In vitro stimulation of rat liver retinyl ester hydrolase by ethanol.

Retinyl ester hydrolase (REH), the enzyme which converts retinyl esters to retinol, was partially characterized from whole liver homogenates of rats using an HPLC method with quantitation of retinol product. Optimal results were obtained by incubation of 1 mg of whole homogenate protein with 900 microM all-trans-retinyl palmitate and 275 mM 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate in a 0.1 M Tris-maleate buffer, pH 7.0, for 1 h at 37 degrees C. The enzyme assay proved to be sensitive and reproducible, with an interanimal coefficient of variation of 13% (n = 7). Because ethanol has been shown to mobilize vitamin A from the liver, we tested its effect on REH activity at several concentrations. In concentrations ranging from 0.01 to 0.5 M, ethanol added in vitro caused a concentration related increase in REH activity (from 20 to 86% above baseline activity). This increase was specific to ethanol as acetaldehyde, 1-propanol, and t-butanol either did not change or significantly decreased REH activity over the range of concentrations tested. The range of concentrations of ethanol causing stimulation in our assays was within the range of concentrations seen in the blood of rats after acute ethanol ingestion. Stimulation of REH activity could explain, in part, the well-known effects of ethanol on mobilization of vitamin A from liver stores.