NF-κB participates in hepcidin up-regulation induced by iron overload in HH4 hepatocytes
10.3969/j.issn.1000-4718.2015.04.022
- VernacularTitle:NF-κB 参与枸橼酸铁铵过载诱导的人肝细胞 hepcidin 基因表达
- Author:
Shiwei LI
;
Xiang LI
;
Feng GUAN
- Publication Type:Journal Article
- Keywords:
Nuclear factorκB;
Chromatin immunoprecipitation;
Electrophoretic mobility shift assay;
Dual-lu-ciferase reporter assay system;
Hepcidin
- From:
Chinese Journal of Pathophysiology
2015;33(4):695-701
- CountryChina
- Language:Chinese
-
Abstract:
[ ABSTRACT] AIM:To study the effects of nuclear factor kappa B ( NF-κB) on human hepcidin expression in fer-ric ammonium citrate ( FAC)-induced HH4 hepatocytes.METHODS:Non-transformed HH4 cells were exposed to FAC at concentrations of 0.1, 1, 5 and 10 mmol/L for 48 h.The expression of iron regulatory gene hepcidin was determined by semi-quantitative RT-PCR.The effects of NF-κB on hepcidin transcriptional activity were detected using chromatin immuno-precipitation (ChIP), electrophoretic mobility shift assay (EMSA) and dual-luciferase reporter assay system, combined with the inhibition experiments of intracellular NF-κB activity.RESULTS: FAC at concentrations of 5 mmol/L and 10 mmol/L significantly enhanced the expression of hepcidin.The results of ChIP and EMSA showed the binding of NF-κB to the upstream of hepcidin promoter.Treatment with NF-κB inhibitor BAY 11-7082 attenuated hepcidin expression.The lucif-erase activity in the cells transfected with recombinant luciferase reporter plasmid was obviously higher than that in control group.CONCLUSION:NF-κB is the transcription factor that contributes to hepcidin expression in iron overload-induced HH4 cells.
