NF-κB Signaling May Contribute To The Differentiation Of BMSCs Into Hepatocyte-like Cells

Objective: To investigate whether nuclear factor-kappa B(NF-кB) signaling pathway participated the process of differentiation of bone mesenchymal stem cells into hepatocyte-like cells induced by Hepatocyte growth factor(HGF), beta-nerve growth factor(β-NGF) and liver tissue fluid(LTF).Methods: BMSCs were isolated from bone marrow of adult rats and cultured in low-glucose DMEM with 10% fetal bovine serum(FBS) for expanding. Then BMSCs at third generation were divided into two main group: induced group and inhibition group which added BAY 11-7082(NF-кB inhibitor) more than induced group, Each main group was comprised of control group, hepatocyte growth factor(HGF) group, beta-nerve growth factor(β-NGF) group, liver tissue fluid(LTF) group, and respectively cultured in the presence of equal medium, HGF, β-NGF, liver tissue fluid. The feature of differentiated hepatocytes was identified by indocyanine green(ICG) for the hepatocytes transferring function and by mmunofluorescence for alpha-1-antitrypsin(AAT) which is the marker of hepatocytes. Western blot technique was used to detect the protein expressions of NF-кB(p65)and AAT.Results: The majority of differentiated cells from HGF induced group, β-NGF induced group and LTF induced group were showed spherical or ovate in shape, ICG and AAT were positive, the NF-кB transferred into nuclear. Although inhibition HGF group,inhibition β-NGF group and inhibition LTF group were showed fusiform or polygonal morphologies unchanged in shape, ICG uptake and NF-кB positive rates were reduced,the AAT expression was significantly decreased.Concludions: HGF, β-NGF and LTF could induce the differentiation of BMSCs into hepatocyte-like cells which expressed hepatocyte marker, AAT protein in cytoplasm and uptook ICG and had the morphologic features. During the inducing process, NF-кB signaling may contribute to the differentiation of BMSCs into hepatocyte-like cells.