The increasing prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) and the limited treatment options highlight the need for effective alternatives. Natural bioactive compounds are increasingly valued for their safety and multifunctional regulatory effects. 10-hydroxy-2(E)-decenoic acid (10-HDA), a unique fatty acid from royal jelly, exhibits diverse biological activities and potential benefits for liver health. In this study, we investigated the hepatoprotective effects of 10-HDA and its underlying mechanisms using palmitic acid/oleic acid-treated HepG2 cells and high-fat diet-induced golden hamsters. 10-HDA significantly reduced hepatic triglyceride and cholesterol contents, improved serum lipid profiles, and alleviated steatosis. Integrated transcriptomic and metabolomic analysis revealed 10-HDA regulates lipid homeostasis mainly through activating AMPK signaling and regulating cholesterol metabolism, including downregulation of   HMGCR, SORT1, and SOAT1, and upregulation of ABCG5/8 and CYP7A1. AMPKα knockdown abolished these effects, confirming its key role in mediating the metabolic benefits of 10-HDA.  These findings demonstrate that10-HDA protects against MASLD by modulating hepatic cholesterol metabolism via AMPK activation, highlighting its potential as a dietary bioactive compound for metabolic health.

Fatty acids are distinctive components of royal jelly (RJ), serving as a crucial indicator for assessing the quality of RJ. This study aimed to establish a rapid and simultaneous quantification method for various fatty acids in RJ utilizing gas chromatography. An optimized two-step extraction process, incorporating ethanol and diethyl ether followed by derivatization with N,O-bis-(trimethylsilyl) trifluoroacetamide, was developed to enhance sensitivity and precision in detecting fatty acids. Validation of the methodology revealed excellent linearity (R² > 0.999), precision (relative standard deviation (RSD) < 1%), repeatability (RSD < 1%), and recoveries (94.4%–104%). Furthermore, the limits of detection and quantification were found to be low. The results indicated that the method offered reliable and consistent quantification of major fatty acids, thereby improving quality control for RJ and facilitating its applications in food, pharmaceutical, and biochemical research.