Previous studies have demonstrated that NF-κB (nuclear factor κB) pathway is involved in promoting cell proliferation. To further explore the regulatory branches and their sequence of NF-κB pathway in promoting hepatocyte proliferation at transcriptional level during rat liver regeneration (LR), Rat Genome 230 2.0 Array was used to detect the expression changes of the isolated hepatocytes. We found that many genes involved in NF-κB pathway (including 73 known genes and 19 homologous genes) and in cell proliferation (including 484 genes and 104 homologous genes) were associated with LR. Expression profile function (Ep) was used to analyze the biological processes. It showed that NF-κB pathway promoted hepatocyte proliferation through three branches. Several methods of integrated statistics were applied to extract and screen key genes in LR, and it indicated that eight genes may play a vital role in rat LR. To confirm the above predicted results, the content of proteins ,such as CCND1 and DUSP1 were analyzed using two-dimension gel electrophoresis and MALDI-TOF/TOF, and the results were generally consistent with that of microarray data. It is concluded that 3 branches and 8 key genes involved in NF-κB pathway regulate hepatocytes proliferation during rat liver regeneration.