苦参碱与氧化苦参碱抑制HepG2细胞增殖的对比研究

目的比较研究苦参碱与氧化苦参碱体外抑制肝癌HepG2细胞增殖的效果并初步探讨其机制。方法不同剂量苦参碱与氧化苦参碱注射液处理细胞 ,细胞计数及溴化四唑蓝实验观察细胞增殖抑制 ;光镜观察细胞形态改变 ;流式细胞仪和Tunnel实验观察细胞凋亡并检测细胞周期 ;免疫组化观察凋亡相关基因Bax与Bcl 2的表达。结果 0 .8g/L苦参碱处理细胞 3d即可明显抑制HepG2细胞增殖 ,细胞存活率为 5 1% ;1.5g/L苦参碱可明显诱导细胞凋亡 ,用药后大量细胞被阻滞于S期 ;1.5g/L的苦参碱使HepG2细胞凋亡相关基因Bax表达增强 ,Bcl 2的表达减弱。与苦参碱同浓度的氧化苦参碱不能抑制细胞增殖 ,1.5g/L氧化苦参碱处理细胞 3d ,细胞存活率仍为84 .2 % ;氧化苦参碱的浓度增至 8.0g/L时才出现明显凋亡峰。结论体外苦参碱诱导HepG2细胞凋亡能力强于氧化苦参碱 ,并调控了凋亡相关基因的表达 ,而氧化苦参碱浓度 6倍于苦参碱时细胞才出现凋亡。

Comparative study on inhibiting proliferation of HepG2 cells by matrine and oxmatrine

PurposeTo observe the differences of the effects and the possible mechanisms in the inducing apoptosis and inhibiting proliferation by matrine and oxmatrine in HepG2 cells.MethodsHepG2 cells were treated by different doses of matrine and oxmatrine. The suppressive effect of matrine on the proliferation of HepG2 cells was observed by MTT test and cell counts. The apoptosis and cell cycle were observed by flow cytometric assay and TUNEL in situ assay. The expression of Bcl-2 and Bax were investigated with immunohistochemical analysis.Resultsin the concentration of 0.8 g/L could inhibit the proliferation of HepG2 cells (the 3-day survival rate was 51%).Matrine in the concentration of 1.5 g/L could induce apoptosis obviously. The expression of Bax gene was increased and that of Bcl-2 gene was suppressed. Most of the cells were blocked in the S phage. On the other hand, Oxmatrine could not inhibit the proliferation of the HepG2 cells at the same doses. The 1.5 g/L for 3-day cell survival rate was 84.2%. The apoptosis peak of cells did not appear until the dose of oxmatrine was up to 8.0 g/L.ConclusionThe effect of matrine inducing apoptosis and inhibiting proliferation of HepG2 cells in vitro is stronger than that of oxmatrine.It regulates the expression of gene associated with the apoptosis and when the dose of oxmatrine was 6 times more than that of matrine, apoptosis peak appears in the cells.