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Background: White adipose tissue (WAT) browning can promote thermogenesis and could be a promising targetfor treating obesity. Rare ginsenosides transformed from stems and leaves of Panax ginseng (T-GSSL) exhibitnumerous biological activities. However, its potential anti-obesity effects and underlying mechanism remainlargely unknown.

Methods: Five amino acids were selected as the catalysts for the transformation of ginsenosides into rare ginsenosides.

An obese mouse model was established by feeding mice a high-fat diet (HFD) for 14 weeks. The effectsof T-GSSL on obese mice were assessed by measuring body weight, fat mass, energy expenditure (EE), andglucose tolerance. The 3T3-L1 cells were differentiated into mature adipocytes and incubated with T-GSSL.

Immunohistochemistry, co-immunoprecipitation (Co-IP), enzyme-linked immunosorbent assays (ELISA), westernblotting (WB), real-time polymerase chain reaction (PCR), and other methods were used to investigate the targetsand mechanisms of action of T-GSSL.

Results: Ginsenosides in GSSL were hydrolyzed using glutamic acid as a catalyst and 12 rare ginsenosides wereproduced, with a total conversion rate of 95 %. T-GSSL ameliorated metabolic disorders, lipid ectopic deposition,and obesity, and maintained glucose homeostasis in obese mice. T-GSSL treatment promoted adipose browningand enhanced EE in both HFD mice and 3T3-L1 cells. These effects were decreased in cells treated with a proteinkinase A (PKA) antagonist or subjected to PKAcα knockdown, whereas they were increased in REGγ􀀀 /􀀀 mice. Theinhibition of REGγ alongside the activation of the PKA/CREB pathway elucidates the mechanism through whichT-GSSL reverses obesity by promoting the browning of adipose tissue.

Conclusions: T-GSSL attenuates diet-induced obesity by promoting adipose browning through the inhibition ofREGγ and subsequent activation of the PKA/CREB pathway.

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