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

AIM:To investigate the effects of Chinese herb Yigan Decoction on proliferation and apoptosis of the hepatic stellate cells (HSC) in vitro. METHODS: The study in vitro was carried out in the culture of HSC lines. Various concentrations of Yigan Decoction were added and incubated. Cell proliferation was detected with MTT colorimetric assay. Cell apoptosis was detected by electron microscopy, flow cytometry and TUNEL. RESULTS: The proliferation of HSC was inhibited by Yigan Decoction, which depending on dose and time significantly. The HSC proliferation rates of groups at the end concentrations 144 and 72(g.L(-1)) were 21.62% and 40.54% respectively, significantly lower than that of normal control group(P【0.01). The HSC proliferation rates of groups at the end concentrations 36, 18 and 9(g.L(-1)) were 54.05%, 45.95% and 51.35% respectively, lower than that of control group (P【0.05). When the end concentration was 4.5 g.L(-1), the proliferation rate was 83.78%, which appeared no significant differences compared with control group. At the same concentrations of 18 g.L(-1), the inhibitory effects of Yigan Decoction at 24 h, 48 h and 72 h time point were observed, the effects were time-dependent, and reached a peak at 72 h. Meanwhile, it was showed that the inducing effects of Yigan Decoction on HSC apoptosis were dose-dependent and time-dependent. The apoptosis index(AI) was detected by TUNEL. After Yigan Decoction had been incubated for 48 h at the end concentration of 18 g.L(-1), the AI (14.5+/-3.1)% was significantly higher than that of control group (4.3+/-1.3)% (P【0.01). When visualized under transmission electron microscopy, some apoptotic stellate cells were found, i.e. dilated endoplasmic reticulum, irregular nuclei, chromatin condensation and heterochromatin ranked along inside of nuclear membrane. By flow cytometry detection, after HSC was treated with Yigan Decoction at different concentrations of 36, 18 and 9(g.L(-1)) for 48 h, AI (%) were 13.3+/-3.2, 10.7+/-2.7 and 10.1+/-2.5 respectively, which were significantly higher than that of control group(4.1+/-1.9) (P【0.01). At the same concentration of 18 g. L(-1) for 24h, 48 h and 72 h, AI (%) were 9.3+/-1.8,10.7+/-2.7 and 14.6+/-4.3 respectively, which were significantly higher than that of control group (P【0.01). CONCLUSION: Yigan Decoction could significantly inhibit HSC proliferation and increase the apoptosis index of HSC dose-dependently and time-dependently, which may be related to its mechanism of antifibrosis.

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 higher than that ofcontrol group(4. 1 ± 1.9) (P ＜ 0.01). At the sameconcentration of 18g@ L-1 for 24h, 48h and 72h, Al (%) were9.3± 1.8、10.7± 2.7 and 14.6±4.3 respectively, which weresignificantly higher than that of control group ( P＜ 0.01).CONCLUSION: Yigan Decoction could significantly inhibitHSC proliferation and increase the apoptosis index of HSCdosedependently and time-dependently, which may berelated to its mechanism of antifibrosis.