Abstract / Summary
Metabolic dysfunction-associated steatotic liver disease (MASLD) often coexists with cardiac dysfunction, calling for therapies that protect both organs. Anethole trithione (ADT), a clinical choleretic agent and sustained-release H₂S donor, has shown hepato-cardiac protective potential, but its efficacy and underlying mechanisms against MASLD and related cardiac impairment remain poorly defined. This study evaluated the therapeutic effects of ADT using high-fat diet (HFD)-induced hamster models with early and advanced MASLD. Syrian golden hamsters were fed an HFD for 6 or 18 weeks to induce early- and advanced-stage MASLD, with daily ADT gavage. Indicators related to liver injury, cardiac function, lipid deposition, and oxidative stress were assessed using biochemical assays, GC-MS, Oil Red O staining, H&E staining, Masson’s trichrome staining, Sirius red staining, immunofluorescence, and echocardiography. Mechanistic studies included quantitative proteomics, qRT-PCR, immunoblotting and gain-of-function experiments. In hamsters with short-term HFD-induced MASLD, ADT markedly reduced serum and hepatic lipid accumulation, decreased liver injury markers, and improved hepatic histology. In long-term HFD-fed hamsters, ADT further alleviated hepatic steatosis, mitochondrial dysfunction, ROS accumulation, and liver fibrosis while preserving cardiac function. Proteomic analysis identified RTN3 as a key mediator, whose upregulation by HFD was reversed by ADT. RTN3 overexpression in THLE-2 hepatocytes and primary cardiomyocytes blunted the lipid-lowering effects of ADT. Mechanistically, ADT inhibited the hepatic SREBP1/ACC1/FASN/SCD1 lipogenic axis and restored IDH2-dependent mitochondrial homeostasis, both downstream of RTN3. In cardiomyocytes, ADT attenuated fatty acid incorporation into lipid droplets in an RTN3-dependent pathway. These findings indicate that ADT relieves MASLD and cardiac dysfunction via RTN3‑dependent regulation of lipid synthesis and oxidative catabolism.