Comparative analysis of genistein and treadmill exercise interventions on high-fat high-sucrose diet-induced oxidative stress and vascular dysfunction in male mice

Background: High fat, high sucrose diets (H) affect metabolic health and oxidative stress (OS). OS can lead to many diet-related diseases and tissue damage. Genistein, a primary isoflavone, has positive effects on lipid metabolism and oxidative stress. It is shown to improve antioxidant capacity and reduce hepatic fat accumulation, obesity, diabetes mellitus, and hypercholesterolemia. Aerobic exercise can also activate the body’s antioxidant system and reduce OS, depending on mode, intensity, and duration of exercise. TBARS (Thiobarbituric Acid Reactive Substances) is an indicator of lipid peroxidation and OS. H diets decrease antioxidants and increase OS markers like TBARS, resulting in OS. Hypothesis: We hypothesized that the combination of genistein and treadmill exercise would better prevent vascular damage and OS compared to either intervention alone. Method: Adult male mice were fed standard chow (Lean) or a high fat high sucrose diet (H). Animals fed H were further divided into groups treated with or without genistein (G) (600 mg/kg diet) and/or treadmill exercise (X) consisting of 30-minute running sessions five days per week. We then utilized immunohistochemistry and tissue staining to analyze differences in lipids (Oil Red O staining), morphology (H&E), and protein expression of uncoupling protein 1 (UCP1), endothelial and inducible nitric oxide synthase (eNOS and iNOS, respectively) in the aorta. TBARS were measured in serum samples collected from each animal. Results: Oil Red O staining showed that lean mice had minimal lipid accumulation, H groups showed increased lipids, whereas H+G and H+X interventions prevented lipid accumulation. iNOS protein expression was increased in H compared to lean controls and the intervention groups, showing that G and/or X prevent H-induced increases in iNOS. Lean mice show normal patterns of H&E staining, intervention groups show improved tissue organization compared to H mice. UCP1 expression was increased in H mice compared to lean controls, while intervention groups show different expression patterns of this thermogenic protein. eNOS protein expression was increased in H compared to lean mice, while intervention groups had intermediate levels of expression. Serum TBARS was comparable across groups (Kruskal-Wallis, P=0.422). Two-way ANOVA for the four H groups revealed significant effects of genistein, with lower TBARS in H mice treated with G compared to untreated H mice. Exercise had no significant effect (P=0.205) on TBARS and there were no interactions between genistein treatment and exercise (P=0.237). Conclusion: While limited by small sample sizes, our findings indicate that genistein supplementation prevents H-induced OS and vascular disorder whereas exercise had no effect on OS but did protect the vasculature. Funding was provided by Midwestern University (LA). This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.