Gasdermin D-mediated microglial pyroptosis exacerbates neurotoxicity of aflatoxins B1 and M1 in mouse primary microglia and neuronal cultures.

ABSTRACT

Aflatoxin B1 (AFB1) disrupts the blood-brain barrier by poisoning the vascular endothelial cells and astrocytes that maintain it. It is important to examine whether aflatoxin B1 or its metabolite, aflatoxin M1 (AFM1), affect microglia, which as the "immune cells" in the brain may amplify their damaging effects. Here we evaluated the toxicity of AFB1 and AFM1 against primary microglia and found that both aflatoxins decreased the viability of primary microglia and increased the leakage of lactate dehydrogenase, gamma-H2AX expression, nuclear lysis, necrosis and apoptosis in a dose-dependent manner. The potential contribution of microglia to the toxic effects of aflatoxins was assessed in transwell co-culture experiments involving microglia, neurons, astrocytes, oligodendrocytes or neural stem/precursor cells. And we found that the toxic effects of both aflatoxins on various types of nervous system cells were greater in the presence of microglia than in their absence. We also found that both aflatoxins induced gasdermin D-mediated microglial pyroptosis and inflammatory cytokine expression by activating the NLRP3 inflammasome. Blockade of gasdermin D activity in AFB1- or AFM1-treated primary microglia using dimethyl fumarate (DMF) reduced the release of IL-1ß, IL-18 and nitric oxide, as well as the neurotoxicity of microglia-conditioned medium to neurons, astrocytes, oligodendrocytes and neural stem/precursor cells. These data suggested that the toxicity of AFB1 and AFM1 on various cells of the central nervous system is due, remarkably, the gasdermin D-mediated microglial pyroptosis exacerbates their neurotoxicity.