective effects. Surgical induction of IR was performed by occlusion of hepatic pedicle for 30 min with mini-clamp and reperfused for 30 min. The effects of TQ (20 mg/kg/day) or NAC (300 mg/kg/day) administered orally for 10 days were evaluated by serum ALT and AST, oxidative stress parameters, NO production, and histopathological analysis. Also, localization and expression of iNOS, eNOS, and NOSTRIN were assessed by immunofluorescence. TQ or NAC pretreatment significantly decreased elevated serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and myeloperoxidase (MPO) activities, malondialdehyde (MDA) level, and NO production. In addition, they restored the depleted GSH content and alleviated histopathological changes. Furthermore, they up-regulated eNOS and down-regulated iNOS and NOSTRIN expressions. TQ exerts its hepatoprotective effect, at least in part, by nitric oxide signaling pathway through modulation of iNOS, eNOS, and NOSTRIN expressions as well as suppression of oxidative stress.

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References

  • Abd-Elbaset M, Arafa E-SA, El Sherbiny GA, Abdel-Bakky MS, Elgendy ANA (2015) Quercetin modulates iNOS, eNOS and NOSTRIN expressions and attenuates oxidative stress in warm hepatic ischemia-reperfusion injury in rats Beni-Suef University. Journal of Basic and Applied Sciences 4:246–255

    Article  Google Scholar 

  • Al-Majed AA, Al-Omar FA, Nagi MN (2006) Neuroprotective effects of thymoquinone against transient forebrain ischemia in the rat hippocampus. Eur J Pharmacol 543:40–47

    Article  CAS  PubMed  Google Scholar 

  • Ara C et al (2005) Protective effect of resveratrol against oxidative stress in cholestasis. J Surg Res 127:112–117. doi:10.1016/j.jss.2005.01.024

    Article  CAS  PubMed  Google Scholar 

  • Attri S, Rana SV, Vaiphei K, Sodhi CP, Katyal R, Goel RC, Nain CK, Singh K (2000) Isoniazid- and rifampicin-induced oxidative hepatic injury--protection by N-acetylcysteine. Hum Exp Toxicol 19(9):517–522

  • Bergamini S, Rota C, Canali R, Staffieri M, Daneri F, Bini A, Giovannini F, Tomasi A, Iannone A (2001) N-acetylcysteine inhibits in vivo nitric oxide production by inducible nitric oxide synthase. Nitric Oxide 5(4):349–60

  • Beutler E, Duron O, Kelly B (1963) Improved method for the determination of blood glutathione. J Lab Clin Med 61:882–888

  • El-Abhar H, Abdallah D, Saleh S (2003) Gastroprotective activity of < i > Nigella sativa</i > oil and its constituent, thymoquinone, against gastric mucosal injury induced by ischaemia/reperfusion in rats. J Ethnopharmacol 84:251–258

    Article  CAS  PubMed  Google Scholar 

  • El-Agamy DS, Nader MA (2012) Attenuation of oxidative stress-induced vascular endothelial dysfunction by thymoquinone. Exp Biol Med 237:1032–1038

    Article  CAS  Google Scholar 

  • El-Ghany R, Sharaf N, Kassem L, Mahran L, Heikal O (2009) Thymoquinone triggers anti-apoptotic signaling targeting death ligand and apoptotic regulators in a model of hepatic ischemia reperfusion injury. Drug discoveries & therapeutics 3:296–306

    Google Scholar 

  • El-Mahmoudy A, Matsuyama H, Borgan MA, Shimizu Y, El-Sayed MG, Minamoto N, Takewaki T (2002) Thymoquinone suppresses expression of inducible nitric oxide synthase in rat macrophages. Int Immunopharmacol 2:1603–1611

  • Gali-Muhtasib H, Roessner A, Schneider-Stock R (2006) Thymoquinone: a promising anti-cancer drug from natural sources. Int J Biochem Cell Biol 38:1249–1253

    Article  CAS  PubMed  Google Scholar 

  • Hemalatha P, Reddy AG, Reddy YR, Shivakumar P (2013) Evaluation of protective effect of N-acetyl cysteine on arsenic-induced hepatotoxicity. J Nat Sci Biol Med 4:393–395. doi:10.4103/0976-9668.116986

    Article  PubMed  PubMed Central  Google Scholar 

  • Icking A, Matt S, Opitz N, Wiesenthal A, Müller-Esterl W, Schilling K (2005) NOSTRIN functions as a homotrimeric adaptor protein facilitating internalization of eNOS. J Cell Sci 118:5059–5069

    Article  CAS  PubMed  Google Scholar 

  • Jaeschke H (2003) Molecular mechanisms of hepatic ischemia-reperfusion injury and preconditioning. Am J Physiol Gastrointest Liver Physiol 284:G15–G26. doi:10.1152/ajpgi.00342.2002

    Article  CAS  PubMed  Google Scholar 

  • Jones LM, Bancroft JD, Gamble M (2008) Theory and practice of histological techniques. 6th ed. London: Churchill Livingstone, p 744

  • Krawisz JE, Sharon P, Stenson WF (1984) Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Assessment of inflammation in rat and hamster models Gastroenterology 87(6):1344-50

  • Mihara M, Uchiyama M (1978) Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem 86(1):271-8

  • Miranda KM, Espey MG, Wink DA (2001) A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide 5(1):62-71

  • Montalvo-Jave EE, Escalante-Tattersfield T, Ortega-Salgado JA, Piña E, Geller DA (2008) Factors in the pathophysiology of the liver ischemia-reperfusion injury. J Surg Res 147:153–159

    Article  CAS  PubMed  Google Scholar 

  • Mookerjee RP, Wiesenthal A, Icking A, Hodges SJ, Davies NA, Schilling K, Sen S, Williams R, Novelli M, Müller-Esterl W, Jalan R (2007) Increased gene and protein expression of the novel eNOS regulatory protein NOSTRIN and a variant in alcoholic hepatitis. Gastroenterology 132(7):2533-2541

  • NAGI MN, MANSOUR MA (2000) Protective effect of thymoquinone against doxorubicin–induced cardiotoxicity in rats: a possible mechanism of protection. Pharmacol Res 41:283–289

    Article  CAS  PubMed  Google Scholar 

  • Oguz A et al (2013) The effects of curcumin on the liver and remote organs after hepatic ischemia reperfusion injury formed with Pringle manoeuvre in rats. Eur Rev Med Pharmacol Sci 17:457–466

    CAS  PubMed  Google Scholar 

  • Prescott L, Illingworth R, Critchley J, Stewart M, Adam R, Proudfoot A (1979) Intravenous N-acetylcystine: the treatment of choice for paracetamol poisoning. BMJ 2:1097–1100

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schilling K, Opitz N, Wiesenthal A, Oess S, Tikkanen R, Muller-Esterl W, Icking A (2006) Translocation of endothelial nitric-oxide synthase involves a ternary complex with caveolin-1 and NOSTRIN. Mol Biol Cell 17:3870–3880. doi:10.1091/mbc.E05-08-0709

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sener G, Tosun O, Sehirli AO, Kaçmaz A, Arbak S, Ersoy Y, Ayanoğlu-Dülger G (2003) Melatonin and N-acetylcysteine have beneficial effects during hepatic ischemia and reperfusion. Life Sci 72(24):2707-2718

  • Sewerynek E, Reiter RJ, Melchiorri D, Ortiz GG, Lewinski A (1995) Oxidative damage in the liver induced by ischemia-reperfusion: protection by melatonin. Hepato-Gastroenterology 43:898–905

    Google Scholar 

  • Shah V, Kamath PS (2003) Nitric oxide in liver transplantation: pathobiology and clinical implications. Liver Transpl 9:1–11

    Article  PubMed  Google Scholar 

  • Thies J et al (1998) The efficacy of N-acetylcysteine as a hepatoprotective agent in liver transplantation. Transpl Int 11:S390–S392

    Article  PubMed  Google Scholar 

  • Thippeswamy T, McKay JS, Quinn JP, Morris R (2006) Nitric oxide, a biological double-faced janus—is this good or bad? Histol Histopathol 21(4):445–458

  • Trocha M, Merwid-Lad A, Szuba A, Chlebda E, Piesniewska M, Sozanski T, Szelag A (2010) Effect of simvastatin on nitric oxide synthases (eNOS, iNOS) and arginine and its derivatives (ADMA, SDMA) in ischemia/reperfusion injury in rat liver. Pharmacological reports : PR 62:343–351

    Article  CAS  PubMed  Google Scholar 

  • Xia Z, Liu M, Wu Y, Sharma V, Luo T, Ouyang J, McNeill JH (2006) N-acetylcysteine attenuates TNF-α-induced human vascular endothelial cell apoptosis and restores eNOS expression. Eur J Pharmacol 550:134–142

    Article  CAS  PubMed  Google Scholar 

  • Yildiz F et al (2008) Nigella sativa relieves the deleterious effects of ischemia reperfusion injury on liver. World journal of gastroenterology: WJG 14:5204

    Article  PubMed  PubMed Central  Google Scholar 

  • Zafarullah M, Li WQ, Sylvester J, Ahmad M (2003) Molecular mechanisms of N-acetylcysteine actions. Cell Mol Life Sci 60:6–20

    Article  CAS  PubMed  Google Scholar 

  • Zimmerman BJ, Granger DN (1994) Mechanisms of reperfusion injury. Am J Med Sci 307(4):284-292

  • Zimmermann K, Opitz N, Dedio J, Renne C, Muller-Esterl W, Oess S (2002) NOSTRIN: a protein modulating nitric oxide release and subcellular distribution of endothelial nitric oxide synthase. Proc Natl Acad Sci U S A 99:17167–17172. doi:10.1073/pnas.252345399

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, 62514, Egypt

    Mohamed Abd-Elbaset & El-Shaimaa A. Arafa

  2. Department of Pharmacology and Toxicology, College of Pharmacy and Health Sciences, Ajman University of Science and Technology, Ajman, United Arab Emirates

    El-Shaimaa A. Arafa

  3. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Kafr El-Sheikh University, Kafr El-Sheikh, Egypt

    Gamal A. El Sherbiny

  4. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt

    Mohamed S. Abdel-Bakky

  5. Department of Pharmacology, College of pharmacy, Al Jouf University, Al Jouf, Kingdom of Saudi Arabia

    Mohamed S. Abdel-Bakky

  6. Department of Pharmacology, Faculty of Veterinary medicine, Beni-Suef University, Beni-Suef, 62514, Egypt

    Abdel Nasser A.M. Elgendy

Authors
  1. Mohamed Abd-Elbaset
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  2. El-Shaimaa A. Arafa
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  3. Gamal A. El Sherbiny
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  4. Mohamed S. Abdel-Bakky
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  5. Abdel Nasser A.M. Elgendy
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Correspondence to El-Shaimaa A. Arafa.

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Abd-Elbaset, M., Arafa, ES.A., El Sherbiny, G.A. et al. Thymoquinone mitigate ischemia-reperfusion-induced liver injury in rats: a pivotal role of nitric oxide signaling pathway. Naunyn-Schmiedeberg's Arch Pharmacol 390, 69–76 (2017). https://doi.org/10.1007/s00210-016-1306-7

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  • DOI: https://doi.org/10.1007/s00210-016-1306-7

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