The pool of free cholesterol is not of major importance for regulation of the cholesterol 7 alpha-hydroxylase activity in rat liver microsomes.

The relationship between the cholesterol 7 alpha-hydroxylase activity and the pool of free cholesterol in rat liver microsomes was studied under experimental conditions aimed to stimulate (biliary drainage, cholestyramine treatment, and lymphatic drainage) as well as inhibit (chenodeoxycholic acid treatment) bile acid synthesis. Highly accurate methods based on isotope dilution-mass spectrometry were used both for assay of the cholesterol 7 alpha-hydroxylase activity and the concentration of free cholesterol in the microsomes. In the assay of the cholesterol 7 alpha-hydroxylase, only endogenous cholesterol was used as substrate for the enzyme. Under the experimental conditions employed, the concentration of microsomal free cholesterol remained essentially unchanged in spite of a more than 20-fold variation in enzyme activity. It is concluded that the total pool of free cholesterol in the microsomes is not of major regulatory importance for the cholesterol 7 alpha-hydroxylase in rats.