Aflatoxin B1-induced liver pyroptosis is mediated by disturbing the gut microbial metabolites: the roles of pipecolic acid and norepinephrine

The disturbed gut microbiota is a key factor in activating the aflatoxin B1 (AFB1)-induced liver pyroptosis by promoting inflammatory hepatic injury; however, the pathogen associated molecular pattern (PAMP) from disturbed gut microbiota and its mechanism in activating liver pyroptosis remain undefined. By transplanting AFB1-originated fecal microbiota and sterile fecal microbial metabolites filtrate, we determined the association of PAMP in AFB1-induced liver pyroptosis. Notably, AFB1-originated sterile fecal microbial metabolites filtrate were more active in triggering liver pyroptosis in mice, as compared to parental fecal microbiota. This result supported a critical role of the metabolic homeostasis of gut microbiota in AFB1-induced liver pyroptosis, rather than an injurious response to direct exposure of AFB1 in liver. Among the gut-microbial metabolites, pipecolic acid and norepinephrine were proposed to bind TLR4 and NLRP3, the upstream proteins of pyroptosis signaling pathway. Besides, the activations of TLR4 and NLRP3 were linearly correlated with the concentrations of pipecolic acid and norepinephrine in the serum of mice. In silenced expression of TLR4 and NLRP3 in HepG2 cells, pipecolic acid or norepinephrine did not able to activate hepatocyte pyroptosis. These results demonstrated the necessity of gut microbial metabolism in sustaining liver homeostasis, as well as the potential to provide new insights into targeted intervention for AFB1 hepatotoxicity. As a common mycotoxin, Aflatoxin B1 (AFB1) is widely present in food and feed. It has the potential to negatively impact the stability and functioning of ecosystems, and may lead to bioaccumulation and biomagnification. To mitigate the impact of AFB1 on animal and human health, it is important to strengthen research on the mechanism of AFB1 action, and implement effective management and control strategies. This study explored the liver injury mechanism of AFB1 in depth, which opened a new idea for the prevention and treatment of hepatotoxicity of mycotoxins by regulating gut microbiota.