Gao Shi-Yong, Wang Qiu-Juan, Ji Yu-Bin
Department of Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu Province.
World J Gastroenterol. 2006 Jun 7;12(21):3359-67. doi: 10.3748/wjg.v12.i21.3359.
To observe the effect of solanine on the membrane potential of mitochondria in HepG(2) cells and Ca(2+) in the cells, and to uncover the mechanism by which solanine induces apoptosis.
HepG(2) cells were double stained with AO/EB, and morphological changes of the cells were observed using laser confocal scanning microscopy (LCSM). HepG(2) cells were stained with TMRE, and change in the membrane potential of mitochondria in the cells were observed using LCSM. HepG(2) cells were double stained with Fluo-3/AM, and change of Ca(2+) in the cells were observed using LCSM. HepG(2) cells were double stained with TMRE and Fluo-3/AM, and both the change in membrane potential of mitochondria and that of Ca(2+) in the cells were observed using LCSM.
Cells in treated groups showed typical signs of apoptosis. Staining with TMRE showed that solanine could lower membrane potential; staining with Fluo-3/AM showed that solanine could increase the concentration of Ca(2+) in tumor cells; and those of double staining with TMRE and Fluo-3/AM showed that solanine could increase the concentration of Ca(2+) in the cells at the same time as it lowered the membrane potential of mitochondria.
Solanine opens up the PT channels in the membrane by lowering the membrane po-tential, leading to Ca(2+) being transported down its concentration gradient, which in turn leads to the rise of the concentration of Ca(2+) in the cell, turning on the mechanism for apoptosis.
观察茄碱对HepG(2)细胞线粒体膜电位及细胞内[Ca(2+)]i的影响，揭示茄碱诱导细胞凋亡的机制。
采用AO/EB对HepG(2)细胞进行双重染色，利用激光共聚焦扫描显微镜（LCSM）观察细胞形态变化。用TMRE对HepG(2)细胞进行染色，利用LCSM观察细胞线粒体膜电位的变化。用Fluo-3/AM对HepG(2)细胞进行双重染色，利用LCSM观察细胞内[Ca(2+)]i的变化。用TMRE和Fluo-3/AM对HepG(2)细胞进行双重染色，利用LCSM观察细胞线粒体膜电位及[Ca(2+)]i的变化。
处理组细胞呈现典型的凋亡特征。TMRE染色显示茄碱可降低膜电位；Fluo-3/AM染色显示茄碱可增加肿瘤细胞内Ca(2+)浓度；TMRE和Fluo-3/AM双重染色显示茄碱在降低线粒体膜电位的同时可增加细胞内Ca(2+)浓度。
茄碱通过降低膜电位开放膜上的PT通道，导致Ca(2+)顺浓度梯度转运，进而使细胞内Ca(2+)浓度升高，启动细胞凋亡机制。
