Restricted redox oscillation in oxidative phosphorylation in jaundiced rat liver mitochondria and its relation to calcium ion.

High morbidity and mortality in surgical management for patients with obstructive jaundice was greatly attributed to the metabolic derangements in jaundiced liver mitochondria. Isolated liver mitochondria from jaundiced rat, produced by common bile duct ligation, were used to study the relationship among NADH level, oxygen consumption, and extramitochondrial calcium concentration. Alterations in NADH percentage and oxygen consumption were accomplished by incubating mitochondria with different substrates and monitoring oxygen consumption and NAD(P)H fluorescence simultaneously. In jaundiced liver mitochondria with glutamate + malate as substrate, respiration increased after the addition of exogenous Ca2+ at concentrations of 1 x 10(-7), 5 x 10(-7), and 1 x 10(-6) M. The maximal effect occurred at 5 x 10(-7) M. With different NADH-related substrates, the NAD(P)H fluorescence measurements (X axis) correlated linearly with state 3 respiration (Y axis), the slopes of the correlation curves being 2.27 and 0.79 in control and jaundiced mitochondria, respectively. After the addition of 5 x 10(-7) M Ca2+, the respirations of both control and jaundiced mitochondria increased and the slope for jaundiced mitochondria rose to 1.67. The matrix free Ca2+ concentration in jaundiced mitochondria, measured by fluo-3 loading, was higher than that in controls (162.1 +/- 16.7 nM, vs 129.7 +/- 12.6 nM, P < 0.01), while the matrix free/total Ca2+ ratio decreased from 34.9 +/- 6.0 (x10(-6)) to 27.2 +/- 4.4 (x10(-6), 9- < 0.05. The amplitude of the change in NAD(P)H fluorescence was reduced in jaundiced rat liver mitochondria and this correlated with the depression of respiration. A decrease in free/total Ca2+ ratio may be closely related to mitochondrial respiratory impairment in jaundice.