Effect of short time ischemia/reperfusion in vivo on rat kidney mitochondria oxidative phosphorylation

Author: Lukas Mickevičius Title of the paper: Effect of short time ischemia/reperfusion in vivo on rat kidney mitochondria oxidative phosphorylation The aim of this research: To investigate an effect of short time ischemia/reperfusion in vivo on rat kidney mitochondria oxidative phosphorylation. Objectives: To evaluate the effect of 20 min ischemia and 30 min reperfusion on mitochondria oxidative phosforilation system and investigate rat mitochondrial respiration chain complex I, II and II + III activity. Object of this research: Wistar breed rats males were used to perform this research. Methods: Warm ischemia (37 ° C) to rat kidneys was induced by clamping renal arteries using vascular clamps. Ischemia was induced for 20 min and after that reperfusion lasted for 30 min. Kidneys were removed and mitochondria were isolated by using differential centrifugation method. The amount of proteins was measured via Buret method. Mitochondrial respiration rates were measured by Oxygraph-2k system and using glutamate/malate and succinate as substrates. Mitochondrial respiration chain complexes activity was measured spectrophotometrically. Results: This research results show that short time (20 min) ischemia and reperfusion (30 min) does not affect the respiration rates when mitochondrial respiration chain complex I substrate glutamate/malate is being oxidized. This research shows that oxidizing mitochondrial respiration chain complex II substrate succinate evaluates respiration rate in state two after short-time ischemia 1.47 times but didn’t affect state three. Oxidizing succinate respiration control index decreases by 22 % which show that even after short-time ischemia mitochondrial membrane is getting damaged. Complex I activity decreased by 67% after 20 min ischemia and 30 min reperfusion.. Conclusions: Research showed that even short time of ischemia damages mitochondrial oxidative phosphorylation system. Short-time ischemia decreases mitochondrial respiration chain complex