Inhibition effect of garlic bioactive component diallyltrisulfide on hepatic stellate cell activation induced by oxidative stress / 中国药学杂志

ABSTRACT

OBJECTIVE: To investigate the effects of the primary bioactive component diallyltrisulfide (DATS) contained in garlic on the activation of hepatic stellate cells (HSCs) induced by oxidative stress in vitro. METHODS: Rat HSCs were cultured in vitro, and MTT assay was used to screen the effective dose of hydrogen peroxide (H2O2) for stimulating HSC proliferation. Then lactate dehydrogenase assay was used to examine the toxic effect of DATS on HSCs, and MTT assay to evaluate the anti-proliferative effects of DATS at non-toxic concentrations on HSCs. Flow cytometry was used to determine DATS effects on cell cycle. Hoechst33258 staining and flow cytometry were used to analyze apoptosis. Western blot assays were used to examine the expression of relevant proteins and fibrotic markers. RESULTS: H2O2 at 5 μmol · L significantly promoted HSC proliferation, and this concentration was selected to establish the oxidative stress-induced HSC activation model. MTT assay showed that DATS dose-dependently inhibited HSC proliferation in the presence of H2O2. DATS arrested the H2O2-treated HSC at G2/M check point associated with downregulation of Cyclin B1 and CDK1. DATS also dose-dependently stimulated H2O2-treated HSCs to undergo apoptosis, which was associated with modulation of apoptosis-related molecules including Bax, Bcl-2, Caspase-9 and Caspase-3. Furthermore, DATS was found to reduce the protein abundance of a series of fibrotic marker proteins. CONCLUSION: DATS effectively inhibits oxidative stress-induced HSC activation in vitro demonstrated by suppressed proliferation, arrests cell cycle, increases apoptosis and reduces fibrotic marker expression. These findings provide novel insights into DATS as a potential antifibrotic candidate for further development.