Regulation of apolipoprotein B production and secretion in response to the change of intracellular cholesteryl ester contents in rabbit hepatocytes.
In the present study, we investigated the mechanisms that regulate apolipoprotein B-100 (apoB) secretion in response to the change of intracellular cholesteryl ester contents by adding low density lipoprotein (LDL) and an inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase (pravastatin) in rabbit hepatocyte culture system. LDL caused a significant dose-dependent increase in apoB secretion. On the other hand, the addition of pravastatin decreased apoB secretion significantly. LDL caused a dose-dependent increase in cellular cholesteryl ester, while cellular cholesteryl ester was decreased by pravastatin significantly. Therefore, these results indicate that the change of apoB secretion was in parallel with the change of cellular cholesteryl ester contents. Cellular contents of free cholesterol, triglyceride, and phospholipid did not change. To investigate intracellular degradation of apoB prior to secretion, pulse-chase experiments were performed. It was shown that the addition of pravastatin accelerated intracellular degradation of apoB, while LDL slowed the apoB intracellular degradation rate. We also investigated whether the change of cellular cholesteryl ester could affect the apoB mRNA level. Northern blot analysis and solution hybridization RNase protection assay demonstrated that neither LDL nor pravastatin caused a significant change in cellular apoB mRNA level. We conclude that intracellular cholesteryl ester contents play a critical role in apoB secretion, and the intracellular apoB degradation rate could be the main mechanism that regulates apoB secretion in response to the change of intracellular cholesteryl ester level.
Department of Geriatric Medicine, Faculty of Medicine, Kyoto University, Japan.
M
M
Tanaka
H
H
Jingami
H
H
Otani
M
M
Cho
Y
Y
Ueda
H
H
Arai
Y
Y
Nagano
T
T
Doi
M
M
Yokode
T
T
Kita
D000085:Acetates / Q000378:metabolism; D000818:Animals; D001055:Apolipoproteins B / Q000096:biosynthesis / Q000378:metabolism*; D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism; D002788:Cholesterol Esters / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000097:blood / Q000302:isolation & purification / Q000494:pharmacology; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D017035:Pravastatin / Q000494:pharmacology; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism; D011817:Rabbits
D000085:Acetates; D001055:Apolipoproteins B; D002250:Carbon Radioisotopes; D002788:Cholesterol Esters; D008077:Lipoproteins, LDL; D012333:RNA, Messenger; D002784:Cholesterol; D017035:Pravastatin