Lutein and Zeaxanthin Provide Protection Against Aflatoxin B1-Induced Liver and Kidney Toxicity: Insights from in Silico and in Vivo Studies

Exposure to aflatoxin B ₁ (AFB ₁ ) poses significant threats to food safety, increases food insecurity, and endangers public health due to its pronounced organ toxicity and carcinogenicity, especially affecting the hepatorenal system. This study integrates network pharmacology and molecular docking analyses to investigate the protective roles of the natural carotenoids lutein (LUT) and zeaxanthin (ZEA) against AFB ₁ -induced toxicity. Key hub genes-ALB, IL2, LGALS3, SRC, REN, EGFR, PRKACA, LCN2, F2, and TTR- implicated in carcinogenesis, stress response, cellular homeostasis, and tissue remodelling were identified via Network Pharmacology analysis. Molecular docking demonstrated that LUT (-9.0 kcal/mol) and ZEA (-9.5 kcal/mol) have higher affinity to inflammation-related proteins (LGALS3) than AFB1 (-8.4 kcal/mol), suggesting their strong anti-fibrotic and antioxidant potentials. Lutein also showed the highest binding affinity for REN (-9.5 kcal/mol) compared to AFB ₁ (-8.8 kcal/mol), suggesting a specific mechanism through which LUT may influence the RAAS pathway to reduce AFB ₁ -induced kidney damage. In vivo experiments using Wistar rats (N = 25; 10 weeks old, weighing 220 ± 20 g) randomly assigned to five groups and treated as follows: Control group (corn oil only, 2 mL/kg), AFB1 only (75 µg/kg), LUT/ZEA only (100 mg/kg), AFB1 (75 µg/kg) plus LUT/ZEA1 (100 mg/kg), and LUT/ZEA2 (200 mg/kg) per os for 28 days. And followed by the assessments of liver and kidney function, serum lipid profiles, enzymatic and non-enzymatic antioxidant levels, oxidative stress markers, inflammatory mediators, DNA damage, and apoptosis biomarkers were conducted using spectrophotometric methods, confirming that LUT/ZEA administration alleviated AFB ₁ -induced liver and kidney damage, reduced oxidative stress, inflammation, DNA damage, and apoptosis in a dose-dependent manner. Histopathological analyses further validated these protective effects. Overall, the study highlights the potential of LUT and ZEA as natural agents for mitigating AFB ₁ toxicity, offering promising strategies to improve food safety and reduce health risks associated with mycotoxin exposure.