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Published on: November 27, 2019
α-amanitin induce inflammatory response by activating ROS/NF-κB-NLRP3 signaling pathway in human hepatoma HepG2 cells
Zhiyong Zhao1, Siliang Yi2, Hengchao E3
1Institute for Agro-food Standards and Testing Technology, Laboratory of Quality & Safety Risk Assessment for Agro-products (Shanghai), Ministry of Agriculture and Rural Affairs, Shanghai Academy of Agricultural Sciences, No.1000 Jinqi Road, Shanghai, 201403, PR China; Shanghai Guosen Biotechnology Co., Ltd., Shanghai, 201400, PR China.
Abstract:
α-amanitin (AMA) is a hepatotoxic mushroom toxin responsible for over 90% of mushroom poisoning fatalities worldwide, seriously endangering human life and health. Few evidences have indicated that AMA leads to inflammatory responses and inflammatory infiltration in vitro and in vivo. However, the molecular mechanism remains unknown. In this study, human hepatocellular carcinomas cells (HepG2) were exposed to AMA at various concentrations for short period of times. Results revealed that AMA increased ROS production and elevated the releases of malondialdehyde (MDA) and lactate dehydrogenase (LDH), resulting in oxidative damage in HepG2 cells. Also, AMA exposure significantly increased the secreted levels of inflammatory cytokines and activated the NLRP3 inflammasome. The inflammatory responses were reversed by NLRP3 inhibitor MCC950 and NF-κB inhibitor Bay11-7082. Additionally, N-acetylcysteine (NAC) blocked the upregulation of the NF-κB/NLRP3 signaling pathway and remarkably alleviated the inflammatory response. These results demonstrated that AMA could induce inflammation through activating the NLRP3 inflammasome triggered by ROS/NF-κB signaling pathway. Our research provides new insights into the molecular mechanism of AMA-induced inflammation damage and may contribute to establish new prevention strategies for AMA hepatotoxicity.
Insights
α-amanitin (AMA), a deadly mushroom toxin, triggers inflammation and oxidative damage in liver cells. This occurs via the ROS/NF-κB signaling pathway, activating the NLRP3 inflammasome, offering new prevention strategies for AMA hepatotoxicity.
Area of Science:
- Hepatology
- Toxicology
- Molecular Biology
Background:
- α-amanitin (AMA) is a potent hepatotoxic mushroom toxin responsible for most fatal mushroom poisonings.
- AMA's role in inducing inflammatory responses is known, but the underlying molecular mechanisms are unclear.
Purpose of the Study:
- To elucidate the molecular mechanism by which AMA induces inflammation and oxidative damage in liver cells.
- To investigate the involvement of reactive oxygen species (ROS), NF-κB, and the NLRP3 inflammasome in AMA-induced hepatotoxicity.
Main Methods:
- Human hepatocellular carcinoma cells (HepG2) were treated with varying concentrations of AMA.
- Assessed ROS production, malondialdehyde (MDA), and lactate dehydrogenase (LDH) release.
- Measured inflammatory cytokine levels and analyzed NLRP3 inflammasome activation.
- Utilized NLRP3 inhibitor (MCC950), NF-κB inhibitor (Bay11-7082), and antioxidant (N-acetylcysteine, NAC).
Main Results:
- AMA exposure increased ROS production, MDA, and LDH, indicating oxidative damage in HepG2 cells.
- AMA significantly elevated inflammatory cytokine secretion and activated the NLRP3 inflammasome.
- Inhibition of NLRP3 and NF-κB, as well as NAC treatment, reversed inflammatory responses and alleviated damage.
- NAC blocked the NF-κB/NLRP3 pathway activation.
Conclusions:
- AMA induces liver inflammation and oxidative damage by activating the NLRP3 inflammasome through the ROS/NF-κB signaling pathway.
- This study provides novel insights into AMA's molecular mechanism of toxicity.
- Findings may aid in developing new prevention and treatment strategies for α-amanitin poisoning.
