NO Signal Up-regulates The Expression Of Endogenous F? In Human Hepatocytes

OBJECTIVE:To investigate the effects of histone acetylation and deacetylation on the promoter region of coagulation factor ?(F?)in normal human hepatocytes of human immortalized normal liver cells.To explore the molecular mechanism of NO signaling in the activation of F? in LO2 cells.Method:Adding L-arginine to the culture medium of human liver cells L02 in vitro,to make L02 express F?.The L02 cells at logarithm growth phase were selected and randomly divided into blank control group and L-arginine group,the C646 group and the SAHA group respectively.L-arginine group(final concentration of L-arginine 10mM),The control group replaced L-arginine with the same volume of PBS,the C646 group(final concentration of C646 2uM),the SAHA group(final concentration of SAHA 50nM).culture 0h,12 h,24h,36 h,48h,60 h.Use RT-PCR method to detect the transcript levels of human F? gene in each group.Detection of histone acetylation level in F? promoter region by chromatin immunoprecipitation(ChIP).And detection NF-kB1 and F? gene promoter region binding,F? gene promoter region histone acetylation level.Results:The results of RT-PCR showed that the expression of F? was up-regulated in L-arginine group and the SAHA group,The F? gene was low expressed in the control group and the C646 group.ChIP detection NF-kB1 and F? gene promoter region binding,F? gene promoter region histone acetylation level increased.Control group and the C646 group F? gene promoter region histone acetylation level for the background expression,L-arginine group and the SAHA group F? gene promoter region histone acetylation level improved.Conclusion:The histone acetylation inhibitor can abrogate the up-expression of human F? gene.Histone deacetylation inhibitors enhance the up-expression of human F? gene.NO signaling in LO2 cells by regulating the F? gene promoter region histone acetylation,recruiting transcription factor NF-kB1 activated the expression of the human F? gene.