Hyperoxia and thyroxine treatment and the relationships between reactive oxygen species generation, mitochondrial membrane potential, and cardiolipin in human lens epithelial cell cultures

Thyroxine-treated human lens epithelial cells, HLE B-3, were grown in either a normoxic or hyperoxic atmosphere as a first step in identifying factors related to their increased viability. Reactive oxygen species, ROS, and the apparent mitochondrial membrane potential, Delta Psi(m), were measured using fluorescent probes. ROS were significantly higher in HLE B-3 cells grown in a hyperoxic atmosphere for both thyroxine-treated and untreated samples. However, treatment with thyroxine produced 40% lower ROS levels than untreated cells. A mitochondrial uncoupler concomitantly reduced ROS generation. In cells that were grown in a hyperoxic atmosphere, the Delta Psi(m) was significantly higher for samples treated with thyroxine compared to those that were untreated. ROS generation correlated inversely with the apparent Delta Psi(m) and the amount of cardiolipin, and correlated with the amount of lipid oxidation products. These correlations were valid whether cardiolipin and the Delta Psi(m) were decreased as a result of oxygen or increased as a result of thyroxine treatment. Therapies that protect mitochondria from damage and reduce damaging ROS generation may potentially ameliorate the effects of oxidation that occur in aging tissues and in diseases such as cataract.