Reduced cholesterol esterification in CaCo-2 cells by indirect action of pravastatin

• Published in: Atherosclerosis Vol. 125; no. 1; pp. 63 - 70
• Main Authors: Reimann, Frank M., Winkelmann, Friederike, Fellermann, Klaus, Stange, Eduard F.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Ireland Ltd 23.08.1996; Elsevier
• Subjects: Acyl-CoA cholesteryl acyltransferase; Biological and medical sciences; CaCo-2 cell; Caco-2 Cells; Cell Division - drug effects; Cell Survival - drug effects; Cholesterol; Cholesterol - metabolism; Esterification - drug effects; General and cellular metabolism. Vitamins; HMG-CoA reductase inhibitor; Humans; Intestinal Mucosa - metabolism; Intestines - drug effects; LDL; Lipoproteins - metabolism; Liposomes - metabolism; Medical sciences; Mevalonic Acid - analogs & derivatives; Mevalonic Acid - pharmacology; Microsomes - metabolism; Pharmacology. Drug treatments; Pravastatin; Pravastatin - pharmacology; Sterol O-Acyltransferase - metabolism; Tetrazolium salts; Acyl-CoA cholesteryl acyltransferase; LDL; CaCo-2 cell; Tetrazolium salts; HMG-CoA reductase inhibitor; Pravastatin; Cholesterol; Cell culture; Decrease; Enzyme; Enzyme inhibitor; Hydroxymethylglutaryl-CoA reductase; Oxidoreductases; Biological activity; Esterification; Antilipemic agent
• ISSN: 0021-9150; 1879-1484
• DOI: 10.1016/0021-9150(96)05848-0

In microsomal preparations of CaCo-2 cells pravastatin decreased cholesterol esterifying activity at 25 μg/ml to 82.5% and at 800 μg/ml to 56.2% of controls. Pravastatin reduced cholesteryl ester formation dose-dependently also in viable CaCo-2 cells. However, the maximal inhibition was by 90.4% at pravastatin concentration of 25 μg/ml, half maximal inhibition occurred between concentrations of 5 and 10 μg/ml. Addition of mevalonolactone, which serves as endogenous source of cholesterol, antagonized this effect. At 10 mM mevalonolactone (MVL) even doses up to 200 μg/ml of pravastatin were ineffective. On the other hand, pravastatin suppressed cholesteryl ester formation when acyl-CoA cholesterol acyltransferase (ACAT) (E.C. 2.3.1.26) activity was stimulated by addition of exogenous liposomal or Low Density Lipoprotein (LDL)-derived cholesterol. This inhibition was refractory to increasing amounts of exogenous cholesterol up to 400 μg/ml. Therefore we conclude that only excessive doses of pravastatin suppress ACAT activity directly. In viable cells the observed inhibition of cholesteryl ester formation is due to the block in de novo synthesis of cholesterol, causing a lack of substrate for ACAT and of non-sterol products of mevalonic acid. Furthermore pravastatin interferes with the esterification and/or intracellular transport only of exogenous cholesterol, confirming former results of a compartmentalized cholesterol metabolism in the enterocyte.