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Aim A novel amphipathic curcumin derivative(Curc-OEG)that inchdes two short hydrophilic ethylene glycol oligomer(OEG)chains attached to the basic curcumin structure showed improved bioavailability and possessed anti-fibrotic activity in carbon tetrachloride-induced liver fibrosis in vivo in our previous work.However,the mechanisms underlying the anti-fibrotic effect of Curc-OEG remain unclear and were investigated in this study using primary and activated HSCs in vitro.Methods The fieshly isolated and activated HSCs as model systems were employed.HSCs were treated with different concentrations of Curc-OEG at 0,3.125,6.25,12.5,25.0,50,75 μg/ml.Cell morphology changes were visualized using a microscope.Apoptotic cells were analyzed by flow cytometry.The genes related to fibrogenesis and apoptosis were evaluated by real time PCR and Western blotting.Results Curc-OEG significantly inhibited the activation of primary HSCs and caused a significant concentration-dependent reduction in cell numbers of 55 and 85% at 6.25 and 12.5 μg/ml respectively compared with the control cells after 7d cultivation,while primary HSCs couldnt activate at high concentrations.At 12.5 μg/ml,Curc-OEG reduced mRNA levels of α-SMA,collagen I,TGF-β1 and Smad2 by 83,77,85 and 75%,respectively.In accordance with mRNA expression levels,Curc-OEG also reduced the levels of α-SMA,collagen I,TGF-β1 and Smad2 proteins by 94,82,92 and 73%,respectively.The apoptotic tate of activated HSCs significantly increased from about 6.0 to 99.8% upon treatment of Curc-OEG from 6.25 to 75 μg/ml for 24th.Moreover,Curc-OEG suppressed the mRNA levels of Bcl-2,α-SMA,collagens Ⅰ and ⅡⅢ,TGF-β1,Smad2,Smad3,PDGF-βR,NF-κB,TIMP-1,TIMP-2,and upregulated the mRNA expresion levels of Bax and PPAR-γ.Conclusion Curc-OEG targeted and interfered with the activation and proliferation of HSCs through multiple signalling pathways,and may be a promising drug candidate for treatment of liver fibrosis.