关键词: 关键词：枸橼酸铁铵, 丙型肝炎病毒, 内部核糖体进入位点, 活性氧

Abstract: Objective The major goal of this study was to identify the activity of HCV IRES translation differences and identify the relationship between HCV IRES translation activity and ROS Under different concentrations of FAC induction. Methods 1) Expression plasmid pCI-Rluc-HCV IRES-Fluc was confirmed by endonuclease digestion as well as luciferase transient expression in Huh-7 cell;2) Controlled by Dual-Luciferase Reporter Assay,We analysis the Different translation activity of HCV internal ribosomal entry site (IRES) Under the concentration of 50 μmol/L and 300 μmol/L of FAC induction; ROS fluorescent staining method was used to detect the activity of ROS in cells Huh-7,Western blot method was used to Detect the protein expression changes of Nrf2 in cells Huh-7; 3) On the basis of the above experiments,we Add 100 μmol/L DPI in 300 μmol/L FAC experimental group,Analysis the changes of HCV replication and ROS production after joining DPI. Results The results showed that the generation of ROS and the activity of luciferase in the model group was significantly higher than that in the control group(P<0.05). FAC can enhance the expression of HCV IRES and Increase the production of ROS, then causing Nrf2 expression in Huh-7 cell. However,after adding ROS inhibitor DPI,The above functions in Huh-7 cell are weakened. Conclusions The increase of HCV IRES expression induced by FAC, related to excessive ROS production induced by FAC in Huh-7 cells.

Key words: Keywords: Ferric ammonium citrate, Hepatitis C virus, Internal ribosome-entry site, Reactive Oxygen Species