Volume 64 - 2001 - Fasc.3 - Original articles
Trichostatin A, lead compound for development of antifibrogenic drugs
Eukaryotic gene expression has mainly been studied in the context of trans-acting transcription factors and their interaction with regulatory cis-elements. Evidence is accumulating, that the higher order structure of chromatin also plays an essential role in eukaryotic gene expression. Hepatic stellate cells are the major cellular source of extracellular matrix synthesis in chronic liver diseases leading to fibrosis. We explored the antifibrogenic effect of the bistone deacetylase inhibitor trichostatin A (TSA) on hepatic stellate cells in vitro. Primary hepatic stellate cells as well as activated, subcultured stellate cells were exposed to 10-1 M - 10-1 M TSA. Collagens type I and 111, and smooth muscle a-actin (ct-SMA), a marker for transdifferentiation, were investigated at the protein and MRNA level by performing Northern hybridisation and quantitative immunoprecipitation. The antiproliferative effect was examined by IH-thymidine incorporation and cell counting. Hyperacetylation of historic H4 was demonstrated by acid urea Triton-X-100 (AUT) polyacrylamide gel electrophoresis. TSA at 101 M retarded the morphological changes characteristic for activation of primary stellate cells, Synthesis of coliagens type I and 111, and cc-SMA was strongly inhibited at both protein and mRNA level. The proliferation rate of primary hepatic stellate cells was strongly suppressed by 10-7 M TSA. Hyperacetylation of histone U4 showed to be maximal at IO-' M TSA. Primary hepatic stellate cells were more affected by TSA than subcultured stellate cells.
