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n-3 PUFA as Regulators of Cardiac Gene Transcription: A New Link between PPAR Activation and Fatty Acid Composition

  • Original Article
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Lipids

Abstract

The fatty acids regulate gene expression directly binding to nuclear receptors or affecting the protein content of transcription factors. In this work, supplementing primary cultures of neonatal rat cardiomyocytes with 60 µM EPA or DHA, we demonstrated by an ELISA assay an increased PPAR β/δ binding to DNA. n-3 PUFA supplementation deeply changed the acyl composition of both cytosolic and nuclear fractions. The high content of total fatty acids, particularly EPA and DHA, and its increase following supplementation suggested a selective accumulation of n-3 PUFAs in the nucleus, supporting the direct interaction of n-3 PUFA with PPAR. The activity of acyl-CoA thioesterase (ACOT), catalyzing the reaction leading to NEFA from acyl-CoA, increased in n-3 PUFA supplemented cells. The NEFA/acyl-CoA ratio is an important regulator of the fatty acid transport to the nucleus and consequent modulation of gene transcription, and although ACOT activity is not the only parameter of this ratio, it is important for the control of the NEFA pool composition. Our data further clarify what happens in cardiomyocytes following n-3 PUFA supplementation, linking the modification of acyl composition to ACOT activity and PPAR activation.

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Abbreviations

ACOT:

Acyl-CoA thioesterase

A-FABP:

Adipocyte fatty acid binding protein

B23:

Nucleophosmin

BSA:

Bovine serum albumin

CF:

Cytoplasmic fraction

ChREBP:

Carbohydrate response element-binding protein

CVD:

Cardiovascular diseases

DHA:

Docosahexaenoic acid

DPA:

Docosapentaenoic acid

DTNB:

5,5′-Dithiobis(2-nitrobenzoic acid)

EPA:

Eicosapentaenoic acid

FA:

Fatty acid

FABP:

Fatty acid binding protein

FCS:

Fetal calf serum

HNF-4α:

Hepatocyte nuclear factor 4α

HRP:

Horseradish peroxidase

HS:

Horse serum

K-FABP:

Keratinocyte FABP

LXR:

Liver X receptors

MEK1:

Mitogen-activated protein kinase/extracellular signal-regulated kinase 1

NEFA:

Nonesterified fatty acid

NF:

Nuclear fraction

NF kappa B:

Nuclear factor kappa B

PPAR:

Peroxisome proliferator activated receptor

PPRE:

Peroxisome proliferator responsive element

PUFA:

Polyunsaturated fatty acid

RXR:

Retinoid X receptor

SREBP:

Sterol regulatory element-binding protein

TNB:

2-Nitro-5-thiobenzoic acid

WCL:

Whole cell lysate

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Acknowledgments

This work was partially supported by Granarolo S.p.A. (Bologna, Italy) and by Italian MIUR (RFO 2007). The authors thank Mrs. Fabiana Missiroli for her skillful technical assistance.

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Authors and Affiliations

  1. Department of Biochemistry “G. Moruzzi”, Alma Mater Studiorum, University of Bologna, via Irnerio, 48, 40126, Bologna (BO), Italy

    Mattia Di Nunzio

  2. Food Science Campus, Alma Mater Studiorum, University of Bologna, Piazza Goidanich, 60, 47023, Cesena (FC), Italy

    Francesca Danesi & Alessandra Bordoni

Authors
  1. Mattia Di Nunzio
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  2. Francesca Danesi
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  3. Alessandra Bordoni
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Correspondence to Alessandra Bordoni.

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Di Nunzio, M., Danesi, F. & Bordoni, A. n-3 PUFA as Regulators of Cardiac Gene Transcription: A New Link between PPAR Activation and Fatty Acid Composition. Lipids 44, 1073–1079 (2009). https://doi.org/10.1007/s11745-009-3362-y

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  • DOI: https://doi.org/10.1007/s11745-009-3362-y

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