Cold preservation-warm reoxygenation increases hepatocyte steady-state Ca(2+) and response to Ca(2+)-mobilizing...

Although the role of Ca(2+) in liver transplantation injury has been the object of several studies, direct evidence for alterations in intracellular Ca(2+) homeostasis after cold preservation-warm reoxygenation (CP/WR) has never been presented. We thus investigated the effects of CP/WR on steady-state Ca(2+) and responses to a Ca(2+)-mobilizing agonist. Isolated rat hepatocytes were suspended in University of Wisconsin solution, stored at 4 degrees C for 0, 24, and 48 h, and reoxygenated at 37 degrees C for 1 h. Cytosolic Ca(2+) was measured in single cells by digitized fluorescence videomicroscopy. CP/WR caused a significant increase in steady-state cytosolic Ca(2+), which was inversely proportional to cell viability. Pretreatment of hepatocytes with an agent that protects mitochondrial function attenuated the increase in steady-state cytosolic Ca(2+) and improved hepatocyte viability. Ca(2+) responses to the purinergic agonist ATP also increased significantly as a function of cold storage time. This increase was related to an increase in the size of inositol 1,4,5-trisphosphate-sensitive Ca(2+) stores and subsequent capacitative Ca(2+) entry. Thus CP/WR significantly perturbs steady-state hepatocellular Ca(2+) and responses to Ca(2+)-mobilizing agonists, which may contribute to hepatocyte metabolic dysfunction observed after CP/WR.