Oral doses of α-retinyl ester track chylomicron uptake and distribution of vitamin A in a male piglet model for newborn infants.

α-Retinol has utility in determining chylomicron trafficking of vitamin A to tissues given that it will not be recirculated in blood on retinol binding protein (RBP). In this study, α-retinol was used as a chylomicron tag to investigate short-term uptake from high-dose supplements given to piglets as a model for neonates. The distribution of orally administered α-retinol doses in liver and extrahepatic tissues was assessed at varying times after dosing. Male piglets (n = 24 per group) from vitamin A-depleted sows were orally given 26.2 or 52.4 µmol of α-retinyl acetate, the molar equivalent of 25,000 and 50,000 IU of vitamin A, respectively. Tissues were collected and analyzed by HPLC. Lung (6.46 ± 2.94 nmol/g), spleen (22.1 ± 11.3 nmol/g), and adrenal gland (17.0 ± 11.2 nmol/g) α-retinol concentrations peaked at 7 h after dosing, and, by 7 d, α-retinol was essentially cleared from these tissues (≤0.25 ± 0.12 nmol/g). This demonstrates that the lung, spleen, and adrenal gland receive substantial vitamin A from chylomicra to maintain concentrations. Conversely, storage of α-retinol in the liver reached a plateau at 24 h (1.72 ± 0.58 µmol/liver) and was retained through 7 d (2.10 ± 0.38 µmol/liver) (P > 0.05). This indicates that α-retinol was not substantially utilized locally in the liver nor transported out from the liver via RBP. In serum, the majority of α-retinol was in the ester form, which confirms that α-retinol does not bind to RBP but does circulate. α-Retinyl esters were detectable at 7 d in the serum but were not different from baseline. Collectively, these data suggest that crucial immune organs need constant dietary intake to maintain vitamin A concentrations because α-retinol was quickly taken up by tissues and decreased to baseline in all tissues except long-term storage in the liver.

The effects of dietary n-3 polyunsaturated fatty acids delivered in chylomicron remnants on the transcription of genes regulating synthesis and secretion of very-low-density lipoprotein by the liver: modulation by cellular oxidative state.

The influence of chylomicron remnants enriched in n-3 or n-6 polyunsaturated fatty acids (PUFA) (derived from fish or corn oil, respectively) on the expression of mRNA for four genes involved in the regulation of the synthesis, assembly, and secretion of very-low-density lipoprotein (VLDL) in the liver was investigated in normal rat hepatocytes and after manipulation of the cellular oxidative state by incubation with N-acetyl cysteine (NAC) or CuSO(4). The four genes investigated were those encoding apolipoprotein B (apoB), the microsomal triacylglycerol transfer protein (MTP), and the enzymes acyl coenzyme A:diacylglycerol acyltransferase (DGAT) and acyl coenzyme A:cholesterol acyltransferase 2 (ACAT2), which play a role in the regulation of triacylglycerol and cholesteryl ester synthesis, respectively. mRNA levels for apoB, MTP, and DGAT were unaffected by either fish or corn oil chylomicron remnants, but the amount of ACAT2 mRNA was significantly reduced after incubation of the hepatocytes with fish oil remnants as compared with corn oil remnants or without remnants. These findings indicate that the delivery of dietary n-3 PUFA to hepatocytes in chylomicron remnants downregulates the expression of mRNA for ACAT2, and this may play a role in their inhibition of VLDL secretion. However, when the cells were shifted into a pro-oxidizing or pro-reducing state by pretreatment with CuSO(4) (1 mM) or NAC (5 mM) for 24 hr, levels of mRNA for MTP were increased by about 2- or 4-fold, respectively, by fish oil remnants, whereas corn oil remnants had no significant effect. Fish oil remnants also caused a smaller increase in apoB mRNA in comparison with corn oil remnants in NAC-treated cells (+38%). These changes would be expected to lead to increased VLDL secretion rather than the decrease associated with dietary n-3 PUFA in normal conditions. These findings suggest that relatively minor changes in cellular redox levels can have a major influence on important liver functions such as VLDL synthesis and secretion.

Comparison of the uptake and processing of cholesterol from chylomicrons of different fatty acid composition in rats fed high-fat and low-fat diets.

The fate of [3H]cholesterol carried in chylomicrons prepared from rats given a meal of palm oil (rich in long-chain saturated fatty acids), olive oil (rich in monounsaturated fatty acids) or corn oil (rich in n-6 polyunsaturated fatty acids) was investigated in vivo in rats fed a low-fat diet or a diet supplemented with the corresponding oil (to provide 40% of the calories) for 21 days. In the low-fat-fed groups, radioactivity was removed from the blood and secreted into bile over 180 min more rapidly when the chylomicrons were derived from corn oil as compared to palm or olive oil. After feeding the corresponding high-fat diets, however, both parameters were decreased in rats fed palm and corn oil, but not olive oil. As a result of these changes, the rates of removal of radioactivity from the blood and secretion into bile were similar in animals given the olive oil and corn oil diets, and higher than those in rats fed the palm oil diet. All the high-fat diets tended to increase the proportion of the radioactivity in the plasma found in the 1.006-1.050-g/ml fraction (low-density lipoprotein) and decrease that in the 1.050-1.25-g/ml (high-density lipoprotein) fraction in comparison to the respective low-fat diet groups, but the transfer of radioactivity to the plasma high-density lipoprotein fraction was particularly slow in palm-oil-fed rats. These findings indicate that diets high in saturated or n-6 polyunsaturated fat retard the metabolism of chylomicron cholesterol in comparison to diets low in fat, while those high in monounsaturated fat do not have this effect. As a consequence of this, the rate of removal of cholesterol of dietary origin from the body is slower in animals fed saturated as compared to monounsaturated or n-6 polyunsaturated fat. Thus, differential metabolism of chylomicron cholesterol clearly plays an important role in the hyper- and hypo-cholesterolaemic effects of these dietary fats.