Integrated Network Pharmacology and Gut Microbiota Analysis Reveals the Alcoholic Extract of Anacyclus pyrethrum Root Prevents Nonalcoholic Fatty Liver Disease via the LPS/TLR4/NF-κB Pathway.

The global incidence of nonalcoholic fatty liver disease (NAFLD) is rising, with no approved pharmacotherapy available. Medicinal plants offer a potential preventive strategy. Anacyclus pyrethrum root exhibits anti-inflammatory and glucose-regulating properties, but its role in NAFLD prevention is unclear. This study aims to investigate the preventive effect of Anacyclus pyrethrum root ethanol extract (APE) against NAFLD and its underlying mechanisms. The chemical composition of APE was analyzed by UHPLC-HRMS. Network pharmacology predicted the potential signaling pathways underlying its protective effects against NAFLD. In a 12-week high-fat diet mice model, APE treatment led to measurements of blood glucose, lipid profiles, liver function parameters, histopathological changes in liver and colon, and gut microbiota alterations via 16S rDNA sequencing. In animal experiments, APE lowered fasting and random blood glucose, total cholesterol, triglycerides, LDL-C, AST, ALT, and serum lipopolysaccharide while increasing HDL-C, and alleviated hepatic steatosis. Network pharmacology suggested APE acts via TLR, NF-κB, and TNF pathways. In vivo, APE suppressed hepatic TLR4, MyD88, p-NF-κB p65, the p-NF-κB p65/NF-κB p65 ratio, and TNF-α/IL-6 levels. Gut microbiota analysis showed increased Akkermansiaceae and decreased Desulfovibrionaceae. APE also upregulated intestinal Occludin and ZO-1, and downregulated intestinal TNF-α and IL-6. APE prevents NAFLD progression, potentially by regulating gut microbiota, protecting the intestinal mucosal barrier, and inhibiting the LPS/TLR4/MyD88/NF-κB pathway.