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Research Article|Open Access
Flavonoid-Rich mulberry leaf extract modulate lipid metabolism, antioxidant capacity, and gut microbiota in high-fat diet-induced obesity: potential roles of FGF21 and SOCS2
Guangxi Key Laboratory of Environmental Exposure Omics and Life Cycle Health, College of Public Health, Guilin Medical University, Guilin 541199, China
Guangxi Key Laboratory of Drug Discovery and Optimization, College of Pharmacy, Guilin Medical University, Guilin 541199, China
Guangxi Eco-engineering Vocational and Technical College, School of Tourism and Traffic Management, Liuzhou 545004, China
Liuzhou Institute of Technology, Liuzhou Special Food Flavor and Quality Control Research Center of Engineering Technology, Liuzhou 545616, China
†These authors contributed equally to this work.
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Highlights
(1) Network pharmacology analysis identified key targets such as IL6, STAT3, FXR, PPAR, and CYP family, revealing the potential mechanisms of mulberry leaves in NAFLD treatment.
(2) Mulberry leaf aqueous extract (MLQE) intervention significantly reduced body weight, improved lipid profiles, and alleviated hepatic steatosis in high-fat diet-induced obese mice, with notable histological and metabolic improvements.
(3) MLQE enhanced hepatic and plasma antioxidant activities, elevating catalase (CAT), glutathione peroxidase (GSH-Px), and HDL-C levels, while reducing oxidative stress markers.
(4) RNA-seq and Western Blot analyses demonstrated that MLQE normalized metabolic gene expressions, upregulating FXR and SHP, and correcting FGF21 and SOCS2 downregulation, with positive modulation of gut microbiota composition.
Graphical Abstract
Mulberry leaves are a rich source of active ingredients exhibiting notable antioxidant activity and have been demonstrated to possess hypolipidemic and hypoglycemic effects. Non-alcoholic fatty liver disease (NAFLD), the most prevalent chronic liver disease, is increasingly prevalent worldwide, with a strong correlation between NAFLD occurrence and obesity. We aimed to investigate the association between the active ingredient targets in mulberry leaves and stems and the disease targets of NAFLD/NASH through network pharmacology, and designed animal experiments to verify our findings. In addition to the normal and model groups, three intervention groups were established for the experiment: the mulberry leaf aqueous extract (MLQE), Orlistat, and swimming exercise groups. The intervention effect of MLQE on NAFLD was observed by comparing it with widely recognized weight loss drugs and exercise interventions. The results demonstrated that all three interventions effectively reduced body weight and body fat, exhibited robust antioxidant capacity, and significantly improved the dysbiosis of gut microbiota induced by a high-fat diet. Histological analysis also supported the conclusions of lipid-lowering, anti-inflammatory, and amelioration of liver injury. Through RNA-seq analysis, we identified the key genes FGF21 and SOCS2 that may be associated with the effective intervention of mulberry leaves in NAFLD, and validated the genes and proteins related to NAFLD and lipid metabolism by qPCR and WB analysis. Our study found that MLQE has an anti-NAFLD effect not inferior to orlistat and swimming exercise, indicating that it is a promising candidate for the treatment of NAFLD. Meanwhile, FGF21 and SOCS2 may be key targets for the treatment of NAFLD, and more studies are needed to verify this.
Abstract
Non-alcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease in clinical practice. This study examined the effects of mulberry leaf aqueous extract (MLQE) on lipid metabolism and gut microbiota in mice with high-fat diet-induced NAFLD. Using network pharmacology, we identified key targets of mulberry leaf and stem’s constituents for NAFLD regulation and validated their ameliorative effects through animal experiments. Our results demonstrated that interventions with orlistat (HFO), swimming exercise (HFS), and MLQE (HFM) significantly reduced weight gain, lipid accumulation, and oxidative stress in mace compared to the high-fat control (HFC) group. Histopathological analysis revealed improvements in liver steatosis, hepatocyte arrangement, inflammation, and adipocyte deformation. Molecular analyses indicated decreased mRNA levels of lipid synthesis genes (sterol regulatory element-binding protein 1 (SREBP1), bile salt export pump (BSEP), small heterodimer partner (SHP), and stearoyl-CoA desaturase-1 (SCD1)) and increased protein levels of farnesoid X receptor (FXR) and SHP in the intervention groups, consistent with RNA-seq findings. Additionally, RNA-seq analysis showed upregulation of fibroblast growth factor 21 (FGF21) and suppressor of cytokine signaling 2 (SOCS2), aligning with protein level observations. The interventions also modulated gut microbiota composition, reducing the Firmicutes-to-Bacteroidetes (F/B) ratio and adjusting the abundance of Acetatifactor, Roseburia, and Parasutterella. These findings suggest MLQE as a promising therapeutic approach for NAFLD, with lipid-lowering, antioxidative, and gut microbiota-regulating effects. FGF21 and SOCS2 may be key targets in NAFLD treatment.
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