Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China.
College of Chemical Engineering, Zhejiang University of Technology, Hangzhou 310008, China.
Toxins (Basel). 2022 Sep 13;14(9):633. doi: 10.3390/toxins14090633.
Pyrrolizidine alkaloids (PAs) are common constituents of plants and have serious hepatotoxicity. Intermedine (Im) and lycopsamine (La) are two monoesters of PAs that frequently coexist in the PA-containing plants (e.g., and tea). The present study aimed to explore the combined hepatotoxicity and toxicity mechanism of the Im and La mixture. In vitro, the combined cytotoxicity of the Im and La mixture on human hepatocytes (HepD) was examined by CCK-8, colony formation, wound healing, and Annexin V/PI staining assays. The combination of Im and La inhibited the ability of HepD cells to proliferate, colonize, and migrate and induced hepatocytes apoptosis in a dose-dependent manner. In addition to significantly causing a burst of intracellular reactive oxygen species (ROS), mitochondrial apoptosis, and endoplasmic reticulum (ER) stress, the Im and La mixture can also cause an increase in intracellular Ca, triggering the PERK/eIF2α/ATF4/CHOP apoptosis pathway. This study provided the first direct evidence that the combined PAs induced hepatotoxicity through ER-mediated apoptosis. These results supplemented the basic toxicity data for the combined PAs and provided a new perspective for the risk assessment of combined PA toxicity.
吡咯里西啶生物碱（PAs）是植物中的常见成分，具有严重的肝毒性。千里光宁（Im）和千里光裂碱（La）是 PAs 的两种单酯，它们经常共存于含有 PAs 的植物中（例如，和茶）。本研究旨在探讨 Im 和 La 混合物的联合肝毒性及其毒性机制。体外，通过 CCK-8、集落形成、划痕愈合和 Annexin V/PI 染色实验，研究了 Im 和 La 混合物对人肝细胞（HepD）的联合细胞毒性。Im 和 La 的混合物以剂量依赖性方式抑制 HepD 细胞增殖、定植和迁移的能力，并诱导肝细胞凋亡。除了显着引起细胞内活性氧（ROS）、线粒体凋亡和内质网（ER）应激的爆发外，Im 和 La 混合物还可以导致细胞内 Ca2+增加，触发 PERK/eIF2α/ATF4/CHOP 凋亡途径。这项研究首次直接证明了联合 PAs 通过 ER 介导的凋亡诱导肝毒性。这些结果补充了联合 PAs 的基本毒性数据，并为联合 PA 毒性的风险评估提供了新的视角。
