龙葵碱诱导HepG2细胞凋亡的线粒体通路研究

 目的观察龙葵碱诱导HepG₂细胞凋亡与线粒体通路的关系。方法MTT法测定龙葵碱对HepG₂细胞的细胞毒作用;AO/EB双染,激光共聚焦扫描显微镜观察龙葵碱对HepG₂细胞形态学的影响;TMRE和Fluo-3/AM双染,激光共聚焦扫描显微镜同时观察细胞线粒体膜电位和细胞内Ca²⁺]_i的变化。结果龙葵碱对HepG₂细胞有细胞毒作用;龙葵碱诱导HepG₂细胞形态出现凋亡形态时TMRE和Fluo-3/AM双染观察表明,龙葵碱在降低线粒体膜电位的同时能够升高细胞内Ca²⁺]_i浓度。结论龙葵碱降低膜电位,开放细胞膜PT通道,使细胞内Ca²⁺顺浓度梯度转运,从而升高细胞内Ca²⁺浓度启动细胞凋亡机制。

Study on Mitochondrion Pathway of the Apoptosis of HepG2 Induced by Solanine

OBJECTIVE To investigate the relationship between the apoptosis of HepG₂ cells induced by solanine and the mitochondria PT channels.METHODS The cytotoxicity of solanine on HepG₂ cells was determined using MTT.The effect of solanine on the morphology of HepG₂ cells was observed using AO/EB double staining and LCSM.The changes in the mitochondrial membrane potential and intracellular Ca²⁺]_i was observed using TMRE,Fluo-3/AM double staining,and LCSM.RESULTS Solanine showed a cytotoxic effect on HepG₂ cells.When apoptosis appeared in HepG₂ cells under the induction of solanine,TRME and Fluo-3/AM double staining showed that solanine increased the concentration of intracellular Ca²⁺ and decreased the mitochondrial membrane potential.CONCLUSION Solanine can decreas membrane potential and open PT channels on cell membrane,thus get intracellular Ca²⁺ transport down its concentration gradient,which results in the increase of Ca²⁺ concentration and the initiation of the mechanism for cell apoptosis.