Astaxanthin inhibits proliferation and induces apoptosis of LX2 cells by regulating the miR29b/Bcl2 pathway

The aim of the present study was to investigate the role of microRNAs (miRNAs/miRs) in the antifibrotic effect of astaxanthin (AST), using the human hepatic stellate cell (HSC) line LX2 as the research model. LX2 cells were treated with various concentrations of AST (10, 20 and 40 µM) for 24 or 48 h. miR29b was selected based on existing literature, and its targeting gene B cell lymphoma (Bcl)2 was predicted by TargetScan and miRanda databases for further analysis. Interactions between miR29b and Bcl2 in the AST treated LX2 cells were evaluated using reverse transcriptionquantitative polymerase chain reaction (RTqPCR) and western blot analysis. MTT analysis was used to analyze cell viability. Overexpression of miR29b decreased the expression of Bcl2 in ASTtreated LX2 cells, and silencing of it had the opposite effect. Additionally, Annexin Vfluorescein isothiocyanate/propidium iodide double staining and flow cytometry were used to evaluate the cell apoptosis, and overexpression of miR29b increased cell apoptosis rates in ASTtreated LX2 cells; however, silencing of it had the opposite effect. RTqPCR and western blotting demonstrated that AST induced LX2 cells apoptosis which may be by regulating miR29b, as indicated by inhibited Bcl2 expression levels and elevated Bax and Caspase3 expression levels. These results highlight an important role of miR29b in the AST modulating LX2 cells proliferation and apoptosis and implicate a potential mechanism of miR29b and AST preventing liver fibrosis.