Effects of Yigan Decoction on proliferation and apoptosis of hepatic stellate cells

AIM: To investigate the effects of Chinese herb YiganDecoction on proliferation and apoptosis of the hepaticstellafe cells (HSC) in vitro.METHODS: The study in vitro was carried out in the cultureof HSC lines. Various concentrations of Yigan Decoctionwere added and incubated. Cell proliferation was detectedwith MTT colorimetric assay. Cell apoptosis was detected byelectron microscopy, flow cytometry and TUNEL.RESULTS: The proliferation of HSC was inhibited by YiganDecoction, which depending on dose and time significantly.The HSC proliferation rates ofgroups at the endconoentrations 144 and 72 (g@L-1 ) were 21.62 % and 140.54 %respectively, significantly lower then that of normal controlgroup( P ＜ 0.01 ). The HSC proliferation rates of groups atthe end concentratiors 36, 18 and 9(g@L-1 ) were 54.05 %,45.95 % and 51.35 % respectively, lower than that of controlgroup( P ＜ 0.05). When the end concentration was 4.5 g@L-1, the proliferation rate was 83.78 %, which appeared nosignificant differences compared with control group. At thesame concentrations of 18 g@L-1, the inhibitory effects ofYigan Decoction at 24 h, 48 h and 72 h time point wereobserved, the effects were time-dependent, and reached apeak at 72 h. Meanwhile, it was showed that the inducingeffects of Yigan Decoction on HSC apoptosis were dose-dependent and time-dependent. The apoptosis index (Al)was detected by TUNEL. After Yigan Decoction had beenincubated for 48 h at the end concentration of 18 g@ L-1 , tieAl (14.5 + 3. 1 ) % was significantly higher than that ofcontrol group (4.3+ 1.3) % (P＜0.01). When visualizedunder transnission electron microscopy, some apoptoticsfellafe cells were found, i. e. dilated endoplasmicreticulum, irregular ntclei, chromatin condensation andheterochromatin ranked along inside of nuclear membrane.By flow cytometry detection, after HSC was treated withYigan Decoction at different concentrations of 36, 18 and 9(g@ L-1 ) for 48 h, Al ( % ) were 13.3 ± 3.2, 10.7 ± 2.7 and 10.1 ±2.5 respectively, which were significantly