Depolarized mitochondrial membrane potential and protection with duroquinone in isolated perfused lungs from rats exposed to hyperoxia

Dissipation of mitochondria! membrane potential (Delta psi(m)) is a hallmark of mitochondrial dysfunction. Our objective was to use a previously developed experimental-computational approach to estimate tissue Delta psi(m) in intact lungs of rats exposed to hyperoxia and to evaluate the ability of duroquinone (DQ) to reverse any hyperoxia-induced depolarization of lung Rats were exposed to hyperoxia (>95% O-2) or normoxia (room air) for 48 h, after which lungs were excised and connected to a ventilation-perfusion system. The experimental protocol consisted of measuring the concentration of the fluorescent dye rhodamine 6 G (R6G) during three single-pass phases: loading, washing, and uncoupling, in which the lungs were perfused with and without R6G and with the mitochondrial uncoupler FCCP, respectively. For normoxic lungs, the protocol was repeated with 1) rotenone (complex I inhibitor), 2) rotenone and either DQ or its vehicle (DMSO), and 3) rotenone, glutathione (GSH), and either DQ or DMSO added to the perfusate. Hyperoxic lungs were studied with and without DO and GSH added to the perfusate. Computational modeling was used to estimate lung Delta psi(m) from R6G data. Rat exposure to hyperoxia resulted in partial depolarization (-33 mV) of lung Delta psi(m) and complex I inhibition depolarized lung Delta psi(m )by -83 mV. Results also demonstrate the efficacy of DQ to fully reverse both rotenone- and hyperoxia-induced lung Delta psi(m) depolarization. This study demonstrates hyperoxia-induced Delta psi(m) depolarization in intact lungs and the utility of this approach for assessing the impact of potential therapies such as exogenous quinones that target mitochondria in intact lungs. NEW & NOTEWORTHY This study is the first to measure hyperoxia-induced Delta psi(m )depolarization in isolated perfused lungs. Hyperoxia resulted in a partial depolarization of Delta psi(m), which was fully reversed with duroquinone, demonstrating the utility of this approach for assessing the impact of potential therapies that target mitochondria such as exogenous quinones.

Automated summary

This study used an experimental-computational approach to estimate tissue Delta psi(m) in intact lungs of rats exposed to hyperoxia, and evaluated the ability of duroquinone (DQ) to reverse any hyperoxia-induced depolarization. It was found that exposure to hyperoxia resulted in depolarization of lung Delta psi(m), and DQ effectively reversed this depolarization. This study is the first to measure hyperoxia-induced Delta psi(m) depolarization in isolated perfused lungs.