Indirubin-3'-oxime prevents hepatic I/R damage by inhibiting GSK-3beta and mitochondrial permeability transition.
Indirubin-3'-oxime is an indirubin analogue that shows favorable inhibitory activity targeting glycogen synthase kinase 3beta (GSK-3beta). In this study, we evaluated if acute treatment with indirubin-3'-oxime (Ind) prevents hepatic ischemia/reperfusion (I/R) damage. Wistar rats were subjected to 150 min of 70% warm ischemia and 16 h of reperfusion. In the treated group 1 microM indirubin-3'-oxime was administered in the hepatic artery 30 min before ischemia. Acute treatment with Ind decreased serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) levels, comparatively to I/R livers. Bax translocation to the mitochondria and cytochrome c release were higher in I/R livers. Ind treatment significantly attenuated Bax translocation and preserved mitochondrial cytochrome c content. Ind also protected mitochondria from calcium-induced mitochondrial permeability transition (MPT), as well as the decrease in state 3 mitochondrial respiration, the delay in the repolarization after a phosphorylative cycle and the decrease in ATP content caused by I/R. By addressing GSK-3beta activity and phosphorylated GSK-3beta at Ser(9) content in liver homogenates and isolated mitochondria, data suggests that inhibition of GSK-3beta by indirubin-3'-oxime prevents the increase in mitochondrial phosphorylated GSK-3beta at Ser(9) induced by I/R, thus correlating with MPT inhibition and preservation of cytochrome c content. Pre-treatment with indirubin-3'-oxime in conditions of hepatic I/R, protects the liver by maintaining mitochondrial function and hepatic energetic balance.
Varela
Ana T
AT
Center for Neurosciences and Cell Biology, Department of Zoology, University of Coimbra, 3004-517 Coimbra, Portugal.
Simões
Anabela M
AM
Teodoro
João S
JS
Duarte
Filipe V
FV
Gomes
Ana P
AP
Palmeira
Carlos M
CM
Rolo
Anabela P
AP
D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000008:administration & dosage*; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors*; D000071679:Glycogen Synthase Kinase 3 beta; D007211:Indoles / Q000008:administration & dosage*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000187:drug effects*; D008111:Liver Function Tests; D008297:Male; D008928:Mitochondria / Q000187:drug effects*; D010091:Oximes / Q000008:administration & dosage*; D010539:Permeability / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000517:prevention & control*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
C494828:Bax protein, rat; D004791:Enzyme Inhibitors; D007211:Indoles; D010091:Oximes; D051028:bcl-2-Associated X Protein; C474021:indirubin-3'-monoxime; D045304:Cytochromes c; D007770:L-Lactate Dehydrogenase; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D000071679:Glycogen Synthase Kinase 3 beta; C000605508:Gsk3b protein, rat; D038362:Glycogen Synthase Kinase 3