Dual Mechanistic Anti-Obesity Effects of Red Okra and <i>Diospyros lotus</i> Mixture via Fat Binding and AMPK-Mediated Lipid Metabolism

Obesity is a major global health issue associated with metabolic dysfunctions including dyslipidemia, insulin resistance, and hepatic steatosis. This study investigated the dual anti-obesity mechanisms of a combined extract of red okra (<i>Abelmoschus esculentus</i> L. Moench, RO) and <i>Diospyros lotus</i> L. leaves (DL) in a high-fat diet (HFD)-induced obese mouse model. Fat-binding capacity (FBC) analysis revealed that RO exhibited significantly higher FBC than DL, suggesting a greater potential to inhibit intestinal fat absorption. <i>In vitro</i>, RO more effectively suppressed lipid accumulation in 3T3-L1 adipocytes, whereas DL enhanced lipolytic activity by stimulating glycerol release from differentiated adipocytes. To optimize the complementary functions of both extracts, various mixing ratios were evaluated, and the 4:1 ratio (RODL^TM) was identified as optimal. This combination resulted in approximately 25% reduction in lipid accumulation, a significant increase in glycerol release, and a 39% elevation in fecal fat excretion. In HFD-fed mice, oral administration of the RODL mixture led to up to 14% suppression in body weight gain, improved serum triglyceride and cholesterol levels, and alleviated hepatic steatosis. Mechanistically, the extract combination activated AMPKα signaling, leading to the upregulation of lipolysis-related proteins (ATGL, pHSL), thermogenesis-related markers (UCP1, PGC1α), and fatty acid oxidation regulators (PPARα, CPT1, ACOX1). These findings indicate that the RO and DL extract combination exerts anti-obesity effects through both inhibition of fat absorption and modulation of lipid metabolic pathways. This dual mechanism supports its potential as a safe and effective natural therapeutic for obesity prevention and management.