Effect of luteolin,levistilide A,and astragaloside Ⅰ on the proliferation and activation of hepatic stellate cell line LX-2 by single and combined administration
Objective To investigate the synergistic effect of luteolin(Lut),levistilide A(Lev),and astragaloside Ⅰ(Ast)on the proliferation and activation of human hepatic stellate cell line LX-2 by single and combined administvation.Methods LX-2 cells in the logarithmic growth phase were evaluated by methyl thiazolyl tetrazolium(MTT)method for the effects of Lut,Lev,and Ast on the proliferation of LX-2 cells induced by transforming growth factor-β1(TGF-β1),determination of combination ratio,and combination of Lut,Lev,and Ast on the proliferation of LX-2 cells induced by TGF-β1.The combination indexes(CI)of Lut,Lev,and Ast to inhibit the proliferation of LX-2 cells were calculated by CalcuSyn software to determine the types of combined use of the three.The effects of them on the protein expression of α-smooth actin(α-SMA)and TGF-β1 in LX-2 cells were evaluated by Western blot.Results The results of MTT experiments showed that the molar ratio of Lut,Lev,and Ast in combination was 1:1:10 and the single use of the and in combination of Lut,Lev,and Ast had inhibitory effects on the proliferation of LX-2 cells induced by TGF-β1(P＜0.05)in a concentration-dependent manner;CalcuSyn software analysis results displayed CI＜1 when the cell proliferation inhibition rate of LX-2 cells induced by TGF-β1 was more than 15％,indicating that combination of the three compounds was more synergistic and effective.Western blot results showed that the combined use of Lut,Lev,and Ast significantly reduced the expression of α-SMA protein in LX-2 cells induced by TGF-β1(P＜0.01)and the expression of TGF-β1 protein in LX-2 cells(P＜0.05).Conclusion Although the single use of each of Lut,Lev,and Ast can inhibit the proliferation and activation of LX-2 cells,the combination use of the three has stronger and more synergistic clinical result.It may be related to the down-regulation of TGF-β1 protein expressions in hepatic stellate cells.
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