Discharge of newly‐synthesized dolichol and ubiquinone with lipoproteins to rat liver perfusate and to the bile

Abstract

An effective system for perfusing rat liver using complete tissue culture medium and washed calf erythrocytes as oxygen carriers was devised. Infusion of taurocholate and glucose proved necessary to maintain stable metabolic activity and bile secretion during a 6-hr period. Perfusate pO2, pCO2 and pH values were monitored continuously and found to be stable. Electron microscopic examination revealed the maintenance of normal hepatic structure, even after 6 hr. Normal rates of protein and urea synthesis, no leakage of cytoplasmic enzymes, and continuous bile acid production demonstrated the functional integrity of this system. Using [3H]mevalonic acid as precursor, dolichol, dolichyl phosphate, ubiquinone and cholesterol were found to be continuously synthesized in this perfused liver system. All these lipids appeared in the perfusate, indicating discharge through the ER-Golgi system. The lipoproteins of the perfusate were isolated by ultracentrifugation and characterized with respect to size distribution and lipid composition. Dolichol was found in VLDL, LDL and HDL fractions, with the highest concentration present in the latter. In rat and human blood plasma this lipid was mainly associated with HDL. The ubiquinone in the perfusate was primarily associated with the VLDL fraction, while in rat plasma it was found more evenly distributed among all the three lipoprotein fractions studied. Dolichol, ubiquinone and cholesterol were also discharged to the bile, whereas dolichyl phosphate was not. Thus, newly-synthesized dolichol and ubiquinone are transported out of the hepatocyte to the blood and to the bile.