Regulation of lipid metabolism-related gene expression in whole blood cells of normo- and dyslipidemic men after fish oil supplementation

Simone Schmidt, Janina Willers, Frank Stahl, Kai-Oliver Mutz, Thomas Scheper, Andreas Hahn, Jan Philipp Schuchardt, Simone Schmidt, Janina Willers, Frank Stahl, Kai-Oliver Mutz, Thomas Scheper, Andreas Hahn, Jan Philipp Schuchardt

Abstract

Background: Beneficial effects of omega-3 polyunsaturated fatty acids (n-3 PUFAs) on the lipid levels of dyslipidemic subjects are widely described in the literature. However, the underlying molecular mechanisms are largely unknown. The aim of this study was to investigate the effects of n-3 PUFAs on the expression of lipid metabolism-related genes in normo- and dyslipidemic men to unveil potential genes and pathways affecting lipid metabolism.

Methods: Ten normo- and ten dyslipidemic men were supplemented for twelve weeks with six fish oil capsules per day, providing 1.14 g docosahexaenoic acid and 1.56 g eicosapentaenoic acid. The gene expression levels were determined by whole genome microarray analysis and quantitative real-time polymerase chain reaction.

Results: Several transcription factors (peroxisome proliferator-activated receptor α (PPARα), retinoid X receptor (RXR) α, RXRγ, hepatic nuclear factor (HNF) 6, and HNF1ß) as well as other genes related to triacylglycerol (TG) synthesis or high-density lipoprotein (HDL-C) and cholesterol metabolism (phospholipids transfer protein, ATP-binding cassette sub-family G member 5, 2-acylglycerol O-acyltransferase (MOGAT) 3, MOGAT2, diacylglycerol O-acyltransferase 1, sterol O-acyltransferase 1, apolipoprotein CII, and low-density lipoprotein receptor) were regulated after n-3 PUFA supplementation, especially in dyslipidemic men.

Conclusion: Gene expression analyses revealed several possible molecular pathways by which n-3 PUFAs lower the TG level and increase the HDL-C and low-density lipoprotein level, whereupon the regulation of PPARα appear to play a central role.

Trial registration: ClinicalTrials.gov (ID: NCT01089231).

Figures

Figure 1
Figure 1
Protein levels of apolipoprotein B48 in normolipidemic and dyslipidemic men. Serum protein levels of apolipoprotein B48 (Apo B48) was determined by ELISA in normo- and dyslipidemic men before (t0) and after twelve weeks (t12) of fish oil supplementation. Differences between t0 and t12 protein levels were tested by a paired t-test, and differences between groups at each time point were tested by unpaired t-test using the statistical package R version 2.15.0.
Figure 2
Figure 2
Regulatory effects of eicosapentaenoic acid and docosahexaenoic acid on lipid metabolism related genes. The Figure presented is based on the analysis of gene expression changes after fish oil supplementation in dyslipidemic male subjects. While bold text and black and grey arrows present findings discovered in this thesis, grey arrows symbolize that until now it has not been clarified if genes are target genes of PPARα. In addition, broken arrows symbolize further possible mechanisms of action based on findings from the literature.
Figure 3
Figure 3
Transcript levels of target genes in normolipidemic and dyslipidemic men. Transcript levels of apolipoprotein CII (Apo CII) and low-density lipoprotein receptor (LDLR) was determined by qRT-PCR in normo- and dyslipidemic men before (t0) and after twelve weeks (t12) of fish oil supplementation. Pooled group samples were used in triplicates. Triplicates were averaged and corrected by two reference genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ribosomal proteine S2 (RPS2). Corrected expressions were compared with baseline gene expression of normolipidemic subjects and relative expression changes are displayed.

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