Honokiol

Specific pathogen-free female mice (wild type C57BL/6, 3 months old) were purchased from the Guangdong Medical Animal Centre (People's Republic of China). All experiments and protocols were approved by the Ethics Committee of Shantou University Medical College, and all experiments were conducted in accordance with authorities of the Guangdong province. Trimebutine (sc-204928, Santa Cruz) and honokiol (4590, Tocris Bioscience) were dissolved in dimethyl sulfoxide (DMSO; 196055, MP Biomedicals, France) and then diluted in 0.9% saline solution. DMSO needed to be used in the experiments, because the compounds are not soluble in purely physiological vehicles. Trimebutine and honokiol were administered via the tail vein at a dose of 1 mg per kg body weight (total volume 50 μl), respectively. All efforts were made to minimize the suffering of animals and to reduce the number of animals used in the experiments.

3T3-L1 cells were treated with the Sirt3 inhibitor 3-TYP (3-(1H-1,2,3-triazol-4-yl) pyridine) (Selleckchem, Houston, TX, USA) at 50 µM and 100 µM, or with the Sirt3 activator Honokiol (Tocris, Minneapolis, MN 55413, USA) at 1, 5, and 10 µM or were untreated as control and differentiated into adipocytes by the adipocyte differentiation procedure described below. Honokiol and 3-TYP treatments were applied during the process of differentiation of adipocytes (day 1 to day 6).