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Comparative Study
. 2009 Feb;58(2):186-95.
doi: 10.1016/j.metabol.2008.09.012.

Differential influence of different dietary fatty acids on very low-density lipoprotein secretion when delivered to hepatocytes in chylomicron remnants

Iliana López-Soldado  1 , Michael Avella, Kathleen M Botham
Affiliations
Comparative Study

Differential influence of different dietary fatty acids on very low-density lipoprotein secretion when delivered to hepatocytes in chylomicron remnants

Iliana López-Soldado et al. Metabolism. 2009 Feb.

Abstract

The influence of dietary fats carried in chylomicron remnants on the hepatic secretion of very low-density lipoprotein (VLDL) was investigated using chylomicron remnant-like particles (CRLPs) and cultured rat hepatocytes as the experimental model. Chylomicron remnant-like particles containing triacylglycerol (TG) from palm, olive, or corn (enriched in saturated, monounsaturated, or n-6 polyunsaturated fatty acids) oil, respectively, were incubated with cultured hepatocytes for 5 hours. The medium was then removed and replaced with medium without CRLPs; and the secretion of TG, cholesterol, and apolipoprotein B48 during the following 16 hours was determined. Secretion of TG into the d less than 1.050-g/mL fraction containing VLDL was unaffected by olive CRLPs, but was significantly increased in cells exposed to palm or corn CRLPs in comparison with both olive CRLPs and control incubations without CRLPs. Secretion of apolipoprotein B48, however, was not changed by any of the CRLP types. Apolipoprotein B messenger RNA levels were decreased by olive and corn CRLPs, and 3-hydroxy-3-methylglutaryl coenzyme A reductase messenger RNA abundance was increased by palm CRLPs; but expression of other genes involved in the regulation of VLDL secretion was unaffected. These findings demonstrate that CRLPs enriched in saturated fatty acids or n-6 polyunsaturated fatty acids increase the secretion of TG in VLDL, possibly because of the secretion of larger particles, whereas those enriched in monounsaturated fatty acids have no effect. Thus, different dietary fats have differential effects on VLDL secretion directly when delivered to the liver in chylomicron remnants.

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Figures

Fig. 1
Fig. 1
Chylomicron remnant–like particles were prepared as described in “Materials and methods,” and the apo E content was assessed by SDS-PAGE. A, Apolipoprotein E bands in a typical experiment: Lane 1, palm CRLPs; lane 2, olive CRLPs; lane 3, corn CRLPs. B, Quantification by optical density volume analysis. Data shown are the mean ± SEM from 3 separate preparations.
Fig. 2
Fig. 2
Hepatocytes (6 × 106) were incubated with or without CRLPs derived from palm, olive, or corn oil (0.3 μmol TG per milliliter) for 5 hours; the medium was then removed and replaced with fresh medium without CRLPs; and the incubation was continued for a further 16 hours. After this time, the medium was collected; and the content of TG in the medium or in the d less than 1.050–g/mL fraction was determined. Data are expressed as nanomoles per milligram cell protein and are the mean ± SEM from 8 separate hepatocyte preparations, each performed in duplicate. Significant limits: *P < .05 vs control (incubation without CRLPs); †P < .05 vs corn CRLPs; #P < .05 vs palm CRLPs.
Fig. 3
Fig. 3
Hepatocytes (6 × 106) were incubated with or without CRLPs derived from palm, olive, or corn oil (0.3 μmol TG per milliliter) for 5 hours; the medium was then removed and replaced with fresh medium without CRLPs; and the incubation was continued for a further 16 hours. After this time, the cells were harvested; and their content of TG, TC, UC, and CE was determined. Data are expressed as nanomoles per milligram cell protein and are the mean ± SEM from 9 separate hepatocyte preparations, each performed in duplicate. Significant limits: *P < .05 vs control (incubation without CRLPs); †P < .05 vs corn CRLPs.
Fig. 4
Fig. 4
Hepatocytes (6 × 106) were incubated with or without CRLPs derived from palm, olive, or corn oil (0.3 μmol TG per milliliter) for 5 hours; the medium was then removed and replaced with fresh medium without CRLPs; and the incubation was continued for a further 16 hours. After this time, the medium was collected; and the content of apo B48 was determined. A, Apolipoprotein B48 bands in a typical experiment. B, Quantification by Western blotting coupled to ECL. Data are expressed as the percentage of the control in arbitrary units per milligram cell protein and are the mean ± SEM from 8 separate hepatocyte preparations, each performed in duplicate. Lane 1, positive control containing apo B48 and apo B100; lane 2, palm CRLPs; lane 3, olive CRLPs; lane 4, corn CRLPs; lane 5, without CRLPs.
Fig. 5
Fig. 5
Hepatocytes (1.2 × 106) were incubated with or without CRLPs derived from palm, olive, or corn oil (0.3 μmol TG per milliliter) for 5 or 16 hours. The medium was then removed, the total RNA was extracted from the cells, and the relative abundance of transcripts for (A) apo B and (B) HMG-CoA reductase was determined by qPCR. Values are expressed as fold change relative to the control (hepatocytes incubated without CRLPs). Data are the mean ± SEM from 8 separate hepatocyte preparations, each performed in duplicate. *P < .05 vs control (2-way analysis of variance).
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