The Study Of Romidepsin In Reducing Liver Fibrosis Via NF-?b Signaling Pathway

Objective:To observe the effect of Histone Deacetylase inhibitor Romidepsin?FK228?on liver cirrhosis in mice,and to investigate the underlying mechanism of renoprotective effect of FK228.Methods:1.In vivo:Thirty experimental mice were randomly divided into three groups,control group?group A?,model group?group B?and treatment group?group C?.To set up the liver cirrhosis model,mice in group B and C were treated with intraperitoneal injection of CCl4 for 24 weeks.From the21^th week,group C were treated with intraperitoneal injection of FK228weekly.The levels of serum ALT and AST were detected to judging liver damage degree;HE and Sirius Red staining were used to observe pseudolob?li and tubercle.Immunohistochemical staining and Western blot analysis were used to detect the expression of osteopontin?-SMA in liver.2.In vitro:human hepatic stellate cells LX-2 were c?ltivated,and randomly divided into four groups:normal control group?PBS?,stim?lation group?5ng/ml TGF-?1?,low dose treatment group?5ng/ml TGF-?1+0.5?mol/ml FK228?and high-dose treatment group?5 ng/ml TGF-?1+1?mol/ml FK228?.RT-PCR was used to detect the expression of stellate cell-associated activator proteins gfap,desmin,col?1 and col1?4;Western blot was used to test the levels of total protein?-SMA,P65,and pP65;immunofluorescence was used to observe transfer into nuclear of P65.Results:?1?In vivo:The data showed the average serum ALT in group C was37.67±8.4611 IU/L and 67.27±11.109 IU/L in group B.It is significant decreased when treat the experimental mice with FK228?P=0.019?.Also,H&E and Sirius Red staining revealed the degree of liver fibrosis in group C were weakened obviously.Additionally,Western Blotting and Immunohistochemistry indicate the levels of?-SMA in group C were notable reduced?P=0.001?.?2?In vitro:When stimulated by TGF-?1,mRNA of stellate cell activation-relatedgenesdesmin?2.87?0.32?,gfap?5.57?0.87?,col1?1?5.27?1.01?and col3?1?6.31?0.32?were significantly increased?P<0.05?.FK228 obviously inhibited the activation of hepatic stellate cells induced by TGF-?1?P<0.05?.Western blot detection of?-SMA was also verified.At the same time,the expression level of pP5 was significantly increased.The cellular immunofluorescence revealed that P65in the stimulation group was largely nucleated,and FK228 could inhibit its migration to the nucleus.Conclusion:The results of this study firstly prove the preventive effect of FK228treatment for liver fibrosis and the mechanism may involves inhibition of NF-?B signaling pathway.