Artesunate Inhibits HCV Replication In Cell Models

Background and Objective:Hepatitis C virus (HCV) is globally distributed. HCV infection increased very fast, and though the treatment for HCV has obtained great achievements, but due to the current treatment methods have limitations, new methods and drugs for hepatitis C treatment is still being developed. In this study, JFH1infected Huh7.5.1and OR6cell models were used for artesunate, ribavirin and interferon to affect HCV replication.Methods:By MTT to detect the Huh7.5.1and OR6cell proliferation. HCVRNA fluorescence quantitative PCR test was used to detect HCVRNA. Luciferase assay was applied to evaluate the luciferase activity. Western blot testing was employed to the effect of artesunate on HCV NS5A protein expression.Result:1、Artesunate suppresses HCVRNA replication in JFH1infected Huh7.5.1cells. Artesunte decreases HCVRNA1～21og10IU/ml in the first week and2-31ogIU/ml in the second week. RBV10ug/ml only decreases HCVRNA1logIU/ml until the second week. IFN-a2b500U/ml decreases HCVRNA41og10IU/ml in the first week and was undetectable in the second week.2、Artesunate declines HCV coupled renilla luciferase activity in OR6cells.fluorescence activity of the treatment groups are listed as followings:Blank:1±0.00; A12.5umol/L:0.159±0.02; A25umol/L:0.084±0.00; While RBV10ug/ml:0.776±0.01; IFN-a2b500U/ml:0.014±0.00; A12.5umol/l+IFN-a2b500U/ml:0.001±0.00。Compared to blank group, the dataset showed significant (p<0.05)。3、Artesunte reduces NS5A protein expression in OR6cells. The relative intensity rations of NS5A/β-actin in each group follows as below:blank:1.084±0.074; A12.5umol/L:0.377±0.01; A25umol/L:0.316±0.01; RBV10ug/ml:0.922±0.10; IFN-a2b500u/ml:0.180±0.00; A12.5umol/L+IFN-a2b500U/ml: 0.124±0.01。Compared to blank group, the dataset showed significant (p<0.05).4、Artesunte upregulation the activity of HCV5’UTR, The relative luciferase activity in each group follows as below:blank:7.510±0.098; A12.5umol/L,28.979±1.595; A25umol/L,49.249±1.502; A37.5umol/L,60.877±1.572而RBV10ug/ml:7.335±0.095; IFN-a2b500U/ml:2.035±0.025。Compared to the blank group, except the rebaviran group,the other dataset showed significant。Conclusion:Artesunate inhibits HCVRNA replication and HCVNS5A protein expression in a dose and time dependent way. The inhibitive effect on HCV replication is stronger than ribavirin, but weaker than interferon. Artesunate has the synergistic inhibitive effect on HCV with interferon。Artesunate inhibiting HCV replication is not by influencing the activity of5’UTR, there may be other mechanisms.