Investigation of the Lipid-Lowering Effect of Vitamin C Through GSK-3β/β-Catenin Signaling in Zebrafish.

Vitamin C (VC) is an essential nutrient for most fish species because of the absence of L-gulonolactone oxidase in the bodies of fish. VC plays a significant role in maintaining the physiological functions and in improving the growth performance, immunity, and survival of fish. In this study, zebrafish ( Danio rerio ) were treated with 8.2, 509.6, and 1007.5 mg/kg VC diets for 2 weeks, and the muscle samples were collected for gene expression analysis and biochemical index analysis. The results indicated that 509.6 and 1007.5 mg/kg VC diets inhibited glycogen synthase kinase-3β (GSK-3β) expression and induced the expression of β-catenin in the muscle of zebrafish. The mRNA expression of CCAAT/enhancer-binding protein α (C/EBPα) and fatty acid synthase (FAS), FAS activity, and the content of glycerol and triglyceride (TG) were decreased in the muscle by 509.6 and 1007.5 mg/kg VC diets. In addition, GSK-3β RNA interference was observed in zebrafish fed with 8.2 and 1007.5 mg/kg VC diets. It was found that GSK-3β RNA interference induced the mRNA expression of β-catenin but decreased the mRNA expression of C/EBPα and FAS, FAS activity, as well as the content of glycerol and TG in the muscle of zebrafish. In ZF4 cells, the mRNA expression of GSK-3β, C/EBPα, and FAS was decreased, but β-catenin expression was increased by 0.1 and 0.5 mmol/L VC treatments in vitro . The glycerol and TG content, and FAS activity in ZF4 cells were decreased by 0.1 and 0.5 mmol/L VC treatments. Moreover, the result of western blot indicated that the protein expression level of GSK-3β was significantly decreased and that of β-catenin was significantly increased in ZF4 cells treated with 0.1 and 0.5 mmol/L VC. The results from in vivo and in vitro studies corroborated that VC exerted the lipid-lowering effect through GSK-3β/β-catenin signaling in zebrafish.