Heyehui attenuates diet-induced obesity via regulating LEP/AMPK/ACC axis and restoring intestinal function in mice

Obesity, a global epidemic characterized by excessive adipose accumulation and disrupted metabolic homeostasis, is closely linked to type 2 diabetes and non-alcoholic fatty liver disease. Lotus leaf (Nelumbo nucifera Gaertn.) is a traditional edible and medicinal plant, and its calcined derivative, Heyehui (Hyh), has been documented for anti-obesity effects since the Ming Dynasty. However, the chemical composition and molecular mechanisms of Hyh against diet-induced obesity (DIO) remain largely uncharacterized.

This study aimed to identify the chemical constituents and quantify the key bioactive components of Hyh, validate its anti-obesity efficacy in a DIO mouse model, and elucidate the underlying molecular mechanisms, thereby providing modern scientific evidence for its traditional application in anti-obesity therapy.

Hyh composition was profiled by UPLC-MS, UV-Vis, and HPLC. DIO mice received Hyh intervention for 8 weeks; body weight, obesity indices, and serum glucose-lipid parameters were then assessed alongside histopathological examinations. The LEP/AMPK/ACC pathway and lipid synthesis factors were quantified by ELISA, immunofluorescence, qRT-PCR, and Western blot. Active components targeting LEPR were screened via molecular docking and DeepDrugDiscovery. Nuciferine and quercetin were further evaluated in 3T3-L1 adipocytes with shLEPR knockdown and Compound C validation. Intestinal barrier integrity and gut microbiota were analyzed by 16S rRNA sequencing. Fecal SCFAs were quantified by GC.

Hyh was rich in alkaloids and flavonoids, with high nuciferine and quercetin contents. Hyh-treated DIO mice exhibited significantly reduced body weight, Lee's index, abdominal circumference, and body fat, accompanied by improved serum glucose-lipid profiles. Histologically, adipocyte shrinkage, decreased hepatic lipid deposition, and downregulated lipid synthesis factors were observed in adipose and liver tissues. Hyh activated the LEP/AMPK/ACC axis, promoting fatty acid β-oxidation; nuciferine and quercetin were identified as the key mediators, as evidenced by shLEPR knockdown and Compound C reversal. Hyh also upregulated occludin, claudin1, and ZO-1 in the duodenum, restored intestinal barrier integrity, suppressed CD36 and FATP4, increased gut microbial diversity, elevated fecal SCFA levels, and enriched genera such as Rikenella, which negatively correlated with obesity indicators.

Hyh, with nuciferine and quercetin as its principal active constituents, ameliorates obesity and metabolic disorders in DIO mice through a multi-target mechanism that encompasses suppression of lipid synthesis, activation of fatty acid β-oxidation, repair of the intestinal barrier, remodeling of the gut microbiota, and elevation of short-chain fatty acid levels. The present study characterized the chemical composition of Hyh and elucidated its anti-obesity mechanisms, thereby providing a modern scientific basis for its traditional application. Although its clinical efficacy remains to be validated in future trials, Hyh demonstrates considerable potential as a natural anti-obesity agent.