Early works by Augustinsson (1-4) demonstrated that three types of esterases exist in vertebrate plasma in regard to substrate specificity and susceptibility to selective inhi bitors, and he classified them into arylesterases [EC 3. 1. (5) suggested that decrease in the activity of phenyl acetate esterase (corre sponding to arylesterases, according to the Augustinsson's classification) was correlated with the degree of liver damage and that, clinically, the activity of the enzyme in serum decreased greatly in the case of liver cirrhosis. In the present paper, changes in the total activity and the electrophoretic pattern of arylesterases were studied in CC14-induced mouse liver injury using fl-naphthyl acetate as substrate.
METHODSExperiment I: Effect of CCl4 administration upon electrophoretic pattern of arylesterases in liver 12,000 X g supernatant and serum Male inbred mice of the NC strain weighing 20 to 24 g were divided into 7 groups of 5 animals each, and treated with 1 ml/kg of CC14 in trap eritoneally except the control group. They were sacrificed at 1/4, 3, 6, 24, 48 and 72 hours after CCI4 injection by ex sanguination from the femoral artery. The liver was immediately removed, rinsed with ice-cold physiological saline and homogenized with an equal amount of 0.05 M phosphate buffer, pH 7.4, by a glass homogenizer with a teflon pestle under refrigeration . The subsequent procedures were performed at fC. The homogenate was centrifuged for 45 minutes at 12,000 X g and the supernatant was separated. The blood collected was also centrifuged for 15 minutes at 3,000 r.p.m. to obtain serum.Thin-layer agar gel electrophoresis described by Ogita (6-8) was performed for 2 hours with the constant current adjusted to 1 mA/cm to separate arylesterases in the super natant of liver homogenate and serum. After electrophoresis, the glass plate covered with agar gel film was immersed into phosphate buffer, pH 6.8, containing Q-naphthyl acetate as substrate and naphthanyl diazo blue B and kept at 37°C for 20 minutes. The