Title: Differential inhibition of hepatic, pancreatic, and plasma fatty acid ethyl ester synthase by tri-o-tolylphosphate in rats.

Authors: Mericle, Kelly A; Kaphalia, Bhupendra S; Ansari, G A S

Published In Toxicol Appl Pharmacol, (2002 Mar 1)

Abstract: Fatty acid conjugation of alcohols, catalyzed by fatty acid ethyl ester synthase (FAEES), results in the formation of lipophilic esters. Although the activity of FAEES is reported in almost all organs, including plasma, the interrelationship among various proteins expressing FAEES activity in different organs/tissues is not well understood. Earlier, we have reported an inhibition of FAEES activity in human hepatoma cells by tri-o-tolylphosphate (TOTP; serine esterase inhibitor). The present study was undertaken to further characterize the hepatic, plasma, and pancreatic FAEES in rats after ip injection of 10, 25, 50, or 100 mg/kg TOTP in corn oil or vehicle alone. After 18 h, animals were euthanized and FAEES activity in the plasma and postnuclear fractions of hepatic and pancreatic homogenates were assayed by measuring the ester formation following incubation with [1-(14)C]oleic acid and ethanol or methanol as substrates. Significant inhibition of FAEES activity was observed in hepatic postnuclear fraction. The esterase activity also showed a pattern similar to fatty acid ethyl and methyl ester synthesizing activity. A trend similar to hepatic synthesizing and hydrolyzing activities was also found in the plasma of TOTP-treated rats. However, no inhibition of synthetic activity toward formation of fatty acid ethyl or methyl esters or p-nitrophenyl acetate hydrolyzing activity was observed in the pancreas of rats after TOTP exposure. Our results also show that the protein expressing FAEES activity in the pancreas does not cross-react with antibodies to rat adipose tissue FAEES using Western blot analysis, which recognizes approximately 60- and approximately 84-kDa proteins in the liver and plasma, respectively. Furthermore, the inhibition in liver is at the functional level of enzyme as no change was observed between control and treated animals by immunohistochemistry. We conclude that fatty acid ethyl or methyl ester synthesizing enzyme(s) in the liver and plasma, which are inhibited by TOTP, are different from that present in the pancreas.

PubMed ID: 11884245

MeSH Terms: Acyltransferases/antagonists & inhibitors*; Acyltransferases/blood; Animals; Blotting, Western; Comparative Study; Electrophoresis, Polyacrylamide Gel; Immunohistochemistry; Liver/drug effects; Liver/enzymology*; Male; Organ Specificity; Pancreas/drug effects; Pancreas/enzymology*; Rats; Rats, Sprague-Dawley; Research Support, U.S. Gov't, P.H.S.; Tritolyl Phosphates/toxicity*