Metabolomic insights into clothianidin induced disorders of hepatic lipid metabolism in mice via AMPK signaling pathway

Clothianidin (CLO) is a second-generation neonicotinoid insecticide which is highly effective at killing insects by specifically activating their nicotinic acetylcholine receptors (nAChRs). Recent studies have shown that even low doses of CLO can cause behavioral abnormalities in mice and exhibit multisystemic toxic effects, including reproductive toxicity, neurotoxicity, and hepatotoxicity, further disrupting lipid metabolism homeostasis. However, the effects of CLO on hepatic lipid metabolism remain unclear at present. This study used male C57 mice as a model and administered CLO at four dose levels (0, 4, 16 and 64 mg/kg/day) via oral gavage for 14 consecutive days. Liver and serum samples from the control group and the group exposed to a high dose of CLO were analyzed using non-targeted metabolomics. The effects of CLO on liver lipid metabolism were evaluated using RT-qPCR, immunohistochemistry and Western blot techniques. Experimental results indicate that exposure to CLO causes significant liver damage and energy metabolism disorders. The mechanism of action is that CLO exposure causes energy deficiency, which in turn activates the AMPK signaling pathway to regulate energy metabolism. Specifically, this is manifested by a downregulation in the expression levels of key downstream proteins involved in lipid synthesis and an upregulation in the expression of proteins related to lipid degradation, ultimately leading to disorders in hepatic lipid metabolism. In summary, this study elucidates how CLO exposure disrupts energy metabolism and mediates lipid metabolism disorders through the AMPK signaling pathway, while also providing new theoretical basis for assessing the toxicological effects of CLO.