Apoptosis in HepaRG and HL-7702 cells inducted by polyphyllin II through caspases activation and cell-cycle arrest
Apoptosis in HepaRG and HL-7702 cells inducted by polyphyllin II through caspases activation and cell-cycle arrest
复制标题
DOI：
10.1002/jcp.27462
复制
发表时间：
2019-05-01
影响因子：
5.6
通讯作者：
Ni, Jian
中科院分区：
文献类型：
--
作者：
Wang, Wenping;Dong, Xiaoxv;Ni, Jian
传统中药Paragraphy在癌症预防和治疗方面显示出良好的前景。叶多酚II是麦冬中最重要的皂苷类成分之一，对多种肿瘤细胞具有一定的毒性作用。然而，我们在这项研究中的结果证明，polyphyllin II在体外具有肝毒性，通过半胱天冬酶激活和细胞周期阻滞。3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四氮唑结果表明，polyphyllin II抑制HepaRG和HL-7702细胞增殖，诱导细胞凋亡，并呈浓度和时间依赖性。然后，我们选择了创新的细胞模型-HepaRG细胞来探讨肝毒性的机制。我们的数据显示，活性氧（ROS）增加，线粒体膜电位降低后，给药polyphyllin II的HepaRG细胞。此外，随着浓度的增加，乳酸脱氢酶的释放增加，细胞周期的S期阻滞。而抗氧化剂N-乙酰半胱氨酸预处理后，HepaRG细胞凋亡明显减少，ROS产生减少，膜电位下降明显改善。此外，polyphyllin II处理增加Fas，Bax，细胞色素c，活化的caspase-3，-8，-9，裂解的聚（ADP-核糖）聚合酶的水平，并降低Bcl-2的表达水平。最后，我们确定了多叶素II诱导细胞凋亡的两条信号通路：死亡受体通路和线粒体通路。本研究证实了叶多酚II的体外肝毒性，这是迄今为止从未发现的，并为麦冬的临床应用敲响了警钟。
Rhizoma Paridis, a traditional Chinese medicine, has shown promise in cancer prevention and therapy. Polyphyllin II is one of the most significant saponins in Rhizoma Paridis and it has toxic effects on kinds of cancer cells. However, our results in this study proved that the polyphyllin II has hepatotoxicity in vitro through caspases activation and cell-cycle arrest. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide results indicated polyphyllin II inhibited proliferation, induced apoptosis in HepaRG cells and HL-7702 cells and showed a concentration and time-dependent. Then, we selected the innovative cell model-HepaRG cells to explore the mechanism of hepatotoxicity. Our data showed the reactive oxygen species (ROS) increased and the mitochondrial membrane potential decreased in HepaRG cells after administration of polyphyllin II. Besides, with the increase of concentration, the release of lactate dehydrogenase increased and the S phase of the cell cycle was arrested. Nevertheless, when pretreatment with antioxidant N-acetylcysteine, apoptotic cells decreased significantly, inhibited the production of ROS and improved the decrease of membrane potential in HepaRG cells. Moreover, polyphyllin II treatment increased levels of Fas, Bax, cytochrome c, activated caspase-3, -8, -9, cleaved poly(ADP-ribose) polymerase and decreased Bcl-2 expression levels. Finally, we identified two signal pathways of apoptosis induced by polyphyllin II including the death receptor pathway and the mitochondria pathway. This study confirmed the hepatotoxicity of the polyphyllin II in vitro, which has never been discovered and gave a wake-up call for the clinical application of Rhizoma Paridis.