Ischemia and Reperfusion of Liver Induces eNOS and iNOS Expression: Effects of a NO Donor and NOS Inhibitor

Hen I Lin；David Wang；Fur-Jiang Leu；Chao-Fuh Chen；Hsing I Chen

ischemia-reperfusion liver injury ； NOS ； NO donor ； NOS inhibitor

The Chinese Journal of Physiology

47卷3期（2004 / 09 / 30）

121 - 127

英文

The aim of this study was to investigate the role of nitric oxide (NO) in hepatic ischemiareperfusion (I/R) injury in rats. Immunohistochemistry was used to examine the protein expression of endothelial and inducible nitric oxide synthases (eNOS, iNOS) and nitrotyrosine after hR challenges to the liver, and blood levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactic dehydrogenase (LDH), hydroxyl radical and NO were measured before ischemia and after reperfusion. Ischemia was induced by occlusion of the common hepatic artery and portal vein for 40 mm, followed by reperfusion for 90 min. Reperfusion of the liver induced a significant increase in the blood concentrations of AST, ALT, LDH (n=8; P＜0.001), hydroxyl radical (n=8; P＜0.001) and NO (n=8; P＜0.01). The eNOS, iNOS, nitrotyrosine, SOD1 and SOD2 protein expression was also found to increase significantly after reperfusion (n=3). Administration of the NOS inhibitor N(superscript ω)-nitro-L-arginine methyl ester (L-NAME) (n=8) had a protective effect on the I/R-related injury, but the NO donor L-arginine (L-Arg) (n=8) potentiated the damage caused by I/R. These results suggest that reperfusion of the liver induces expression of NOS, which is related to the elevation of blood NO. The increase in hydroxyl radical concentration was accompanied by an increase in antioxidant enzyme expression (SOD1 and SOD2), and an increase in nitrotyrosine expression was also observed, reflecting the increased production of NO and oxygen radicals. We concluded from the protective effect of L-NAME and the potentiation by L-Arg that NOS expression and increases in NO and hydroxyl radical production have deleterious effects on the response to I/R in the liver.

1. Alexander, B.(1998).The role of nitric oxide in hepatic metabolism.Nutrition,14,376-390.
2. Bilzer, M.,Jaeschke, H.,Vollmar, A.M.,Paumgartner, G.,Gerbes, A.L.(1999).Prevention of Kopffer cell-induced oxidant injury in rat liver by atrial natriuretic peptide.Am. J. Physiol.,276,1137-1144.
3. Bowes, J.,Thiemermann, C.(1998).Effects of inhibition of the activity of poly (ADP-ribose) synthase on the liver injury caused by ischemia-reperfusion: a comparison with radical scavengers.Br. J. Pharmacol.,124,1254-1260.
4. Chen, C.F.,Wang, D.,Hwang, C.P.,Liu, H.W.,Wei, J.,Lee, R.P.,Chen, H.I.(2001).The protective effect of niacinamide on ischemia-reperfusion-induced liver injury.J. Blamed. Sci.,8,446-452.
5. Cottart, C.H.,Do, L.,Blanc, M.C.,Vaubourdolle, M.,Descamps, G.,Durand, D.,Galen, F.X.,Clot, J.P.(1999).Hepatoprotective effect of endogenous nitric oxide during ischemia-reperfusion in the rat.Hepatology,29,809-813.
6. Crow, J.P.,Beckman, J.S.(1995).Reactions between nitric oxide, superoxide and peroxynitrite: footprints of peroxynitrite in vivo.Ady. Pharmacol.,34,17-43.
7. Furchgott, R.F.,Zawadzki, J.V.(1980).The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine.Nature,288,373-376.
8. Hierholzer, C.,Harbrecht, B.,Menezes, J.M.,Kane, J.,MacMicking, J.,Nathan, C.F.,Peitzman, A.B.,Billiar, T.R.,Tweardy, D.J.(1998).Essential role of induced nitric oxide in the initiation of the inflammatory response after hemorrhagic shock.J. Exp. Med.,187,917-928.
9. Hsu, C.M.,Wang, J.S.,Liu, C.H.,Chen, L.W.(2002).Kupffer cells protect liver from ischemia-reperfusion injury by an inducible nitric oxide synthase-dependent mechanism.Shock,17,280-285.
10. Isobe, M.,Katsuramaki, T.,Kimura, H.,Nagayama, M.,Mauuno, T.,Yagihashi, A.,Hirata, K.(2000).Role of inducible nitric oxide synthase on hepatic ischemia and reperfusion injury.Transplant. Proc.,32,1650-1652.
11. Kawachi, S.,Hines, I.N.,Laronx, F. S.,Hoffman, J.,Bharwani, S.,Gray, L.,Leffer, D.,Grisham, M.B.(2000).Nitric oxide synthase and postischemic liver injury.Biochem. Biophys. Res. Commun.,276,851-854.
12. Kimura, H.,Katsuramaki, T.,Isobe, M.,Nagayama, M.,Meguro, M.,Kokita, K.,Nui, A.,Hirata, K.(2003).Role of inducible nitric oxide synthase in pig liver transplantation.J. Surg. Res.,111,28-37.
13. Koken, T.,Inal, M.(1999).The effect of nitric oxide on ischemia-reperfusion injury in rat liver.Clin. Chim. Acta.,288,55-62.
14. Kubes, P.(1993).Ischemia-reperfusion in feline small intestine: a role for nitric oxide.Am. J. Physiol.,264,143-149.
15. Lee, V.G.,Johnson, M.L.,Baust, J.,Laobach, V.E.,Watkins, S.C.,Billiar, T.R.(2001).The roles of iNOS in liver ischemia-reperfusion injury.Shock,16,355-360.
16. Liu, P.,Xu, B.,Spokas, E.,Lai, P.S.,Wong, P.Y.(2000).Role of endogenous nitric oxide in TNF-alpha and IL-1beta generation in hepatic ischemia-repefusion.Shock,13,217-223.
17. Liu, T.H.,Robinson, E.K.,Helmer, K.S.,West, S.D.,Castaneda, A.A.,Chang, L.,Mercer, D.W.(2002).Does upregulation of inducible nitric oxide synthase play a role in hepatic injury?.Shock,18,549-554.
18. Meguro, M.,Katsuramaki, T.,Nagayama, M.,Kimura, H.,Isobe, M.,Kimura, Y.,Matsuno, T.,Nui, A.,Hirata, K.(2002).A novel inhibitor of inducible nitric oxide synthase (ONO-1714) prevents critical warm ischemia-reperfusion injury in the pig liver.Trans plantation,73,1439-1446.
19. Mercer, D.W.,Castaneda, A.A.,Denning, J.W.,Chang, L.,Rossell, D.H.(1998).Effects of endotoxin on gastric injury from luminal irritants in rats: potential roles of nitric oxide.Am. J. Physiol.,275,449-459.
20. Nakamura, K.,Jenaga, K.,Yokozawa, T.,Fujitsuka, N.,Oura, H.(1991).Production of methyl guanidine from creatinine via cretol by active oxygen species: analysis of the catabolism in vitro.Nephron,58,42-46.
21. Serracino-Inglott, F.,Virlos, I. T.,Habib, N.A.,Williamson, R.C.,Mathie, RT.(2003).Differential nitric oxide synthase expression during hepatic ischemia-reperfusion.Am. J. Surg.,185,589-595.
22. Szabo, C.,Billiar, T.R.(1999).Novel roles of nitric oxide in hemorrhagic shock.Shock,12,1-9.
23. Wahl, S.M.,McCartney-Francis, N.,Chan, J.,Dionne, R.,Ta, L.,Orenstein, J.M.(2003).Nitric oxide in experimental joint inflammation. Benefit or detriment?.Cells Tissues Organs,174,26-33.
24. Walsh, T.R.,Rao, P.N.,Makowka, L.,Starzl, T.E.(1990).Lipid peroxidation is a nonparenchymal cell event with reperfusion after prolonged liver ischemia.J. Surg. Res.,49,18-22.
25. Wang, W.,Xu, Z.,Lin, L.,Wang, W.,Li, D.,Wang, Z.(2000).Protective effect of L-arginine on liver during ischemia-reperfusion injury.Zhonghua Gan Zang Bing Za Zhi,8,370-372.

1. Wang, Yan-ling,Liu, Jian-pei,Liu, De-zhao,Huang, Pin-jie,Hei, Zi-qing,Gan, Xiao-liang(2014).Effects of Anti-Histamine Treatment on Liver Injury Triggered by Small Intestinal Ischemia Reperfusion in Rats.中國生理學雜誌,57(5),271-278.
2. Wu, Ming-Shun,Pan, Wen-Harn,Owaga, Eddy,Cheng, Shu-Yu,Chang, Jung-Su,Bai, Chyi-Huey(2014).Relationship between Hemoglobin Levels and Risk for Suspected Non-Alcoholic Fatty Liver in Taiwanese Adults.中國生理學雜誌,57(5),286-294.
3. Yang, Victoria-Hui,Shen, Amy-Ru,Liou, Teh-Ling,Lee, Chia-Wei,Hwu, Chii-Min,Huang, Zih-Ling,Chien, Eileen Jea,Chien, Chau-Heng(2006).Inducible Nitric Oxide Synthase Expression and Plasma Bilirubin Changes in Rats under Intermittent Hypoxia Treatment.Chinese Journal of Physiology,49(5),275-280.