title: S-Nitroso-N-acetylcysteine induces de-differentiation of activated hepatic stellate cells and promotes antifibrotic effects in vitro.
abstract: Nitric oxide (NO) has been shown to act as a potent antifibrogenic agent by decreasing myofibroblast differentiation. S-Nitroso-N-acetylcysteine (SNAC), a NO donor, attenuates liver fibrosis in rats, but the cellular and molecular mechanisms on liver myofibroblast-like phenotype still remain unknown. Here, we investigate the antifibrotic effects of SNAC on hepatic stellate cells, the major fibrogenic cell type in the liver. A murine GRX cell line was incubated with SNAC (100μM) or vehicle (control group) for 72h. Cell viability was measured by MTT colorimetric assay and the conversion of myofibroblast into quiescent fat-storing cell phenotype was evaluated by Oil-Red-O staining. TGFβ-1, TIMP-1, and MMP-13 levels were measure in the supernatant by ELISA. Profibrogenic- and fibrolytic-related gene expression was quantified using real-time qPCR. SNAC induced phenotype conversion of myofibroblast-like phenotype into quiescent cells. SNAC decreased gene and protein expression of TGFβ-1 and MMP-2 compared to control groups. Besides, SNAC down-regulated profibrogenic molecules and up-regulated MMP-13 gene expression, which plays a key role in the degradation of interstitial collagen in liver fibrosis. In conclusion, these findings demonstrate that SNAC efficiently can modulate the activation and functionality of murine hepatic stellate cells and could be considered as an antifibrotic treatment to human liver fibrosis.
authors.affiliation: Department of Gastroenterology, University of São Paulo School of Medicine, São Paulo, Brazil.
authors.forename: J T
authors.initials: JT
authors.lastname: Stefano
authors.forename: B
authors.initials: B
authors.lastname: Cogliati
authors.forename: F
authors.initials: F
authors.lastname: Santos
authors.forename: V M R
authors.initials: VM
authors.lastname: Lima
authors.forename: D C
authors.initials: DC
authors.lastname: Mazo
authors.forename: U
authors.initials: U
authors.lastname: Matte
authors.forename: M R
authors.initials: MR
authors.lastname: Alvares-da-Silva
authors.forename: T R
authors.initials: TR
authors.lastname: Silveira
authors.forename: F J
authors.initials: FJ
authors.lastname: Carrilho
authors.forename: C P M S
authors.initials: CP
authors.lastname: Oliveira
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D054337:Cell Dedifferentiation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D055166:Hepatic Stellate Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D008103:Liver Cirrhosis / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D051379:Mice
chemical_list: C024940:S-nitroso-N-acetylcysteine; D000111:Acetylcysteine