Hepatic lipid profiles

Hepatic lipids were extracted according to the method of Folch ²². In brief, extracted buffer A (chloroform : methanol = 2:1, v/v) was added to liver tissues, and the tissues were centrifuged at 3500 ×g and 4 °C for 10 minutes. After removal of the supernatants, the sample was remixed appropriately with extracted buffer B (chloroform : methanol : water = 3:48:47, v/v) and then centrifuged again. Finally, soluble lipids in the subnatant layer was used for further analysis. Hepatic triacylglycerol (TG) and total cholesterol (TC) concentrations were determined using diagnostic kits (Randox Laboratories, Antrim, UK). The liver extraction was mixed well with 100 times reagent buffer for 10 minutes and the absorbance value was measured under OD 500 nm.