翌圣生物 Yeasen｜生命科学试剂与诊断原料一站式平台

Abstract

Ammonia nitrogen is a common pollutant in aquaculture water, which can induce oxidative stress and inhibit growth in fish. α-ketoglutarate (AKG) has been proven to play an important role in alleviating oxidative stress and maintaining mitochondrial homeostasis. To investigate the regulatory mechanism of α-ketoglutarate in ammonium chloride-induced hepatocyte ferroptosis, flow cytometry and Western blotting were performed with grass carp hepatocytes as the experimental model. The results showed that ammonium chloride exposure could significantly reduce the viability of grass carp L8824 hepatocytes, significantly increase the expression levels of lipid reactive oxygen species (LipROS) and ACSL4 protein, and simultaneously cause damage to the morphological structure and physiological function of mitochondria, thereby inducing lipid peroxidation. In addition, ammonium chloride could also disrupt the cellular iron ion transport system, leading to abnormal accumulation of intracellular iron ions and ultimately inducing hepatocyte ferroptosis. After AKG intervention, hepatocyte viability was significantly improved, the expression levels of LipROS was significantly decreased, and lipid peroxidation damage was effectively alleviated. Meanwhile, mitochondrial homeostasis was maintained but the expression levels of ferroptosis-related genes did not show a significant changes. In summary, AKG may alleviate ammonium chloride stress-mediated ferroptosis in grass carp L8824 hepatocytes by scavenging intracellular reactive oxygen species and repairing mitochondrial function. The results of this study provide a new idea for ammonia nitrogen pollution control and healthy fish culture in aquaculture.