Terpenoid-enriched Curcuma wenyujin nanovesicles for suppressing inflammation and restoring lipid homeostasis in MASH

Aim: This study investigates the potential of Curcuma wenyujin-derived nanovesicles (CW-DNVs) to ameliorate metabolic dysfunction-associated steatohepatitis (MASH) and explores their underlying mechanism, focusing on hepatic macrophage accumulation and the regulation of lipid metabolism. Methods: CW-DNVs were isolated via ultracentrifugation and sucrose gradient purification, and their physicochemical properties, cellular uptake, and in vivo biodistribution were characterized. Anti-inflammatory and lipid-lowering effects were evaluated in liver macrophages, hepatocytes, and a high-fat diet (HFD)-induced MASH mouse model. Lipidomic, small-molecule, and small RNA (sRNA) cargoes were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and RNA sequencing. Results: CW-DNVs were spherical (~211 nm diameter), had a zeta potential of -27.4 mV, and were enriched in lipids, proteins, sRNAs, and terpenoids like well-known bioactive curcumenol and germacrone. Following intraperitoneal injection, they preferentially accumulated in Kupffer cells and were cleared within 7 days. In HFD-fed mice, CW-DNVs reduced body weight gain, hepatic steatosis, serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglycerides (TG), total cholesterol (TC), and hepatic interleukin (IL)-6 levels. Mechanistically, they upregulated peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and microsomal triglyceride transfer protein (MTTP), and downregulated fatty acid synthase (FASN), promoting lipid oxidation and export. In vitro, CW-DNVs suppressed lipopolysaccharide-activated IL-6, IL-1β, and tumor necrosis factor-alpha (TNF-α) in macrophages and reduced oleic acid-induced lipid accumulation in hepatocytes. sRNA sequencing identified predominantly rRNA-derived fragments (not canonical miRNAs); however, anti-inflammatory activity was primarily attributed to terpenoid components. Conclusion: CW-DNVs exert dual functionality in modulating macrophage inflammation and lipid metabolism. Loaded with bioactive terpenoids, they represent an effective natural nanoplatform for MASH therapy.