Oxygen dependence and subcellular partitioning of hepatic menadione-mediated oxygen uptake. Studies with isolated hepatocytes, mitochondria, and microsomes from rat liver in an oxystat system

Using an oxystat system, menadione (2-methyl-1,4-naphthoquinone)-mediated oxygen uptake was investigated in isolated rat hepatocytes, in malate/glutamate-supplemented mitochondria, and in NADPH-reduced microsomes at steady-state oxygen partial pressures (pO2) between 0.1 to 100 mm Hg (0.2-150 microM O2). Menadione-mediated stimulation of oxygen uptake was half-maximal at pO2 of 0.5, 0.2, and 0.9 mm Hg, respectively. In hepatocytes and mitochondria half-maximal concentrations of menadione were 15 and 4 microM. However, in microsomes saturation with menadione was not reached at concentrations up to 300 microM. Antimycin A inhibited menadione-mediated oxygen uptake in hepatocytes and mitochondria by about three-fourths, while rotenone was without inhibitory effect; KCN inhibited practically completely. In mitochondria menadione-stimulated oxygen uptake was significantly inhibited by dicoumarol but further enhanced by the addition of ADP, even in the presence of rotenone. The results suggest that menadione-mediated hepatocellular oxygen uptake proceeds almost independently of pO2 in most regions of the liver lobule but that in areas of low pO2 such as the centrolobular regions limitation by oxygen may occur. They also demonstrate that in the intact hepatocyte menadione-mediated oxygen uptake predominantly (greater than 90%) results from electron transfer in the mitochondrial respiratory chain by menadione.