Decabromodiphenyl ethane (DBDPE) and polystyrene nanoplastics (PS -NPs) are emerging pollutants that seriously threaten the ecological safety of the aquatic environment. However, the hepatotoxicity effect of their combined exposure on aquatic organisms has not been reported to date. In, this study, the effects of single or coexposure of DBDPE and PS -NPs on grass carp hepatocytes were explored and biomarkers related to oxidative stress, ferroptosis, and inflammatory cytokines were evaluated. The results show that both single and coexposure to DBDPE and PS -NPs caused oxidative stress. Oxidative stress was induced by increasing the contents of pro -oxidation factors (ROS, MDA, and LPO), inhibiting the activity of antioxidant enzymes (CAT, GPX, TSOD, GSH, and T-AOC), and downregulating the mRNA expressions of antioxidant genes ( GPX1 , GSTO1 , SOD1 , and CAT ); the effects of combined exposure were stronger overall. Both single and co -exposure to DBDPE and PSNPs also elevated Fe 2 + content, promoted the expressions of TFR1 , STEAP3 , and NCOA4 , and inhibited the expressions of FTH1 , SLC7A11 , GCLC , GSS , and GPX4 ; these effects resulted in iron overload -induced ferroptosis, where co -exposure had stronger adverse effects on ferroptosis-related biomarkers than single exposure. Moreover, single or co -exposure enhanced inflammatory cytokine levels, as evidenced by increased mRNA expressions of IL -6 , IL -12 , IL -17 , IL -18 , IL-1 beta , TNF- alpha , IFN- gamma , and MPO . Co -exposure exhibited higher expression of proinflammatory cytokines compared to single exposure. Interestingly, the ferroptosis inhibitor ferrostatin-1 intervention diminished the above changes. In brief, the results suggest that DBDPE and PS -NPs trigger elevated levels of inflammatory cytokines in grass crap hepatocytes. This elevation is achieved via oxidative stress and iron overload -mediated ferroptosis, where cytotoxicity was stronger under co -exposure compared to single exposure. Overall, the findings contribute to elucidating the potential hepatotoxicity mechanisms in aquatic organisms caused by co -exposure to DBDPE and PS -NPs.
