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[Ca^2+]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[Ca^2+]i浓度。结论龙葵碱降低膜电位,开放细胞膜PTT通道,使细胞内Ca^2+顺浓度梯度转运,从而升高细胞内Ca^2+浓度启动细胞凋亡机制。
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 [ Ca^2+ ]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 Ca^2+ and decreased the mitochondrial membrane potential. CONCLUSION Solanine can decreas membrane potential and open PT channels on cell membrane, thus get intracellular Ca^2+ transport down its concentration gradient, which results in the increase of Ca^2+ concentration and the initiation of the mechanism for cell apoptosis.