Long-chain polyunsaturated fatty acids upregulate LDL receptor protein expression in fibroblasts and HepG2 cells.

The objective of this study was to investigate the effect of individual PUFAs on LDL receptor (LDLr) expression in human fibroblasts and HepG2 cells, and to evaluate whether acyl CoA:cholesterol acyltransferase (ACAT) and sterol regulatory element-binding protein 1 (SREBP-1) were involved in the regulation of LDLr expression by fatty acids. When fibroblasts and HepG2 cells were cultured with serum-free defined medium for 48 h, there was a 3- to 5-fold (P < 0.05) increase in LDLr protein and mRNA levels. Incubation of fibroblasts and HepG2 cells in serum-free medium supplemented with 25-hydroxycholesterol (25OH-cholesterol, 5 mg/L) for 24 h decreased LDLr protein and mRNA levels by 50-90% (P < 0.05). Arachidonic acid [AA, 20:4(n-6)], EPA [20:5(n-3)], and DHA [22:6(n-3)] antagonized the depression of LDLr gene expression by 25OH-cholesterol and increased LDLr protein abundance 1- to 3-fold (P < 0.05), but had no significant effects on LDLr mRNA levels. Oleic (18:1), linoleic (18:2), and alpha-linolenic acids [18:3(n-3)] did not significantly affect LDLr expression. ACAT inhibitor (58-035, 1 mg/L) attenuated the regulatory effect of AA on LDLr protein abundance by approximately 40% (P < 0.05), but did not modify the regulatory effects of other unsaturated fatty acids in HepG2 cells. The present results suggest that AA, EPA, and DHA increase LDLr protein levels, and that ACAT plays a role in modulating the effects of AA on LDLr protein levels. Furthermore, the effects of the fatty acids appeared to be independent of any change in SREBP-1 protein.

Conjugated linoleic acid upregulates LDL receptor gene expression in HepG2 cells.

Conjugated linoleic acid (CLA) exerts anticarcinogenic and antiatherosclerotic effects in animals. The present study was conducted to examine the effects of CLA on LDL receptor (LDLr) expression in HepG2 cells, and to evaluate whether the sterol response element binding protein 1 (SREBP-1) and acyl CoA:cholesterol acyltransferase (ACAT) were involved in the regulation of LDLr expression by CLA. When HepG2 cells were cultured with serum-free DMEM for 48 h, there was a three- to fivefold (P<0.05) increase in LDLr protein and mRNA levels. Incubation of HepG2 cells in serum-free medium supplemented with 25-hydroxycholesterol (25OH, 5 mg/L) for 24 h decreased LDLr protein and mRNA by 50-70% (P<0.05) and mature SREBP-1 by 20-40% (P<0.05). CLA, but not linoleic acid, antagonized the depressive effects of 25OH and increased both LDLr protein and mRNA abundance twofold (P<0.05). LDLr protein and mRNA abundance were not different when HepG2 cells were cultured with CLA (0.4 mmol/L) plus 25OH in the presence or absence of an ACAT inhibitor (58-035, 1 mg/L). Furthermore, CLA had no effect on SREBP-1 abundance. These results suggest that CLA upregulates LDLr expression via a mechanism that is independent of ACAT and SREBP-1.

Iron status in association with cardiovascular disease risk in 3 controlled feeding studies.

BACKGROUND: The role of body iron stores in free radical-induced peroxidation and cardiovascular disease risk has been debated, but controlled feeding studies using measurements of non-transferrin-bound iron (NTBI) and LDL oxidation have not been conducted. OBJECTIVE: We tested the hypothesis that NTBI and other measures of iron status do not affect oxidative susceptibility in healthy subjects with normal iron status. DESIGN: Plasma samples were analyzed from 77 healthy men and women aged 20-65 y who participated in 3 controlled feeding studies in which the type and amount of dietary fat were controlled. Iron status and in vitro LDL oxidation were assessed at baseline and at the end of each feeding period (4-8 wk). RESULTS: No significant relations were found between any measure of iron status (ferritin: 83 +/- 8.9 micro g/L; iron: 20.9 +/- 5.4 micro mol/L; TIBC: 74.4 +/- 11.0 micro mol/L; NTBI: 0.184 +/- 0.15 micro mol/L) and the in vitro measures of LDL oxidation (total dienes: 485 +/- 55 micro mol/mg LDL protein; lag time: 51.7 +/- 15.9 min; and rate of oxidation: 25.4 +/- 6.8 micro mol dienes.min(-1).g LDL protein(-1)). Equal-iron peanut butter-based diets were associated with higher plasma iron in men (22.4 +/- 3.8 micro mol/L) than was the olive oil diet (17.7 +/- 4.5 micro mol/L) (P = 0.02), but this slight elevation did not alter LDL oxidation. CONCLUSIONS: Diet composition may affect plasma iron in men, but LDL oxidative susceptibility is unaffected by the subtle variation in iron status. Thus, the results do not support a relation between iron status and LDL oxidative susceptibility, a possible risk factor for cardiovascular disease.