cytochrome-c-t and indirubin-3--monoxime

Zucker fatty rats were subjected to warm ischaemia and 12 hours of reperfusion. Ind or NAD. Pretreatment decreased markers of liver injury while preserving mitochondrial cytochrome c content, which is related to the prevention of calcium-induced mitochondrial permeability transition (mPT), the decline in mitochondrial respiratory state 3 and ATP content. The generation of reactive oxygen species (ROS) was also diminished. Inhibition of GSK-3ß by Ind resulted in the prevention of cyclophilin-D (CypD) phosphorylation, unabling it to bind to the adenine nucleotide translocator (ANT), thus, preventing mPT induction. Furthermore, deacetylation of CypD at Lys residue by sirtuin 3 (SIRT3) caused its dissociation from ANT, contributing to an increase in mPT threshold in NAD

Topics: Adenosine Triphosphate; Animals; Antibiotics, Antineoplastic; Calcium; Cyclophilins; Cytochromes c; Fatty Liver; Glycogen Synthase Kinase 3 beta; Hepatic Artery; Indoles; Liver; Mitochondria, Liver; Mitochondrial ADP, ATP Translocases; NAD; Oximes; Peptidyl-Prolyl Isomerase F; Rats; Rats, Zucker; Reactive Oxygen Species; Reperfusion Injury; Sirtuins; Warm Ischemia

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.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; bcl-2-Associated X Protein; Cytochromes c; Enzyme Inhibitors; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Indoles; L-Lactate Dehydrogenase; Liver; Liver Function Tests; Male; Mitochondria; Oximes; Permeability; Rats; Rats, Wistar; Reperfusion Injury