Cyanidin 3-O-glucoside in protection against liver ischemia injury
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2013-10-08 |
Cyanidin 3-O-glucoside (Cy3G) is one of anthocyanins commonly found in berries and various vegetables. Our previous experiments showed that Cy3G is a potent agent to prevent ischemiainduced caspase activation in heart. Therefore the aim of this study was to investigate the effects of Cy3G on mitochondrial functions and caspase-3 activation in ischemic liver. The experiments were performed on 2-4 months old Wistar male rats (in vivo). Cy3G was injected through vena cava, and there was 15 min perfusion with the anthocyanin. After perfusion the liver were placed in the incubator at 37º C and subjected to 60 min total ischemia (autolysis). Cytosolic and mitochondrial fractions were separated by differential centrifugation. Mitochondrial respiration was registered using Oxygraph-2k. Hydrolysis of caspase-3 substrate was followed fluorimetrically. We found that Cy3G at tested concentrations (20 and 40 μM) had no significant effect on the routine (in the presence of glutamate and malate as substrates) and phosphorylating (State 3; in the presence of ADP) respiration rates of non-ischemic mitochondria. Interestingly, perfusion of liver with 40 μM but not with 20 μM of Cy3G prevented ischemia-induced mitochondrial dysfunction. We also found that perfusion of liver with Cy3G but only at a concentration of 20 μM prevented ischemia-induced caspase-3 activation. Thus, we show that Cy3G in a dose-dependent manner prevents ischemia-induced mitochondrial dysfunction and caspase-3 activation in liver. These results indicate the potency of Cy3G to regulate cell death in pathologies.