Hepatitis C Virus NS5A and Core protein induce hepatic stellate cells activation promoting fibrosis-related gene regulation on hepatocytes.

Abstract

Introduction and Objectives: Chronic HCV infection leads to the development of liver fibrosis mediated by intercellular communication between hepatocytes and hepatic stellate cells (HSCs). The diverse molecular pathways involved in the development of fibrosis in hepatocytes derived from HSC activation induced by viral proteins remain to be fully determined. Our aim is to determine differentially expressed genes associated with fibrotic processes in hepatocytes (Huh7) that express the HCV NS5A or Core protein during co-culture with HSC (LX2). Materials and methods: Huh7 cells were transfected to express NS5A or Core proteins and co-cultured with HSC-LX2 cells. Viral protein expression and expression of TGFβ1, Col1, and aSMA was determined to assess LX2 activation. A profile of 84 genes associated with fibrosis during co-cultivation was determined and analyzed. Results: HSC-LX2 co-cultured with transfected Huh7 showed an 8.3, 6.7 and 4-fold increase in collagen1, TGFB1 and timp1 expression respectively induced by NS5A and a 6.5, 1.8 and 6.2-fold increase respectively induced by Core, all these compared to HSC-LX2 co-cultured with untransfected Huh7. We detected 28 overexpressed genes in Huh7 (NS5A+) and 46 differentially expressed genes in Huh7 (Core+) in co-culture with HSC-LX2, compared to untransfected Huh7 in co-culture with HSC-LX2. Analysis of the expression profile showed that the TGFβ1, the ECM regulation, and growth factors pathways are the molecular mechanisms involved during the co-culture of Huh7 transfected with NS5A or Core with HSC-LX2. Conclusions: HCV NS5A and Core proteins expression in Huh7 cells induces the HSC-LX2 activation, regulating the expression of diverse genes in hepatocytes that trigger different molecular mechanisms involved in the fibrosis development, this information provide the identification of possible anti-fibrotic targets drugs associated with HCV infection for further study.