[Impact of artesunate on the expression and secretion of transforming growth factor-b1 of primary rat hepatic stellate cells].

Objective To investigate the impact of the Artemisia annua plant-derived drug,artesunate,on proliferation of primary rat hepatic stellate cells (HSCs),and to analyze the underlying molecular mechanisms of its anti-fibrogenic effects involving the inhibition of transforming growth factor-beta 1 (TGF-β1) expression and secretion in liver.Method Isolated,cultured,and activated primary rat HSCs were divided into sixteen groups,including one untreated control group and fifteen artesunate-treated experimental groups with 125,150,175,200 or 225 μmol/L for 24,48 or 72 hours.The rate of cellular proliferation was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.TGF-β1mRNA expression was evaluated by reverse transcription-polymerase chain reaction and protein expression was evaluated by Western blotting.Enzyme-linked immunosorbent assay was used to evaluate secreted levels of TGF-β1 protein.Results Artesunate significantly inhibited proliferation of cultured HSCs in a doseand time-dependent manner (all,P ＜ 0.01).Atter 24 hours of exposure,the inhibition ratios of the various artesunate concentrations were:6.06％± 1.44％ (125 μmol/L),21.47％±5.57％ (150 μmol/L),42.00％± 7.36％(175 μmol/L),67.12％ ± 4.55％ (200 μmol/L),and 79.83％ ±3.67％ (225 μmol/L).Artesunate significantly inhibited the TGF-β1 mRNA expression in HSCs,and the higher the drug concentration,the higher the degree of inhibition (all,P ＜ 0.01).In addition,artesunate significantly inhibited the expression of intracellular and secreted TGF-β1 protein (all,P ＜ 0.01).In response to artesunate (μmol/L concentrations),the TGF-β1 levels were (164.24±6.88) pg/ml (0μmol/L),(102.68±4.45) pg/ml (150μmol/L),(86.54±5.56) pg/ml (1751μmol/L),and (56.55 ± 5.66) pg/ml (200μmol/L).Conclusion Artesunate exerts anti-fibrogenic effects on HSCs in vitro,possibly by reducing the expression,translation and secretion of TGF-β1. Key words: Artemisia annua;  Liver cirrhosis;  Transforming growth factor beta;  Artesunate; Hepatic stellate cells