Mechanisms of Mulberry Leaf Extracts Ameliorating Inflammation and Fibrosis in Non‐Alcoholic Fatty Liver Disease Mice via <scp>NF‐κB</scp> and <scp>TGFβ1/Smad3</scp> Pathways

Mulberry leaf, as a traditional Chinese medicinal plant, has been utilized in the treatment of various diseases, including diabetes, cardiovascular diseases, inflammatory disorders, and liver diseases. However, the mechanisms underlying its therapeutic effects on non-alcoholic fatty liver disease (NAFLD) remain unclear. Therefore, this study aims to investigate the potential mechanisms of mulberry leaf extract (MLE) in the treatment of NAFLD. The chemical composition of MLE was analyzed using ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The NAFLD mice model was induced by a high-fat, high-fructose, and high-cholesterol (HFFC) diet, followed by intervention with MLE. The results indicated that the administration of MLE notably reduced the obesity (p < 0.05), oxidative stress (p < 0.05), inflammation (p < 0.05), and ECM deposition (p < 0.05) induced by the HFFC diet, restored the parameters of liver function, and attenuated the pathological changes. Utilizing a combination of integrated liver non-targeted metabolomics, network pharmacology, and transcriptomic approaches, we deciphered the molecular mechanisms by which MLE exerted its therapeutic effects in the treatment of NAFLD. In detail, our findings revealed that MLE suppressed the TGFβ1/Smad3 and NF-κB signaling pathways to ameliorate fibrosis and inflammation. This study provided novel insights into the correlation between MLE and NAFLD progression, offering a scientific foundation for the prospective use of MLE in the treatment of NAFLD.