The endoplasmic reticulum-mitochondrial crosstalk is involved in the mitigation mechanism of eucalyptol on imidacloprid toxicity in Ctenopharyngodon idellus kidney cells.

Imidacloprid (IMI), a systemic neonicotinoid insecticide widely used in agriculture, resulting in persistence in aquatic environments that threaten the survival of organisms. Eucalyptol (EUC), a monoterpenoid found in plants, can be applied to medicine, food, and aquaculture. However, the potential protective effects of EUC on cell damage under neonicotinoid pesticide toxicity, and the role of ER stress and its mediated apoptosis and necroptosis in it, remain unclear. Therefore, we treated Ctenopharyngodon idellus kidney (CIK) cells with 20 mg/L IMI and 20 µM EUC for 48 h. The results showed that IMI exposure caused a higher GRP78 levels, activated ATF6, PERK-eIF2α and IRE1-XBP1 pathways, led to the decline of ATPase activities and ATP content, induced the expression of cytokine (TNF-α, IL-1ß, IL-6 and INF-γ), triggered BCL2/BAX-mediated apoptosis and RIP1/RIP3/MLKL-dependent necroptosis in the CIK cell line. Surprisingly, EUC had an effect against IMI-induced cytotoxicity, showing that it effectively mitigated the above-mentioned IMI-exposure-induced changes. Taken together, these results suggested that EUC could alleviated IMI-induced cell death and dysimmunity by recovering ER stress/mitochondria imbalance. These results partly explained the mechanism of biological threat on fish under IMI exposure and the potential application value of EUC in aquaculture.