Obesity is characterized by hypertrophy and/or by the differentiation or adipogenesis of pre-existing adipocytes. In this study, we investigated the inhibitory effects of theobromine, a type of alkaloid in cocoa, on adipocyte differentiation of 3T3-L1 preadipocytes and its mechanisms of action. Theobromine inhibited the accumulation of lipid droplets, the expression of PPARγ and C/EBPα, and the mRNA expression of aP2 and leptin. The inhibition of adipogenic differentiation by theobromine occurred primarily in the early stages of differentiation. In addition, theobromine arrested the cell cycle at the G0/G1 phase and regulated the expressions of CDK2, p27, and p21. Theobromine treatment increased AMPK phosphorylation and knockdown of AMPKα1/α2 prevented the ability of theobromine to inhibit PPARγ expression in the differentiating 3T3-L1 cells. Theobromine reduced the phosphorylation of ERK and JNK. Moreover, the secretion and the mRNA level of TNF-α and IL-6 were inhibited by theobromine treatment. These data suggest that theobromine inhibits adipocyte differentiation during the early stages of adipogenesis by regulating the expression of PPARγ and C/EBPα through the AMPK and ERK/JNK signaling pathways in 3T3-L1 preadipocytes.
Four functional constituents, oxyresveratrol 3′-O-β-D-glucoside (ORTG), oxyresveratrol (ORT), t-resveratrol (RT), and moracin (MC) were isolated from the ethanolic extract of mulberry (Morus alba L.) twigs by a series of isolation procedures, including solvent fractionation, and silica-gel, ODS-A, and Sephadex LH-20 column chromatographies. Their chemical structures were identified by NMR and FABMS spectral analysis. Quantitative changes of four phytochemicals in mulberry twigs were determined by HPLC according to cultivar, producing area, and heat processing. ORTG was a major abundant compound in the mulberry twigs, and its levels ranged from 23.7 to 105.5 mg% in six different mulberry cultivars. Three other compounds were present in trace amounts (<1 mg/100 g) or were not detected. Among mulberry cultivars examined, “Yongcheon” showed the highest level of ORTG, whereas “Somok” had the least ORTG content. Levels of four phytochemicals in the mulberry twigs harvested in early September were higher than those harvested in early July. Levels of ORTG and ORT in the “Cheongil” mulberry twigs produced in the Uljin area were higher than those produced in other areas. Generally, levels of ORTG and ORT in mulberry twigs decreased with heat processing, such as steaming, and microwaving except roasting, whereas those of RT and MC did not considerably vary according to heat processing. These results suggest that the roasted mulberry twigs may be useful as potential sources of functional ingredients and foods.
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