Glyphosate induces intestinal inflammatory injury associated with mitochondrial-lysosomal dysfunction, cGAS-STING activation, and pyroptosis-related responses in common carp (Cyprinus carpio L.).

Glyphosate (GLY) poses a potential risk to fish health, yet the molecular mechanisms underlying GLY-induced intestinal injury remain unclear. This study established in vivo (common carp midgut) and in vitro (EPC cells) models to investigate the toxic effects of GLY. We evaluated ROS production, mitochondrial and lysosomal function, autophagic flux, cGAS-STING signaling, and pyroptosis via histology, fluorescence staining, RT-qPCR, and western blotting. Both in vivo and in vitro results showed that GLY exposure markedly increased ROS production, induced mitochondrial damage and lysosomal dysfunction, and impaired autophagic flux. GLY also promoted mitochondrial DNA (mtDNA) release into the cytoplasm, directly evidenced by increased cytosolic MT-ND1/MT-ND2 levels, and activated the cGAS-STING pathway. In vitro, pharmacological inhibition of STING with C-176 abrogated GLY-induced NLRP3 inflammasome activation, Caspase-1 cleavage, IL-1β maturation, GSDMD-N generation, and LDH release, supporting that STING acts upstream of the pyroptotic cascade. In summary, GLY exposure is associated with intestinal inflammatory injury in common carp, characterized by ROS overproduction and GSDMD-dependent pyroptosis. In vitro experiments in EPC cells provide mechanistic evidence for these pathways. This study offers new insights into GLY-induced intestinal toxicity and a theoretical basis for ecological risk assessment.