Liang Yonghong, Chen Sixin, Han Suqin, Luo Li, Shen Feihai, Huang Zhiying
School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, PR China.
School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, PR China; School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, PR China.
Toxicol Lett. 2023 Mar 15;377:51-61. doi: 10.1016/j.toxlet.2023.02.006. Epub 2023 Feb 16.
Toosendanin (TSN) is the main active compound of Melia toosendan Sieb et Zucc with various bioactivities. In this study, we investigated the role of ferroptosis in TSN-induced hepatotoxicity. The characteristic indicators of ferroptosis were detected including reactive oxygen species (ROS), lipid-ROS, glutathione (GSH), ferrous ion and the expression of glutathione peroxidase 4 (GPX4), which showed that TSN caused ferroptosis in hepatocytes. The results of qPCR analysis and western blotting assay showed that TSN-induced activation of protein kinase R-like endoplasmic reticulum kinase (PERK)- eukaryotic initiation factor 2 α subunit (eIF2α)- activation transcription factor 4 (ATF4) signaling pathway resulted in increasing activation transcription factor 3 (ATF3) expression, which upregulated the expression of transferrin receptor 1 (TFRC). Furthermore, TFRC mediated iron accumulation leading to ferroptosis in hepatocytes. To clarify whether TSN triggered ferroptosis in vivo, male Balb/c mice were treated with the different doses of TSN. The results of hematoxylin-eosin (H&E) staining, 4-hydroxynonenal (4-HNE) staining, malondialdehyde (MDA) content and the protein expression of GPX4 showed that ferroptosis contributed to TSN-induced hepatotoxicity. Iron homeostasis relative protein and PERK- eIF2α- ATF4 signaling pathway also involved in hepatotoxicity of TSN in vivo.
川楝素（TSN）是川楝（Melia toosendan Sieb et Zucc）的主要活性化合物，具有多种生物活性。在本研究中，我们探讨了铁死亡在TSN诱导的肝毒性中的作用。检测了铁死亡的特征指标，包括活性氧（ROS）、脂质ROS、谷胱甘肽（GSH）、亚铁离子以及谷胱甘肽过氧化物酶4（GPX4）的表达，结果表明TSN可导致肝细胞发生铁死亡。qPCR分析和蛋白质印迹检测结果显示，TSN诱导蛋白激酶R样内质网激酶（PERK）-真核起始因子2α亚基（eIF2α）-激活转录因子4（ATF4）信号通路活化，导致激活转录因子3（ATF3）表达增加，进而上调转铁蛋白受体1（TFRC）的表达。此外，TFRC介导铁蓄积，导致肝细胞发生铁死亡。为明确TSN在体内是否引发铁死亡，对雄性Balb/c小鼠给予不同剂量的TSN进行处理。苏木精-伊红（H&E）染色、4-羟基壬烯醛（4-HNE）染色、丙二醛（MDA）含量及GPX4蛋白表达结果显示，铁死亡参与了TSN诱导的肝毒性。铁稳态相关蛋白以及PERK-eIF2α-ATF4信号通路也参与了TSN在体内的肝毒性作用。
