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Trans-10,Cis-12 conjugated linoleic acid reduces triglyceride content while differentially affecting peroxisome proliferator activated receptor γ2 and aP2 expression in 3T3-L1 preadipocytes

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Lipids

Abstract

A series of experiments was conducted using 3T3-L1 preadipocytes as the cell model to determine: (i) whether the triglyceride (TG)-lowering effects of a crude mixture of conjugated linoleic acid (CLA) isomers were due to a specific isomer of CLA and the timing of treatment, (ii) if CLA reduced TG content by inhibiting a key regulator of adipogenesis, (iii) if CLA incorporated into either neutral lipid or phospholipid cell fractions, and (iv) whether the effects of CLA treatment were reversible. Trans-10,cis-12 CLA reduced TG content, whereas the cis-9,trans-11 isomer increased TG content compared to vehicle [bovine serum albumin (BSA)] controls. Treatment with 50 μM trans-10,cis-12 CLA during the entire 6 d of differentiation reduced TG content to a greater extent than treatment during either the first 3 d or last 3 d of differentiation. Trans-10,cis-12 CLA treatment of preadipocyte cultures for 48 h increased peroxisome proliferator activated receptor γ2 (PPARγ2) protein expression compared to cultures treated with linoleic acid (LA) or the BSA controls. CLA had no effect on adipose P2 (aP2), a fatty acid-binding protein regulated by PPARγ2. Both the cis-9,trans-11 and the trans-10,cis-12 isomers of CLA were incorporated into neutral lipids and phospholipids. However, cis-9,trans-11 CLA levels were one- to twofold higher than trans-10,cis-12 CLA levels. Moreover, trans-10,cis-12 CLA treatment reduced cis-11 18∶1 concentrations in both neutral lipids and phospholipids while increasing cis-9 18∶1 and 18∶2 concentrations. Palmitoleic acid (16∶1) levels were also lower in the neutral lipid fraction of cultures treated with trans-10,cis-12 CLA. Supplementing trans-10,cis-12 CLA-treated cultures (50 μM) with increasing levels of LA resulted in a dose-dependent increase in TG content compared to cultures treated with 50 μM CLA alone. LA supplementation also prevented some of the morphological changes associated with trans-10,cis-12 CLA treatment as seen with scanning electron microscopy. Treatment with 50μM trans-10,cis-12 CLA for 6 d decreased PPARγ2 levels, and supplementation of CLA-treated cultures with LA increased PPARγ2 levels compared with cultures treated with CLA alone. Taken together, these data indicate that in cultures of 3T3-L1 preadipocytes: (i) trans-10,cis-12 CLA is the TG-lowering isomer of CLA, and its effects are dependent on dose, duration of treatment, and the amount of LA in the cultures; (ii) trans-10,cis-12 CLA treatment alters the monounsaturated fatty acid profile of neutral- and phospholipids of the cultures; and (iii) although acute (2−d) trans-10,cis-12 CLA treatment increased PPARγ2 protein levels, chronic (6−d) treatment decreased PPARγ2 levels.

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Abbreviations

AA:

arachidonic acid

ANOVA:

analysis of variance

BCA:

bicinchoninic acid

BSA:

bovine serum albumin

CLA:

conjugated linoleic acid

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

fetal bovine serum

HMDS:

hexamethyldisilazane

IBMX:

isobutylmethylxanthine

LA:

linoleic acid

LPL:

lipoprotein lipase

PBS:

phosphate buffered saline

PPARγ2:

peroxisome proliferator activated receptor γ2

SCD-1:

stearoyl-CoA desaturase-1

SEM:

scanning electron microscopy

TBP:

TFII basal transcription factor

TBS-T:

medium containing Tris HCl+NaCl+Tween 20

TG:

triglyceride

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Author information

Authors and Affiliations

  1. Department of Biology, High Point University, 27410, High Point, North Carolina

    Y. Park & M. Pariza

  2. Food Research Institute, University of Wisconsin at Madison, 53706, Madison, Wisconsin

    L. Curtis & B. Kuebler

  3. Graduate Program in Nutrition, University of North Carolina at Greensboro, P.O. Box 26170, 27402-6170, Greensboro, NC

    M. Evans & M. McIntosh

Authors
  1. M. Evans
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  2. Y. Park
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  3. M. Pariza
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  4. L. Curtis
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  5. B. Kuebler
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  6. M. McIntosh
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Corresponding author

Correspondence to M. McIntosh.

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Evans, M., Park, Y., Pariza, M. et al. Trans-10,Cis-12 conjugated linoleic acid reduces triglyceride content while differentially affecting peroxisome proliferator activated receptor γ2 and aP2 expression in 3T3-L1 preadipocytes. Lipids 36, 1223–1232 (2001). https://doi.org/10.1007/s11745-001-0836-z

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  • DOI: https://doi.org/10.1007/s11745-001-0836-z

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