Rats were treated with ST-638 (30 or 100 mg/kg)

Rats hepatocytes were isolated by in vitro collagenase perfusion and low speed centrifugation. Cell viability was determined by trypan blue exclusion and ranged from 85 to 95%. The hepatocytes were cultured for 24 h in monolayers in a plastic dish (1x10^5 cells/cm2) in Williams’ medium E containing 10% fetal calf serum, 10 nM insulin, 10 nM dexamethasone and 5 KIU/ml aprotinin under 5%CO2 atmosphere. After removal of the medium by aspiration, monolayers of hepatocytes in the dish were further incubated for 0—90 min in Williams’ medium E containing 2% bovine serum albumin with or without the addition of heparin in the presence of ST-638 (1-5mM). The hepatocytes were harvested and centrifuged at 50xg for 5 min to remove cellular debris. The obtained supernatant served as the preparation for the released HTGL activity. HTGL activity was determined by a method using glycerol tri[1-14C]oleate (1.2mkM; 2.5 kBq/ml) as a substrate