Cytochrome b5 Interacts With Cytochrome C and Inhibits Hepatocyte Apoptosis in Brain-dead Rabbit Donors

Abstract

BackgroundDonation after brain death (BD) liver grafts undergo the process of hypoxia-ischemia, which induces hepatocyte apoptosis, but the underlying mechanisms remain unclear. Cytochrome (Cyt) b5 expression was shown to be low in BD rabbits. This study aimed to investigate if Cyt b5 and Cyt c are involved in liver apoptosis after BD. Methods and resultsLiver tissue samples were obtained from donors after BD and from BD rabbit models. Tissues were analyzed by immunofluorescence, western blotting, and reverse-transcriptase polymerase chain reaction to detect Cyt b5 and Cyt c protein expressions and mRNA. Normal liver cells (LO-2) were cultured under serum deprivation and hypoxia, and analyzed as above. Cyt b5 protein and mRNA levels had decreased, while Cyt c levels had increased in BD liver donors and rabbits. Similar results were obtained in LO-2 cells cultured under hypoxia. After 6 and 12 hours of serum deprivation and hypoxia, apoptosis was increased, the levels of Cyt b5 gradually decreased, and the levels of Cyt c gradually increased over time; meanwhile, the Cyt b5-Cyt c combination was gradually reduced. A negative linear correlation between Cyt b5 and Cyt c was also observed. ConclusionsCyt b5 might be an anti-apoptotic protein that could protect the liver after BD and this protective effect might involve increased binding to Cyt c. This study provides some clues for improving the quality of donor livers.