- 作者：Marí ; a Victoria Garcí ; a-Mediavilla ; Sonia Sá ; nchez-Campos ; Pilar Gonzá ; lez-Pé ; rez ; Marta Gó ; mez-Gonzalo ; Pedro Lorenzo Majano ; Manuel Ló ; pez-Cabrera ; Gerardo Clemente ; Carm
- 刊名：Journal of Hepatology
- 出版年：2005
- 出版时间：2005
- 年：2005
- 卷：43
- 期：4
- 页码：606-613
- 全文大小：284 K
lass=""h4"">Background/Aims
We aimed to explore the effects of hepatitis C virus (HCV) core and NS5A proteins on reactive oxygen (ROS) and nitrogen species (RNS) formation and on gene expression profile of iNOS in human hepatocyte-derived cells.lass=""h4"">Methods
Production of ROS and RNS and nitrotyrosine residues accumulation were determined by flow cytometry and fluorescent microscopy as well as by Western blot, respectively, in NS5A- and core-transfected cells. Northern blot, Western blot, real-time PCR, and luciferase assays were used to assess iNOS gene expression in both transfectants.
lass=""h4"">Results
Cytokine-activated NS5A- and core-transfected cells induced ROS and RNS production but an earlier and more marked increase was observed in NS5A-expressing cells. Superoxide production was also augmented, showing a similar temporal pattern of appearance in both NS5A- and core-transfected cells. Although both NS5A and core HCV proteins were able to up-regulate iNOS gene expression, accompanied by a nitrotyrosine-containing proteins accumulation, an earlier iNOS overexpression was observed in NS5A-expressing cells, suggesting a different time course of iNOS activation pattern for core and NS5A HCV proteins.
lass=""h4"">Conclusions
Our results indicate a differential contribution of both HCV proteins to oxidative and nitrosative stress generation.