Mitochondrial dysfunction during anoxia/reoxygenation injury of liver sinusoidal endothelial cells.

Abstract

Sinusoidal endothelial cell injury plays a pivotal role in anoxia/reoxygenation liver damage. However, the mechanisms culminating in anoxia/reoxygenation endothelial cell injury remain unclear. Our aims were to determine whether anoxia/reoxygenation injury of sinusoidal endothelial cells causes mitochondrial dysfunction. In cultured rat liver sinusoidal endothelial cells, the mitochondrial membrane potential, cytosolic free calcium and cytosolic pH were quantitated by means of fluorescent probes and multiparameter digitized video microscopy. Cell viability was measured on the basis of lactate dehydrogenase release, and ATP was quantitated with a luciferin/luciferase assay. Mitochondrial membrane potential was stable during 90 min of aerobic perfusion. After 60 and 90 min of anoxia, mitochondrial membrane potential decreased gradually to 97% +/- 6% and 79% +/- 7% of the basal value, respectively. However, mitochondrial membrane potential decreased abruptly with reoxygenation after 60 min of anoxia to 45% +/- 12% of the basal value and did not recover over 30 min of aerobic perifusion. Loss of mitochondrial membrane potential could not be attributed to changes of cytosolic free calcium, cytosolic pH, nitric oxide generation or activity of poly(ADP-ribose) polymerase.(ABSTRACT TRUNCATED AT 250 WORDS)