Chen Ying, Ji Lili, Xiong Aizheng, Yang Li, Wang Zhengtao
The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Toxicol Ind Health. 2013 Jul;29(6):567-75. doi: 10.1177/0748233712442707. Epub 2012 Apr 2.
Pyrrolizidine alkaloid isoline is isolated from the traditional Chinese medicine Ligularia duciformis. Our previous reports have already demonstrated isoline-induced liver injury in mice. The present study is designed to observe the involvement of intracellular reduced glutathione (GSH) in isoline-induced cytotoxicity in human normal liver L-02 cells. The results showed that isoline decreased the cellular GSH and the ratio of GSH and oxidized glutathione in a time- and concentration-dependent manner in L-02 cells. l-Buthionine-S-R-sulfoximine (BSO) is reported to inhibit cellular GSH biosynthesis, and further results showed that isoline decreased the cell viability in L-02 cells after pretreated with 25 μM BSO for 24 h. Furthermore, adducts of isoline and GSH were identified in L-02 cells using liquid chromatography/electrospray ionization tandem mass spectrometry (ion trap) for the first time. In conclusion, our study provides the strongest evidence to support the important roles of GSH in regulating isoline-induced cytotoxicity in human normal liver L-02 cells.
从传统中药双形橐吾中分离得到了吡咯里西啶生物碱异叶碱。我们之前的报告已经证明异叶碱可诱导小鼠肝损伤。本研究旨在观察细胞内还原型谷胱甘肽（GSH）在异叶碱诱导人正常肝L-02细胞毒性中的作用。结果表明，异叶碱在L-02细胞中以时间和浓度依赖性方式降低了细胞内GSH水平以及GSH与氧化型谷胱甘肽的比例。据报道，L-丁硫氨酸-S-R-亚砜胺（BSO）可抑制细胞内GSH的生物合成，进一步的结果显示，用25 μM BSO预处理24小时后，异叶碱降低了L-02细胞的活力。此外，首次使用液相色谱/电喷雾电离串联质谱（离子阱）在L-02细胞中鉴定出异叶碱与GSH的加合物。总之，我们的研究提供了最有力的证据，支持GSH在调节异叶碱诱导人正常肝L-02细胞毒性中发挥重要作用。
