New insights into Microalgal astaxanthin's effect on Lambda-cyhalothrin-induced lymphocytes immunotoxicity in Cyprinus carpio: Involving miRNA-194-5p-FoxO1-mediated-mitophagy and pyroptosis.

Lambda-cyhalothrin (LC), a pyrethroid insecticide widely used in agriculture, causes immunotoxicity to aquatic organisms in the aquatic environment. Microalgal astaxanthin (MA) is a natural carotenoid that enhances viability of a variety of fish. To investigate the immunotoxicity of LC and the improvement effect of MA in lymphocytes (Cyprinus carpio), lymphocytes were treated with LC (80 M) and/or MA (50 M) for 24 h. Firstly, CCK8 combined with PI staining results showed that MA significantly attenuated the LC-induced lymphocyte death rate. Secondly, LC exposure resulted in excessively damaged mitochondrial and mtROS, diminished mitochondrial membrane potential and ATP content, which could be improved by MA. Thirdly, MA upregulated the levels of mitophagy-related regulatory factors (Beclin1, LC3, ATG5, Tom20 and Lamp2) induced by LC. Importantly, MA decreased the levels of pyroptosis-related genes treated with LC, including NLRP3, Cas-4, GSDMD and active Cas-1. Further study indicated that LC treatment caused excessive miRNA-194-5p and reduced levels of FoxO1, PINK1 and Parkin, which was inhibited by MA treatment. Overall, we concluded that MA could enhance damaged mitochondrial elimination by promoting the miRNA-194-5p-FoxO1-PINK1/Parkin-mitophagy in lymphocytes, which reduced mtROS accumulation and alleviated pyroptosis. It offers insights into the importance of MA application in aquaculture as well as the defense of farmed fish against agrobiological hazards in fish under LC.