Abstract
Background: Proton leak (H+ leak) dissipates mitochondrial membrane potential (mΔΨ) through the re-entry of protons into the mitochondrial matrix independent of ATP synthase. Changes in H+ leak may affect reactive oxygen species (ROS) production. We measured H+ leak and ROS production during ischemia-reperfusion and ischemic preconditioning (IPC) and examined how changing mitochondrial respiration affected mΔΨ and ROS production. Materials and Methods: Isolated rat hearts (n = 6/group) were subjected to either control-IR or IPC. Rate pressure product (RPP) was measured. Mitochondria were isolated at end reperfusion. Respiration was measured by polarography and titrated with increasing concentrations of malonate (0.5-2 mM). mΔΨ was measured using a tetraphenylphosphonium electrode. H+ leak is the respiratory rate required to maintain membrane potential at -150 mV in the presence of oligomycin-A. Mitochondrial complex III ROS production was measured by fluorometry using Amplex-red. Results: IPC improved recovery of RPP at end reperfusion (63% ± 4% versus 21% ± 2% in control-IR, P <0.05). Ischemia-reperfusion caused increased H+ leak (94 ± 12 versus 31 ± 1 nmol O/mg protein/min in non-ischemic control, P <0.05). IPC attenuates these increases (55 ± 9 nmol O/mg protein/min, P <0.05 versus control-IR). IPC reduced mitochondrial ROS production compared with control-IR (31 ± 2 versus 40 ± 3 nmol/mg protein/min, P <0.05). As mitochondrial respiration decreased, mΔΨ and mitochondrial ROS production also decreased. ROS production remained lower in IPC than in control-IR for all mΔΨ and respiration rates. Conclusions: Increasing H+ leak is not associated with decreased ROS production. IPC decreases both the magnitude of H+ leak and ROS production after ischemia-reperfusion.
Original language | English (US) |
---|---|
Pages (from-to) | 5-14 |
Number of pages | 10 |
Journal | Journal of Surgical Research |
Volume | 165 |
Issue number | 1 |
DOIs | |
State | Published - Jan 2011 |
Externally published | Yes |
Keywords
- Heart
- Ischemia
- Mitochondria
- Proton leak
- Reactive oxygen species
- Reperfusion
- Uncoupling
ASJC Scopus subject areas
- Surgery