苦参素对D-氨基半乳糖苷诱导的肝细胞凋亡的干预研究

Effect of Oxymatrine on D-Galactosamine-induced Hepatocyte Apoptosis

[目的]研究氧化苦参碱对肝细胞凋亡的影响。[方法]采用D-氨基半乳糖苷(D-GalN)诱导人源HL-7702肝细胞发生凋亡,探索最佳的凋亡诱导参数,建立肝细胞凋亡模型,并运用建立的体外肝细胞凋亡模型,探讨氧化苦参碱的抗凋亡作用。[结果]D-GalN在60mmol/L浓度下与HL-7702肝细胞孵育8 h为最优建模条件,在此处理条件下经凝胶电泳可见细胞凋亡特征性的DNA ladder;且与对照组相比,HL-7702细胞处理后其ALT、AST、SOD、MDA、Caspases-3水平均显著升高(P<0.05)。[结论]D-GalN诱导的HL-7702肝细胞凋亡的发生机制可能与线粒体信号转导途径有关,而氧化苦参碱对D-GalN引起的肝细胞凋亡具有较强的干预作用,其抗凋亡机制可能与抑制Caspases-3、CytC、TNFα和TGF-β1等凋亡因子和诱导抗凋亡因子HO-1表达有关。

[Objective] To study effect of oxymatrine on d-galactosamine-induced hepatocyte apoptosis. [ Methods] The optimalizing parameters of d-gaiaetosamine （d-GaiN）-induced hepatoeyte apoptosis were determined and then the model of cell apoptosis induced by d-GaiN was estab- lished in human cell line HL-7702 hepatocytes. The anti-apoptotic effect of oxymatrine was evaluated by using the model established. [ Result] The incubation with 60 mmol/L d-GaiN for 8 hours may be the optimized conditions of hepatocyte apoptosis induced by d-GaiN. Typical apoptosis ladder was observed at 6 h after the HL-7702 hepatocytes were incubated with 60 mmol/L d-GaiN. Also the caspase-3, AST, ALT and MDA lev- els of the d-GaiN-sensitized hepatocytes were significantly elevated. [ Conclusion] HL-7702 hepatocyte apoptosis induced by d-GaiN is probably associated with the mitochondrial pathway. Additionally, oxymatrine exhibited significant inhibition on d-GaiN-induced hepatocyte apoptosis and its anti-apoptotic effect may contribute to the inhibition on apoptosis induce factors （ Caspases-3, CytC, TNFoL and TGF-β1 ） and induction of an- ti-aooototic cvtokine heme oxwenase-1 （HO-1）.