MicroRNAs expression profiles in activation of hepatic stellate cells

Objective To determine the miRNAs expression profile of quiescent and culture activated rat hepatic stellate cells. Methods HSCs were isolated from the livers of adult Sprague-Dawley rats(weighing 400～500 g) by in situ perfusion and purified by single-step density gradient centrifugation with Nycodenz,became highly activated after 14 days' cultivation.Cell morphological characteristics were identified by means of phase-contrast microscopy,fluorescence microscopy. The difference in miRNAs expression between quiescent hepatic stellate cells at 2 days and their corresponding activated cells that underwent activation in vitro over a period of 2 weeks was detected by both microarrays and quantitative PCR. Results Twelve miRNAs were expressed above the threshold levels in activated hepatic stellate cells,with a progressively lower numbers when activated(P0.05).Only 9 miRNAs were expressed above the threshold levels in quiescent hepatic stellate cells.The expression of selected miRNAs was further validated and found their differential expression in cell-specific manners(P0.05). Conclusion The expression of a number of miRNAs in quiescent and activated hepatic stellate cells may suggest the potential roles of these miRNAs and applicability as diagnostic and prognostic signatures in liver fibrosis.