BAPTA-AM对HepG-2细胞MT的保护作用及其机制

Protective effects of BAPTA-AM on hydrogenperoxide-induced mitochondria injury in HepG-2 cells

目的观察BAPTA-AM抗H2O2诱导的HepG-2细胞MT损伤作用并探讨其机制。方法采用四甲基偶氮唑盐比色(MTT)、Rhodamine 123(Rh 123))荧光染色、细胞色素C酶联免疫分析、Caspase-9活性检测试剂盒、活性氧ROS检测法分别测定HepG-2细胞活性、线粒体(MT)膜电位、MT细胞色素C(CytC)的释放、Caspase-9的激活、活性氧(ROS)的产生量。结果 BAPTA-AM能抑制H2O2损伤HepG-2细胞所致MT跨膜电位的降低,减少CytC的释放,抑制Caspase-9激活和ROS的生成,提高受攻击H2O2的细胞活力。结论 BAPTA-AM通过减轻钙超载,保护MT,抑制凋亡通路的激活,产生抗细胞损伤、挽救细胞生命之效。

Aim This study was performed to investigate the protective effects of BAPTA-AM on hydrogen peroxide（H2O2）-induced mitochondria injury in HepG-2 cells and its possible mechanism.Methods HepG-2 cells were pretreated with BAPTA-AM liposome of different concentrations（10-8,10-7,10-6 and 10-5 mol·L-1） before 500 μmol·L-1 H2O2 treatment.The viabilities of HepG-2 cells（by MTT assay） were measured.Mitochondrial membrane potential（MMP）,cytochrome C,caspase-9 and reactive oxygen species（ROS） were tested to evaluate the inhibitory mechanisms.Results BAPTA-AM could significantly inhibit the collapse of MMP,the release of cytochrome C,the activation of caspase-9 and the elevation of ROS.Conclusion BAPTA-AM is a promising cell protector against H2O2-induced mitochondria injury,and its mechanisms are alleviating the calcium overload and suppression of the activation of apoptosis pathways.