New Technique for Studying Liver Fibrogenesis

Researchers in France have devised a way to reproduce liver fibrogenesis related to HCV infection, or exposure to ethanol of lipids.

In an abstract to be presented at the International Liver Congress in Barcelona, S. Lagaye of the Institut Pasteur and colleagues took non-infected liver slices from human liver slices and cultivated them.

They infected the tissue with HCVcc supernatant both with and without EtOH or palmitate 500.

They evaluated markers of fibrosis in uninfected liver slices as control and in HCV-infected slices.

Those were :expression of Tumor growth factor b (TGFβ), Heat shock protein 47 (Hsp47), Alpha smooth muscle actin (αSma), Procollagen1 A1 (Procl1A1), Matrix metalloproteinases 2, 9 (MMP-2, 9), Vascular Endothelial Growth Factor (VEGF), which were checked by RTqPCR. Levels of TGFb, Triglycerides, Aldehyde deshydrogenase (ALDH), Alcohol Deshydrogenase (ADH) activity and viability assays [ATP (%)], were determined by ELISA assays.

At day 21, NI LS and INF LS had a viability rate of 75% and 68%, respectively. In the presence of EtOH, TGFβ, Hsp47, αSma, Procl1A1, MMP-2, MMP-9, VEGF expression and triglyceride production in NI and INF LS increased in a dose-dependent manner and to a higher extent in INF LS as compared to NI LS [at day 21, in INF LS treated with 25 mM EtOH: a significant 3, 3.5, 3.6, 2.6, 3.5, 4, 4.6, 2 fold increase as compared to controls, respectively].

At day 21, in INF LS treated, MMP- 9 expression was higher than MMP-2 expression [×2]. Addition of palmitate increases significantly TGFb, Hsp47, αSma, Procl1A1, MMP- 2, 9, VEGF expression and to a higher extent in INF LS than in NI LS [at day 21, in INF LS treated: a significant 5, 7, 4, 3.8, 4.8, 4, 4.7 fold increase, respectively as compared to CTRL] with a triglyceride production higher in INF LS [∼×2.75].

At day 21, MMP-2 expression in NI and INF LS was higher with palmitate treatment [×2.7, 2.5 respectively] than with EtOH treatment. HCV infection with EtOH or palmitate seems to have an additional effect on the expression of the fibrosis markers.

The team concluded that in this ex vivo model, supporting hepatocyte-specific gene expression for 21 days, reproduces the liver fibrogenesis related to HCV infection, ethanol or lipids exposure.

Their method “could be an easy tool for studying human liver fibrogenesis and evaluating the potency of new antifibrotic therapies in the human liver tissue,” they concluded.