In vivo regulation of low-density lipoprotein receptor and apolipoprotein B gene expressions by dietary fat and cholesterol in inbred strains of mice

Two proteins that may be important in the hypercholesterolemia and atherosclerosis produced by dietary fat and/or cholesterol are apoB and the LDL-receptor. We evaluated the molecular and genetic regulation of these two proteins by two important components of atherogenic diets: dietary fatty acids and dietary cholesterol. The control diet (C) contained 5% corn oil; the high cholesterol (HC) diets, 5% corn oil plus 0.5% or 2% cholesterol; the high fat diet (HF) 1% corn oil and 20% hydrogenated coconut oil; the fat plus cholesterol diets (HF/C) were the same as HF diet plus either 0.5% or 2% cholesterol. Ten strains of inbred mice were fed the C and HF/C (2% cholesterol) diets. Three strains; C3H, C57BL and SWR, were studied in greater detail. In them the effects of dietary fat and cholesterol were assessed separately and together. These three strains were fed all six diets. Lipoprotein profiles of plasma and indexes of lipoprotein composition were obtained by gel filtration chromatography and in selected strains by gradient ultracentrifugation. Relative rates of transcription of LDL-receptor mRNA and apoB mRNA were measured in purified mouse liver nuclei and levels of LDL-receptor mRNA and apoB mRNA in liver and intestine were quantified by RNA excess solution hybridization assays. The HF/C diet produced rises in plasma total-, VLDL- and LDL-cholesterol and apoB concentrations in the ten strains. VLDL and LDL became cholesterol-enriched and the proportion of total cholesterol transported in VLDL and LDL rose at the expense of HDL. This general pattern of HF/C diet-induced changes was similar in all strains, but there were marked quantitative differences between strains with respect to lipid and lipoprotein concentrations, and compositions and the distribution of cholesterol on both the HC and HF/C diets. The strain-related differences were not due to differences in absorption of dietary cholesterol because, for any given diet, hepatic cholesterol levels increased to the same extent in all strains. Nor were the strain-related differences related to alleles of the apoB gene as determined by RFLP analyses. In the three strains, hepatic LDL-receptor mRNA transcription was suppressed by all diets. But, LDL-receptor mRNA levels in both intestine and liver were suppressed only by the HC and HF/C diets and not by the HF diet. Thus, dietary cholesterol decreased LDL-receptor mRNA levels by mechanisms operating at the transcriptional level, while dietary fatty acids, in addition to inhibiting transcription also appeared to enhance mRNA stability. Dietary cholesterol had no effects or relatively small effects on hepatic apoB mRNA transcription, while the high fat diets (HF and HF/C) significantly increased apoB mRNA transcription. Hepatic apoB mRNA levels showed no consistent changes on any of the diets suggesting that mRNA stability in liver may be affected by dietary fat. By contrast, intestinal apoB mRNA levels increased modestly but significantly on the HF and HF/C diets perhaps reflecting the enhanced transport of fatty acids by the intestine and demonstrating that hepatic and intestinal apoB mRNAs are regulated differently. Thus, the molecular regulation of apoB and LDL-receptor is affected differently by fatty acids and cholesterol and the regulation is tissue specific.