Aim: The purpose of the current work was to demonstrate and compare the impacts of folate, methionine, and choline as methyl donors in the prevention of NAFLD induced by a high-fructose diet (HFrD). Material and Methods: Sixty adult male albino rats were divided into five groups: G1 had a control-fed basal diet; in G2, the rats were fed an HFrD containing 45% fructose (HFrD); in G3, G4, and G5, the rats were fed an HFrD supplemented with folic acid, L-methionine, and choline chloride, respectively. The present study's findings showed a close connection between methyl donors and lipid metabolism. Results: The findings indicated that the folic acid supplemented diet (HFrD+ folate) exhibited the most substantial improvement in hepatic total lipids (TP), triacylglycerol (TG), phospholipid (PL), serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), free fatty acid (FFA), albumin (ALB), total bile acid (TBA), fasting blood glucose (FBG), proinflammatory cytokines levels, and enzyme activities of alanine transaminase (ALT), aspartate transaminase (AST), fatty acid synthase (FAS), and acetyl coenzyme A carboxylase (ACC). In the same context, L-methionine had the most significant improvement in oxidative stress biomarker (glutathione [GSH], superoxide dismutase [SOD], 4-hydroxynonenal [4-HNE]) levels, followed by folic acid and choline. Discussion: Methyl donors such as folic acid, methionine, and choline are essential for maintaining OCM and can prevent animals with NAFLD from developing liver lipid accumulation.