Background: The Helichrysum genus of the Asteraceae family comprises 500 species. Fifteen Helichrysum species are found in Turkey. This genus is used in traditional medicine for several beneficial effects.Objectives: This study explored Helichrysum plicatum subsp. plicatum (HPsP) effects on fructose-induced nonalcoholic fatty liver disease (NAFLD). Methods: Thirty-five adult male Wistar rats weighing 200–250 g were divided into 5 groups: Control, HPsP, fructose (F), HPsP+fructose, and double-dose HPsP+fructose. Fructose (40%) was then added to the water. HPsP extract was administered at doses of 4 and 8 mg/kg. All treatments were performed for 8 weeks.Results: The F, F+HPsP, and double-dose groups exhibited higher palmitic acid (16:0) levels than the control group (P<0.001). F+HPsP treatment enhanced palmitoleic acid levels (P<0.001). The F and F+HPsP groups showed reduced levels of linolenic (18:3 n-3), eicosadienoic (20:2, n-6), and arachidonic (20:4 n-6) acids (P<0.001, P<0.01, and P<0.05, respectively). The F, F+HPsP, and double-dose groups showed lower docosahexaenoic acid (22:6, n-3) levels (P<0.05). Retinol and in HPsP treated group and (P<0.05) K1 vitamin levels increased in all groups compared to control group (P<0.001). The F, F+HPsP, and HPsP groups had lower vitamin K-2 and D-3 levels (P<0.001 and P<0.01, respectively). The F, HPsP, F+HPsP, and double-dose groups had higher cholesterol levels (P<0.05, P<0.01, P<0.001, respectively).Conclusion: HPsP extract altered fructose-induced fatty liver composition and levels of reduced glutathione and vitamins A, D, E, and K. Unexpectedly, high liver cholesterol levels require treatment.