Nelumbinis Plumula attenuates high-fat-induced inflammation in non-alcoholic fatty liver through the NOX/NF-κB pathway

ETHNOPHARMACOLOGICAL RELEVANCE: Non-alcoholic fatty liver disease (NAFLD) is a prevalent chronic hepatic metabolic disorder and represents a major public health concern worldwide. Nelumbinis Plumula, a traditional Chinese herbal medicine, has attracted significant interest given its high concentration of alkaloids and flavonoids, known to modulate lipid metabolism (LM) and exert antioxidant and anti-inflammatory effects. Although its extract ENP has been reported to govern LM, the mechanism remains unclear. AIM OF THE STUDY: To investigate whether ENP could ameliorate hepatic inflammation and free fatty acid (FFA)-induced lipid deposition in NAFLD models and to explore the mechanisms underlying its therapeutic effects. METHODS: High-performance liquid chromatography (HPLC) was used to identify the major constituents of ENP. NAFLD was modeled in vivo by feeding mice a high-fat diet (HFD) for 20 weeks and in vitro by exposing AML12 cells to FFA. ENP was administered at various doses to evaluate its pharmacodynamic effects. Network pharmacology combined with metabolomics was applied to identify potential targets and signaling pathways, and these findings were further validated in the above NAFLD models to assess ENP's efficacy in regulating LM. RESULTS: Four alkaloids and eight flavonoids were identified as the primary active compounds of ENP. In mice, ENP administration significantly attenuated HFD-induced increases in body weight, epididymal fat, liver hypertrophy, and histopathological damage, while improving LM and liver function. Integrated network pharmacology and metabolomics analyses revealed that the modulation of oxidative stress and inflammatory pathways, particularly through the NF-κB signaling cascade, was central to ENP's therapeutic effects. Consistently, ex vivo experiments revealed that ENP restored lipid metabolic balance in NAFLD models by suppressing inflammation via the NOX/NF-κB signaling pathway. CONCLUSION: ENP attenuates NAFLD-associated inflammatory responses by inhibiting NOX/NF-κB signaling, providing a theoretical foundation for the therapeutic application of Nelumbinis Plumula.