2. Academic Validation
  3. α-amanitin induces autophagy through AMPK-mTOR-ULK1 signaling pathway in hepatocytes

α-amanitin induces autophagy through AMPK-mTOR-ULK1 signaling pathway in hepatocytes

  • Toxicol Lett. 2023 Jun 15;S0378-4274(23)00204-7. doi: 10.1016/j.toxlet.2023.06.004.
Yue Xu 1,  Shangwen Wang 1,  Chi-Kwan Leung 2,  Hao Chen 1,  Chan Wang 1,  Huijie Zhang 1,  Shuwei Zhang 1,  Yi Tan 1,  Haowei Wang 1,  Lin Miao 1,  Yi Li 1,  Yizhen Huang 1,  Xiaoxing Zhang 1,  Genmeng Yang 3,  Ruilin Zhang 4,  Xiaofeng Zeng 5
Affiliations

Affiliations

  • 1 Department of Forensic Medicine, School of Forensic Medicine, Kunming Medical University, Kunming, China.
  • 2 School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong; CUHK-SDU Joint Laboratory of Reproductive Genetics, School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China.
  • 3 Department of Forensic Medicine, School of Forensic Medicine, Kunming Medical University, Kunming, China. Electronic address: [email protected].
  • 4 Department of Forensic Medicine, School of Forensic Medicine, Kunming Medical University, Kunming, China. Electronic address: [email protected].
  • 5 Department of Forensic Medicine, School of Forensic Medicine, Kunming Medical University, Kunming, China. Electronic address: [email protected].
PMID: 37329965 DOI: 10.1016/j.toxlet.2023.06.004
Abstract

Amanitin poisoning is one of the most life-threatening mushroom poisonings. α-Amanitin plays a key role in Amanita phalloides intoxication. α-Amanitin shows toxic effects on the liver. However, the mechanism by which α-amanitin induces liver injury has not been elucidated. Autophagy plays a crucial role in maintaining cellular homeostasis and is closely related to the occurrence of a variety of diseases. Studies have shown that Autophagy may play an important role in the process of α-amanitin-induced liver injury. However, the mechanism of α-amanitin-induced Autophagy remains unclear. Thus, this study aimed to explore the mechanisms of α-amanitin in inducing hepatotoxicity in Sprague Dawley (SD) rats and the normal human liver cell line L02 cells. The SD rats and L02 cells exposed to α-amanitin were observed to determine whether α-amanitin could induce the Autophagy of rat liver and L02 cells. The regulatory relationship between Autophagy and the AMPK-mTOR- ULK pathway by exposing the Autophagy agonist (rapamycin (RAPA)), Autophagy Inhibitor (3-methylademine (3-MA)), and AMPK Inhibitor (compound C) was also explored. Autophagy-related proteins and AMPK-mTOR-ULK pathway-related proteins were detected using Western blot. The results of the study indicated that exposure to different concentrations of α-amanitin led to morphological changes in liver cells and significantly elevated levels of ALT and AST in the serum of SD rats. Additionally, the expression levels of LC3-II, Beclin-1, ATG5, ATG7, AMPK, p-AMPK, mTOR, p-mTOR, and ULK1 were significantly increased in the rat liver. And we found that L02 cells exposed to 0.5μM α-amanitin for 6h significantly induced Autophagy and activated the AMPK-mTOR-ULK1 pathway. Pretreated with RAPA, 3-MA, and compound C for 1h, the expression levels of autophagy-related proteins and AMPK-mTOR-ULK pathway-related proteins significantly changed. Our results indicates that Autophagy and the AMPK-mTOR-ULK pathway are involved in the process of α-amanitin-induced liver injury. This study may foster the identification of actionable therapeutic targets for A. phalloides intoxication.

Keywords

AMPK-mTOR-ULK1 signaling pathway; L02 cells; autophagy; liver injury; α-amanitin.

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