title: Modulation of mitochondrial calcium management attenuates hepatic warm ischemia-reperfusion injury.
abstract: Hepatic warm ischemia and reperfusion (IR) injury occurs in many clinical situations and has an important link to subsequent hepatic failure. The pathogenesis of this injury involves numerous pathways, including mitochondrial-associated apoptosis. We studied the effect of mitochondrial calcium uptake inhibition on hepatic IR injury using the specific mitochondrial calcium uptake inhibitor, ruthenium red (RR). Rats were subjected to 1 hour of 70% warm hepatic ischemia following RR pretreatment or vehicle injection. Sham-operated animals served as controls. Analysis was performed at 15 minutes, 1 hour, 3 hours, or 6 hours after reperfusion. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) concentrations were determined. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining was performed to assess apoptosis, and hepatocellular necrosis was semiquantitated from hematoxylin and eosin-stained tissue sections. RR pretreatment significantly decreased both AST and ALT serum levels after 6 hours of reperfusion (AST: 1,556 +/- 181 U/L vs. 597 +/- 121 U/L, P = 0.005; ALT: 1,118 +/- 187 U/L vs. 294 +/- 39 U/L, P = 0.005). Apoptosis was observed within 15 minutes of reperfusion in vehicle-pretreated animals and peaked after 3 hours of reperfusion (98 +/- 21 cells/high-power field [hpf]). Apoptosis was inhibited at all time points by RR pretreatment. Histologic evidence of necrosis was not observed prior to 3 hours of reperfusion (23% +/- 4%), and maximal necrosis was observed after 6 hours of reperfusion (26% +/- 1% percent area). RR pretreatment significantly decreased the necrotic percent area at both the 3-hour and the 6-hour time points (4.2% +/- 2%; 3.7% +/- 1%, respectively). Hepatic IR injury resulted in both apoptotic and necrotic cell death, which were attenuated by RR pretreatment. In conclusion, these observations implicate mitochondrial calcium uptake in the pathogenesis of hepatic IR injury.
authors.affiliation: Department of Surgery, Division of Hepatobiliary Surgery and Liver Transplantation, Vanderbilt University Medical Center, Nashville, TN 37232-4753, USA.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Anderson
authors.forename: Janene
authors.initials: J
authors.lastname: Pierce
authors.forename: Ian
authors.initials: I
authors.lastname: Nicoud
authors.forename: Andrey
authors.initials: A
authors.lastname: Belous
authors.forename: Clayton D
authors.initials: CD
authors.lastname: Knox
authors.forename: Ravi S
authors.initials: RS
authors.lastname: Chari
mesh_terms: D000410:Alanine Transaminase / Q000032:analysis / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D024442:Aspartate Aminotransferase, Mitochondrial / Q000032:analysis / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D004195:Disease Models, Animal; D007511:Ischemia / Q000473:pathology / Q000628:therapy*; D008099:Liver / Q000098:blood supply* / Q000473:pathology; D008102:Liver Circulation / Q000502:physiology; D008297:Male; D009336:Necrosis / Q000473:pathology; D011336:Probability; D051381:Rats; D017207:Rats, Sprague-Dawley; D015424:Reperfusion / Q000379:methods; D015427:Reperfusion Injury / Q000517:prevention & control*; D012307:Risk Factors; D012430:Ruthenium Red / Q000494:pharmacology*; D012680:Sensitivity and Specificity
chemical_list: D012430:Ruthenium Red; D024442:Aspartate Aminotransferase, Mitochondrial; D000410:Alanine Transaminase; D002118:Calcium