TFEB Dysfunction Mediates Lysosomal Membrane Permeabilization-Induced Pyroptosis in Deoxynivalenol-Exposed Intestinal...

Deoxynivalenol (DON), a prevalent mycotoxin in cereals, triggers lysosomal membrane permeabilization (LMP) in intestinal epithelial cells, resulting in Transcription Factor EB (TFEB) dysfunction and subsequent pyroptosis. Both in vivo porcine jejunum and in vitro IPEC-J2 monolayer experiments revealed that DON exposure induced jejunal villus atrophy, tight junction disruption, mucin2 downregulation, cathepsin B/D leakage, and impaired TFEB nuclear translocation. LMP-mediated TFEB suppression activated NLRP3 inflammasome and gasdermin D (GSDMD)-dependent pore formation, culminating in caspase-1-driven pyroptosis. Pharmacological TFEB activation restored lysosomal integrity and attenuated pyroptosis, whereas TFEB knockdown exacerbated lysosomal instability. Mechanistically, TFEB maintained lysosomal homeostasis through LAMP1/LAMP2 upregulation. Notably, although NLRP3 inhibition by MCC950 mitigated pyroptotic markers, it failed to rescue lysosomal integrity in TFEB-deficient cells. Our findings establish TFEB as a pivotal regulator connecting lysosomal function to pyroptosis, revealing its therapeutic potential for DON-induced intestinal injury.