Deoxyglucose uptake study

Uptake glucose activity was analyzed by measuring 2-deoxyglucose (2DG) uptake by 3T3-L1 cells according to the manufacturer’s instruction. A mature adipocyte 3T3-L1 cells were incubated for 1 hours with (+)-catechin 10.75; 21.5; 43 µM, proanthocyanidin fraction 25, 50, & 100 µg/mL, and 1 mM metformin in PBS. The luminescence signal was detected as a relative light unit (RLU) on the Promega luminometer SA3030 (Madison, Wiscosin, USA).

In order to investigate the inhibitory activity of 2DG uptake through sirtuin-1 on adipocytes, mature adipocytes 3T3-L1 were treated with sirtuin-1 specific inhibitors, nicotinamide. The nicotinamide 30 mM was used in this assay based on preliminary study.Nicotinamide was added 30 min before addition of (+)-catechin, proanthocyanidin fraction, and metformin.¹³ Calculation of 2DG6P concentration was based on the 2DG6P standard curve.