Pulmonary MnSOD is nitrated following hepatic ischemia-reperfusion.
BACKGROUND
Ischemia-reperfusion (I/R) of remote organs is a common cause of lung injury. We observed that lung injury after partial hepatic I/R in mice coincides with the appearance of 3-nitrotyrosine (NT) in the lung tissue, a marker of peroxynitrite involvement and oxidant stress. Peroxynitrite can cause mitochondrial dysfunction by inactivation of manganese superoxide dismutase (MnSOD), the major antioxidant enzyme in mitochondria. Our aims were to examine whether pulmonary MnSOD is a target of nitration following hepatic I/R and whether nitrated MnSOD (N-MnSOD) correlates with acute lung injury.


METHODS
Five 20-25-g male C57BL/6 mice underwent laparotomy, and atraumatic occlusion of the portal and arterial blood supply to the upper three lobes of the liver for 90 min. This warm ischemic period was followed by 4 h of reperfusion, and the animals were then euthanized. Lung injury was assessed by LDH and protein levels in bronchoalveolar lavage (BAL) fluid. Pulmonary MnSOD activity in pulmonary homogenates was measured by the cytochrome c reduction method. The presence of N-MnSOD was determined by immunoprecipitation (IP) and Western Blot analysis. Controls (N = 5) underwent sham operation.


RESULTS
Elevated plasma transaminases confirmed hepatic injury. Lung injury was demonstrated by elevation in BAL protein and LDH levels (495.7 (48.4) versus 644.9 (37.3) [p < 0.05] and 56.5 (11.8) versus 345.2 (80) [p < 0.01], respectively). Immunoprecipitation and Western blot demonstrated N-MnSOD in the lung tissue of I/R animals but not controls. MnSOD activity decreased following I/R (8.1 (0.7) versus 10.8 (0.3) [p < 0.05]).


CONCLUSIONS
Pulmonary MnSOD is both nitrated and inactivated following hepatic I/R and is associated with acute lung injury. These findings suggest that MnSOD incapacitance may contribute to I/R-induced lung injury and provide a therapeutic target in attenuating multisystem injury following hepatic I/R.
Gray
Keith D
KD
Department of Surgery at Vanderbilt University Medical Center, Nashville, Tennessee, USA.
MacMillan-Crow
Lee-Ann
LA
Simovic
Misho O
MO
Stain
Steven C
SC
May
Addison K
AK
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001707:Biopsy, Needle; D004195:Disease Models, Animal; D007150:Immunohistochemistry; D007511:Ischemia / Q000150:complications / Q000628:therapy*; D008099:Liver / Q000098:blood supply*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018384:Oxidative Stress; D015424:Reperfusion / Q000009:adverse effects* / Q000379:methods; D015427:Reperfusion Injury / Q000473:pathology; D012128:Respiratory Distress Syndrome / Q000201:enzymology* / Q000473:pathology; D012680:Sensitivity and Specificity; D012720:Severity of Illness Index; D013482:Superoxide Dismutase / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism*
D015415:Biomarkers; C002744:3-nitrotyrosine; D014443:Tyrosine; D013482:Superoxide Dismutase