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Non-alcoholic fatty liver disease (NAFLD) is a prevalent chronic liver condition that is strongly linked to gut microbiota imbalance and chronic inflammation. This study aims to explore the preventive effects of dihydromyricetin (DHM) on NAFLD by modulating the intestinal flora and the TLR4/NF-κB signaling pathway. Fifty male C57BL/6J mice were randomly assigned to five groups: a normal control group, a model group, and three DHM treatment groups receiving low (500 mg/kg), medium (750 mg/kg), and high doses (1,000 mg/kg). NAFLD was induced using a high-fat diet, and DHM was administered for 8 weeks. ELISA measured serum levels of LPS, IL-1β, and TNF-α, while Western Blot assessed liver expression of TLR4 and NF-κB p65. Changes in intestinal flora composition were analyzed using high-throughput 16S rRNA sequencing. The results showed that DHM treatment significantly reduced serum levels of LPS, IL-1β, and TNF-α, decreasing the liver expression of TLR4 and NF-κB p65. Intestinal flora analysis indicated a notable increase in beneficial bacteria, especially in the medium and high-dose groups. DHM treatment also significantly improved liver pathology, reducing fat deposition and inflammatory cell infiltration. In conclusion, DHM effectively prevents the progression of NAFLD by improving gut microbiota balance and suppressing inflammatory signaling pathways.

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