An Nrf2-NF-kB cross-talk controls hepatocyte proliferation in the normal and injured liver

The liver has remarkable regenerative and detoxification capacities, which require the Nrf2 and NF-kB transcription factors. Although their individual functions in hepatocytes are well characterized, knowledge about their cross-talk in the adult liver is limited. Therefore, we deleted the genes for Nrf2, the NF-kB subunit p65 or both genes in adult hepatocytes of mice using AAV8-Cre viruses. Loss of p65 or both genes caused spontaneous liver inflammation and necrosis, combined with a progressive increase in hepatocyte proliferation. Gene expression profiling identified individual and common target genes of both transcription factors, including genes involved in the control of cell proliferation. Overall design: Control (C57BL6/j), Nrf2 flox/flox, p65 flox/flox and Nrf2:p65 double floxed mice were injected with AAV8Cre to induce a hepatocyte specific knockout of the respective genes. 12 days after injection, mice were euthanized and the liver was perfused with Liberase TM solution followed by low speed centrifugation of the cell suspension and additional centrifugation in Percoll density media. Total RNA was isolated from freshly-isolated hepatocytes and subjected to library preparation using the TrueSeq stranded protocol. After single end sequencing in an Illumina Novaseq machine, reads were aligned to the mouse genome GRCm39 using Salmon softwere. Differential expression was done using DESeq2.