BACKGROUND & AIMS: Ischemia-reperfusion (I/R) is a major mechanism of liver injury following hepatic surgery or transplantation. Despite numerous reports on the role of oxidative/nitrosative stress and mitochondrial dysfunction in hepatic I/R injury, the proteins that are oxidatively modified during I/R damage are poorly characterized. This study was aimed at investigating the oxidatively modified proteins underlying the mechanism for mitochondrial dysfunction in hepatic I/R injury. We also studied the effects of a superoxide dismutase mimetic/peroxynitrite scavenger metalloporphyrin (MnTMPyP) on oxidatively modified proteins and their functions. METHODS: The oxidized and/or S-nitrosylated mitochondrial proteins from I/R-injured mouse livers with or without MnTMPyP pretreatment were labeled with biotin-N-maleimide, purified with streptavidin-agarose, and resolved by 2-dimensional gel electrophoresis. The identities of the oxidatively modified proteins were determined using mass spectrometric analysis. Liver histopathology, serum transaminase levels, nitrosative stress markers, and activities of oxidatively modified mitochondrial proteins were measured. RESULTS: Comparative 2-dimensional gel analysis revealed markedly increased numbers of oxidized and S-nitrosylated mitochondrial proteins following hepatic I/R injury. Many key mitochondrial enzymes involved in cellular defense, fat metabolism, energy supply, and chaperones were identified as being oxidatively modified proteins. Pretreatment with MnTMPyP attenuated the I/R-induced increased serum transaminase levels, histologic damage, increased inducible nitric oxide synthase expression, and S-nitrosylation and/or nitration of various key mitochondrial proteins. MnTMPyP pretreatment also restored I/R-induced suppressed activities of mitochondrial aldehyde dehydrogenase, 3-ketoacyl-CoA thiolases, and adenosine triphosphate synthase. CONCLUSIONS: These results suggest that increased nitrosative stress is critically important in promoting S-nitrosylation and nitration of various mitochondrial proteins, leading to mitochondrial dysfunction with decreased energy supply and increased hepatic injury.
BACKGROUND & AIMS:Ischemia-reperfusion (I/R) is a major mechanism of liver injury following hepatic surgery or transplantation. Despite numerous reports on the role of oxidative/nitrosative stress and mitochondrial dysfunction in hepatic I/R injury, the proteins that are oxidatively modified during I/R damage are poorly characterized. This study was aimed at investigating the oxidatively modified proteins underlying the mechanism for mitochondrial dysfunction in hepatic I/R injury. We also studied the effects of a superoxide dismutase mimetic/peroxynitrite scavenger metalloporphyrin (MnTMPyP) on oxidatively modified proteins and their functions. METHODS: The oxidized and/or S-nitrosylated mitochondrial proteins from I/R-injured mouse livers with or without MnTMPyP pretreatment were labeled with biotin-N-maleimide, purified with streptavidin-agarose, and resolved by 2-dimensional gel electrophoresis. The identities of the oxidatively modified proteins were determined using mass spectrometric analysis. Liver histopathology, serum transaminase levels, nitrosative stress markers, and activities of oxidatively modified mitochondrial proteins were measured. RESULTS: Comparative 2-dimensional gel analysis revealed markedly increased numbers of oxidized and S-nitrosylated mitochondrial proteins following hepatic I/R injury. Many key mitochondrial enzymes involved in cellular defense, fat metabolism, energy supply, and chaperones were identified as being oxidatively modified proteins. Pretreatment with MnTMPyP attenuated the I/R-induced increased serum transaminase levels, histologic damage, increased inducible nitric oxide synthase expression, and S-nitrosylation and/or nitration of various key mitochondrial proteins. MnTMPyP pretreatment also restored I/R-induced suppressed activities of mitochondrial aldehyde dehydrogenase, 3-ketoacyl-CoA thiolases, and adenosine triphosphate synthase. CONCLUSIONS: These results suggest that increased nitrosative stress is critically important in promoting S-nitrosylation and nitration of various mitochondrial proteins, leading to mitochondrial dysfunction with decreased energy supply and increased hepatic injury.
Authors: C A Bradham; R F Stachlewitz; W Gao; T Qian; S Jayadev; G Jenkins; Y Hannun; J J Lemasters; R G Thurman; D A Brenner Journal: Hepatology Date: 1997-05 Impact factor: 17.425
Authors: Agnieszka A Kendrick; Mahua Choudhury; Shaikh M Rahman; Carrie E McCurdy; Marisa Friederich; Johan L K Van Hove; Peter A Watson; Nicholas Birdsey; Jianjun Bao; David Gius; Michael N Sack; Enxuan Jing; C Ronald Kahn; Jacob E Friedman; Karen R Jonscher Journal: Biochem J Date: 2011-02-01 Impact factor: 3.857
Authors: Partha Mukhopadhyay; Mohanraj Rajesh; Béla Horváth; Sándor Bátkai; Ogyi Park; Galin Tanchian; Rachel Y Gao; Vivek Patel; David A Wink; Lucas Liaudet; György Haskó; Raphael Mechoulam; Pál Pacher Journal: Free Radic Biol Med Date: 2011-03-11 Impact factor: 7.376
Authors: Xi Yang; James Greenhaw; Qiang Shi; Dean W Roberts; Jack A Hinson; Levan Muskhelishvili; Kelly Davis; William F Salminen Journal: J Pharmacol Exp Ther Date: 2012-10-23 Impact factor: 4.030
Authors: Mohamed A Abdelmegeed; Sehwan Jang; Atrayee Banerjee; James P Hardwick; Byoung-Joon Song Journal: Free Radic Biol Med Date: 2013-02-27 Impact factor: 7.376
Authors: Ignazio Grattagliano; Catia V Diogo; Maria Mastrodonato; Ornella de Bari; Michele Persichella; David Q H Wang; Adriana Liquori; Domenico Ferri; Maria Rosaria Carratù; Paulo J Oliveira; Piero Portincasa Journal: World J Gastroenterol Date: 2013-05-28 Impact factor: 5.742