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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

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

References

  1. Belury, M. (1995) Conjugated Dienoic Linoleate: A Polyunsaturated Fatty Acid with Unique Chemoprotective Properties, Nutr. Rev. 53, 83–89.

    PubMed  CAS  Article  Google Scholar 

  2. Lee, K., Kritchevsky, D., and Pariza, M. (1994) Conjugated Linoleic Acid and Atherosclerosis in Rabbits, Atherosclerosis 108, 19–25.

    PubMed  CAS  Article  Google Scholar 

  3. Nicolosi, R., Rogers, E., Kritchevsky, D., Scimeca, J., and Huth, P. (1997) Dietary Conjugated Linoleic Acid Reduces Plasma Lipoproteins and Early Aortic Atherosclerosis in Hypercholesterolemic Hamsters, Artery 22, 266–277.

    PubMed  CAS  Google Scholar 

  4. Houseknecht, K., Heuvel, J., Moya-Camarena, S., Portocarrero, C., Peck, L., Nickel, K., and Belury, M. (1998) Dietary Conjugated Linoleic Acid Normalizes Impaired Glucose Tolerance in Zucker Diabetic Fatty fa/fa Rat, Biochem. Biophys. Res. Commun. 244, 678–682.

    PubMed  CAS  Article  Google Scholar 

  5. West, D., Delany, J., Camet, P., Blohm, F., Truett, A., and Scimeca, J. (1998) Effects of Conjugated Linoleic Acid on Body Fat and Energy Metabolism in the Mouse, Am. J. Physiol. 275, R667-R672.

    PubMed  CAS  Google Scholar 

  6. Park, Y., Albright, K., Liu, W., Storkson, J., Cook, M., and Pariza, M. (1997) Effect of Conjugated Linoleic Acid on Body Composition in Mice, Lipids 32, 853–858.

    PubMed  CAS  Google Scholar 

  7. Park, Y., Albright, K., Storkson, J., Liu, W., Cook, M., and Pariza, M. (1999) Changes in Body Composition in Mice During Feeding and Withdrawal of Conjugated Linoleic Acid, Lipids 33, 243–248.

    Google Scholar 

  8. Park, Y., Storkson, J., Albright, K., Liu, W., and Pariza, M. (1999) Evidence That Trans-10,cis-12 Isomer of Conjugated Linoleic Acid Induces Body Composition Changes in Mice, Lipids 34, 235–41.

    PubMed  CAS  Google Scholar 

  9. Tsuboyama-Kasaoka, N., Takahashi, M., Tanemura, K., Kim, H., Tange, T., Okuyama, H., Kasai, M., Ikemoto, S., and Ezaki, O. (2000) Conjugated Linoleic Acid Supplementation Reduces Adipose Tissue by Apoptosis and Develops Lipodystrophy in Mice, Diabetes 49, 1534–1542.

    PubMed  CAS  Google Scholar 

  10. DeLany, J., Blohm, F., Truett, A., Scimeca, J., and West, D. (1999) Conjugated Linoleic Acid Rapidly Reduces Body Fat Content in Mice Without Affecting Energy Intake, Am. J. Physiol. 276, R1172-R1179.

    PubMed  CAS  Google Scholar 

  11. Cook, M., Jerome, D., Crenshaw, T., Buege, P., Pariza, M., Al-bright, K., Schmidt, S., Scimeca, J., Lotgren, P., and Hentges, E. (1999) Feeding Conjugated Linoleic Acid Improves Feed Efficiency and Reduces Carcass Fat in Pigs, in 27th Steenbock Symposium: Adipocyte Biology and Hormone Signaling Symposium, p. 26, Department of Biochemistry, University of Wisconsin, Madison.

    Google Scholar 

  12. Ostrowska, E., Muralitharan, M., Cross, R., Bauman, D., and Dunshea, F. (1999) Dietary Conjugated Linoleic Acids Increase Lean Tissue and Decrease Fat Deposition in Growing Pigs, J. Nutr. 129, 2037–2042.

    PubMed  CAS  Google Scholar 

  13. Gavino, V., Gavino, G., Leblanc, M., and Tuchweber, B. (2000) An Isomeric Mixture of Conjugated Linoleic Acid but Not Pure cis-9,trans-11 Octadecadienoic Acid Affects Body Weight Gain and Plasma Lipids in Hamsters, J. Nutr. 130, 27–29.

    PubMed  CAS  Google Scholar 

  14. West, D., Blohm, F., Truett, A., and Delany, J. (2000) Conjugated Linoleic Acid Persistently Increases Total Energy Expenditure in AKR/J Mice Without Increasing Uncoupling Protein Gene Expression, J. Nutr. 130, 2471–2477.

    PubMed  CAS  Google Scholar 

  15. Blankson, H., Stakkestad, J., Fagertun, H., Thom, E., Wadstein, J., and Gudmundsen, O. (2000) Conjugated Linoleic Acid Reduces Body Fat Mass in Overweight and Obese Humans, J. Nutr. 130, 2943–2948.

    PubMed  CAS  Google Scholar 

  16. Zambell, K., Keim, N., Van Loan, M., Gale, B., Benito, P., Kelley, D., and Nelson, G. (2000) Conjugated Linoleic Acid Supplementation in Humans: Effects on Body Composition and Energy Expenditure, Lipids 35, 777–782.

    PubMed  CAS  Google Scholar 

  17. Satory, D., and Smith, S. (1999) Conjugated Linoleic Acid Inhibits Proliferation but Stimulates Lipid Filling of Murine 3T3-L1 Preadipocytes, J. Nutr. 129, 92–97.

    PubMed  CAS  Google Scholar 

  18. Brodie, A., Manning, V., Ferguson, K., Jewell, D., and Hu, C.Y. (1999) Conjugated Linoleic Acid Inhibits Differentiation of Pre-and postconfluent 3T3-L1 Preadipocytes but Inhibits Cell Proliferation Only in Pre-Confluent Cells, J. Nutr. 129, 602–606.

    PubMed  CAS  Google Scholar 

  19. Evans, M., Geigerman, C., Cook, J., Curtis, L., Kuebler, B., and McIntosh, M. (2000) Conjugated Linoleic Acid Suppresses Triglyceride Content and Induces Apoptosis in 3T3-L1 Preadipocytes, Lipids 35, 899–910.

    PubMed  CAS  Google Scholar 

  20. Choi, Y., Kim, Y., Han, Y., Park, Y., Pariza, M., and Ntambi, J. (2000) The trans-10,cis-12 Isomer of Conjugated Linoleic Acid Downregulates Stearoyl-CoA Desaturase 1 Gene Expression in 3T3-L1 Adipocytes, J. Nutr. 130, 1920–1924.

    PubMed  CAS  Google Scholar 

  21. MacDougald, O., and Lane, D. (1995) Transcriptional Regulation of Gene Expression During Adipocyte Differentiation, Annu. Rev. Biochem. 64, 345–373.

    PubMed  CAS  Article  Google Scholar 

  22. McIntosh, M., Lea-Currie, R., Geigerman, C., and Pasevouros, L. (1999) Dehydroepiandrosterone Alters the Growth of Stromal-Vascular Cells from Human Adipose Tissue, Int. J. Obesity 23, 595–602.

    CAS  Article  Google Scholar 

  23. Cornelius, P., MacDougald, O., and Lane, D. (1999) Regulation of Adipocyte Differentiation, Annu. Rev. Nutr. 14, 99–129.

    Article  Google Scholar 

  24. Folch, J., Lees, M., and Sloane Stanley, G. (1957) A Simple Method for the Isolation and Purification of Total Lipides from Animal Tissues, J. Biol. Chem. 226, 497–509.

    PubMed  CAS  Google Scholar 

  25. Juaneda, P., and Rocquelin, G. (1985) Rapid and Convenient Separation of Phospholipids and Nonphosphorus Lipids from Rat Heart Using Silica Cartridges, Lipids 20, 40–41.

    PubMed  CAS  Google Scholar 

  26. Baumgard, L., Corl, B., Dwyer, D., Saebo, A., and Bauman, D. (2000) Identification of the Conjugated Linoleic Acid Isomer That Inhibits Milk Fat Synthesis, Am. J. Physiol. 278, R179-R184.

    CAS  Google Scholar 

  27. Casimir, D., and Ntambi, J. (1996) Preadipocyte Differentiation Blocked by Prostaglandin Stimulation of FP Receptor, Differentiation 60, 203–210.

    PubMed  CAS  Article  Google Scholar 

  28. Aihaud, G., Grimaldi, P., and Negrel, R. (1992) Cellular and Molecular Aspects of Adipose Tissue Development, Annu. Rev. Nutr. 12, 207–233.

    Article  Google Scholar 

  29. Morrison, R., and Farmer, S. (2000) Hormonal Signaling and Transcriptional Control of Adipocyte Differentiation, J. Nutr. 130, 31165–31215.

    Google Scholar 

  30. Jones, S., Ma, D., Robinson, F., Field, C., and Clandinin, M. (2000) Isomers of Conjugated Linoleic Acid (CLA) Are Incorporated into Egg Yolk Lipids by CLA-Fed Laying Hens, J. Nutr. 130, 2002–2005.

    PubMed  CAS  Google Scholar 

  31. Szymczyk, B., Pisulewski, P., Szczurek, W., and Hanczakowski, P. (2000) The Effects of Feeding Conjugated Linoleic Acid (CLA) on Rat Growth Performance, Serum Lipoproteins and Subsequent Lipid Composition of Selected Rat Tissues, J. Sci. Food Agric. 80, 1553–1558.

    CAS  Article  Google Scholar 

  32. Bretillon, L., Chardigny, J., Gregoire, S., Berdeaux, O., and Sébédio, J. (1999) Effects of Conjugated Linoleic Acid Isomers on the Hepatic Microsomal Desaturation Activities in vitro, Lipids 34, 965–969.

    PubMed  CAS  Google Scholar 

  33. Ip, M., Masso-Welch, P., Shoemaker, S., Shea, W., and Ip, C. (1999) Conjugated Linoleic Acid Inhibits Proliferation and Induces Apoptosis of Normal Rat Mammary Epithelial Cells in Primary Culture, Exp. Cell Res. 250, 22–34.

    PubMed  CAS  Article  Google Scholar 

  34. Ip, C., Ip, M., Loftus, T., Shoemaker, S., and Shea-Eaton, W. (2000) Induction of Apoptosis by Conjugated Linoleic Acid in Cultured Mammary Tumor Cells and Premalignant Lesions of the Rat Mammary Gland, Cancer Epidemiol. Biomarkers Prev. 9, 689–696.

    PubMed  CAS  Google Scholar 

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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

Keywords

  • Conjugate Linoleic Acid
  • Neutral Lipid Fraction
  • Fatty Acid Treatment
  • Conjugate Linoleic Acid Treatment
  • Conjugated Linoleic Acid Level