Aflatoxin B1 (AFB1) exhibits hepatotoxic properties in both humans and animals. Contradictory findings regarding corticosterone suggest that it may either aggravate AFB1 toxicity or reduce its Lethal Dose 50 % (LD50), potentially through the role of the glucocorticoid receptor (GR). Additionally, microRNAs (miRNAs) are known to modulate the toxic effects of AFB1. Nevertheless, whether the modulation of GR-targeting miRNAs can alleviate AFB1-induced hepatotoxicity has not been thoroughly investigated. This study examined the expression of GR and its associated microRNAs in AFB1-induced hepatotoxicity in mice, using GR-targeting antagomirs to mitigate AFB1 toxicity. AFB1 exposure elicited liver inflammation and oxidative stress in mice, while also reducing detoxification capacity. Notably, a decrease in GR protein expression was observed in liver tissue and hepatocytes. Additionally, miR141-3p, miR200a-3p, miR384-5p, miR183-5p, miR181a-5p, and miR181b-5p were upregulated and identified as regulators of GR expression. AFB1 induced cytotoxicity in AML12 cells, as evidenced by decreased GR protein levels and increased expression of miR141-3p, miR200a-3p, and miR495-3p. Inhibition of miR141/200a/495-3p reduced AFB1-induced cytotoxicity in AML12 cells. Furthermore, GR-targeting antagomirs (antagomir141/200a/495-3p) alleviated AFB1-induced hepatotoxicity in mice. This study highlights potential therapeutic targets for AFB1-induced liver diseases and offers new insights into strategies to mitigate the harmful effects of aflatoxin exposure.