Mulberry Leaf Extracts Ameliorate Aflatoxin B 1 ‐Induced Liver Injury by Modulating Nrf2 Pathway and Gut Microbiota

ABSTRACT Mulberry leaf, an edible medicinal plant, is traditionally recognized for its diverse biological functions, including antioxidant and anti‐inflammatory effects. Aflatoxin B 1 (AFB 1 ), a prevalent foodborne mycotoxin, causes oxidative stress, inflammation, and liver injury. However, the role of functional components of mulberry leaf in AFB 1 ‐induced liver injury is still elusive. This study investigated the hepatoprotective potential of mulberry complex extract (MCE) and mulberry leaf polysaccharide (MLP) in a murine model of AFB1‐induced liver injury. Twenty‐four male mice were randomly divided into four groups, exposed to AFB 1 (0.75 mg/kg), and treated with MCE (500 mg/kg) or MLP (300 mg/kg) for four weeks. Both treatments significantly improved liver function and reduced hepatic oxidative stress and inflammation. Additionally, they markedly inhibited NLRP3/Caspase‐1/GSDMD‐mediated hepatocyte pyroptosis. These benefits were associated with activation of the Nrf2 pathway, upregulation of antioxidant genes and phase II detoxification enzymes, restoration of intestinal barrier integrity, and modulation of gut microbiota to enhance short‐chain fatty acid production. Notably, MCE and MLP interventions increased Nrf2 gene expression by 2.8‐ and 2.3‐fold, respectively. This study provides new evidence supporting the hepatoprotective effects of mulberry leaf extracts against AFB 1 ‐induced toxicity. The findings highlight the potential of MCE and MLP as promising natural therapeutic agents for preventing or mitigating mycotoxin‐related liver damage through modulation of oxidative stress, inflammation, pyroptosis, and gut‐liver axis interactions. Practical Applications Based on this research, mulberry leaf extracts could be developed into dietary supplements or functional food ingredients to help protect the liver in people regularly exposed to aflatoxin‐contaminated foods. Additionally, they show promise as natural additives in animal feed to reduce liver damage and improve the health of livestock consuming contaminated feedstuffs, potentially leading to safer animal products.