Diosmetin alleviates AFB1-induced endoplasmic reticulum stress, autophagy, and apoptosis via PI3K/AKT pathway in mice

Aflatoxin B1 (AFB1), a prevalent agricultural mycotoxin, represents a serious health hazard to humans and animals owing to its toxic effects. Diosmetin (DIOS), a naturally occurring flavonoid, has demonstrated potential hepatoprotective properties. This research seeks to investigate the mechanisms by which DIOS mitigates AFB1-induced hepatotoxicity in mice. The mice were divided into four groups: control (CON), AFB1, DIOS+AFB1, and DIOS. Over a 28 - day period, all groups were administered their respective treatments via oral gavage. The CON group was given an equivalent volume of PBS, the AFB1 group received AFB1 (0.4 mg/kg/day), the DIOS+AFB1 group was treated with DIOS (20 mg/kg/day) in combination with AFB1 (0.4 mg/kg/day), and the DIOS group received DIOS (20 mg/kg/day) alone. Then various experiments were used to evaluate the hepatotoxic effects of AFB1 and the hepatoprotective effects of DIOS in mice. Our findings initially demonstrated that AFB1 induced liver injury, oxidative stress, endoplasmic reticulum (ER) stress, apoptosis and autophagy. DIOS treatment notably ameliorated liver damage by lowering the LDH and MDA levels, increasing total antioxidant capacity and enhancing the GSH-Px, SOD and CAT activities. Additionally, DIOS dampened the secretion of inflammatory cytokines IL-1β and TNF-α, and blocked the NF-κB pathway. Moreover, DIOS administration lessened AFB1-induced ER stress-mediated apoptosis by inhibiting the mRNA and protein expressions of GRP78, p-PERK, p-elF2α, ATF6 and ATF4, while concurrently upregulating Bcl-2 expression and reducing the Bax and Cleaved Caspase-3 expressions. Furthermore, DIOS was also found to suppress the protein levels of LC3B, Beclin-1, ATG5, p62, and promote AKT phosphorylation. Overall, DIOS effectively mitigated AFB1-induced oxidative stress, inflammation, ER stress, apoptosis and autophagy via inhibition of the NF-κB pathway and stimulation of the PI3K/AKT pathway. The results imply that DIOS may be a viable therapeutic approach for the prevention of liver damage caused by AFB1 exposure.