Esterification of cholesterol in high density lipoprotein decreases its ability to support ACTH-stimulated steroidogenesis by rat adrenocortical cells.
Addition of high density lipoprotein 3 (HDL3) isolated from human plasma of d greater than 1.125 g/ml which had been preincubated for 24 h at 37 degrees C enhanced steroidogenesis by cultured rat adrenal cells only 38% as well as HDL3 isolated from unincubated plasma. Loss of steroidogenic activity due to preincubation was associated with a decrease in the percent HDL3 cholesterol remaining unesterified. Inhibition of lecithin-cholesterol acyltransferase activity by heating (60 degrees C, 1 h) or addition of dithionitrobenzoic acid (1.4 mM) prevented esterification of cholesterol in HDL and also prevented loss of steroidogenic activity. Although incubation of plasma of d greater than 1.125 g/ml prior to isolation caused cholesterol esterification, there was no change in the ratio of total cholesterol to protein in HDL, size and shape of the HDL particle as assayed by measurement of sedimentation velocity, nor affinity for the putative HDL receptor. Addition of unesterified cholesterol to preincubated HDL restored steroidogenic activity. These results indicate that unesterified cholesterol in HDL is preferentially used as substrate for rat adrenal steroidogenesis. The effects of nonlipoprotein serum proteins on HDL action in the adrenal were also examined. The ability of HDL3 to enhance rat adrenal steroidogenesis was not significantly less in serum-free media than in media supplemented with lipoprotein-poor fetal calf serum or human plasma of d greater than 1.21 g/ml, suggesting that rat adrenal uptake of HDL cholesterol does not depend on participation of plasma enzymes or transport proteins.
Gwynne
J T
JT
Department of Medicine, University of North Carolina, Chapel Hill 27514.
Mahaffee
D D
DD
D000302:Adrenal Cortex / Q000187:drug effects / Q000378:metabolism*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters / Q000378:metabolism*; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D005260:Female; D006023:Glycoproteins; D006801:Humans; D008075:Lipoproteins, HDL / Q000378:metabolism*; D053440:Lipoproteins, HDL3; D051381:Rats; D011919:Rats, Inbred Strains; D013256:Steroids / Q000096:biosynthesis
C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters; D006023:Glycoproteins; D008075:Lipoproteins, HDL; D053440:Lipoproteins, HDL3; D013256:Steroids; D000324:Adrenocorticotropic Hormone; D002784:Cholesterol; D004228:Dithionitrobenzoic Acid