The Effect of Conjugated Linoleic Acid Isomers on the Cell Proliferation, Apotosis and Expressions of Uncoupling Protein (Ucp) Genes during Differentiation of 3T3-L1 Preadipocytes

Conjugated Linoleic Acid 이성체가 3T3-L1 지방전구세포 분화중 세포증식, 세포사멸 및 Ucp 유전자 발현에 미치는 영향

  • 권소영 (덕성여자대학교 자연과학대학 식품영양학과) ;
  • 강금지 (덕성여자대학교 자연과학대학 식품영양학과)
  • Published : 2004.09.01

Abstract

It has been reported that CLA decreases fat deposition in vivo and in vitro experiments. Among CLA isomers, c9t11 and t10c12 have been shown to exert active biological activities. For example, t10c12 reduces body weight and increases lean body mass, whereas, c9t11 has little effect on body fattness. However, the underlying molecular mechanism for the anti-obesity action of CLA isomers are not well understood. The purpose of this study was to examine the effects of t10c12 and c9t11 on lipid accumulation, cell proliferation, cell death and the expression levels of Ucp genes which are proposed as targets for anti-obesity in 3T3-L1 preadipocytes. Isomers of CLA at 50$\mu$M were added into preadipocyte differentiation medium for 3, 6 and 9days. Control cells received only the vehicle in the differentiation medium. Cytochemical analyses for lipid accumulation, cell proliferation and apotosis were carried out to compare lipidogenesis and cellular activity. RT-PCR analysis of GAPDH, Ucp 2,3 and 4 were also performed to find any modulatory effects of CLA isomers on the metabolic genes. Lipid accumulation indicated by Oil Red-O staining was inhibited in CLA isomers as compared to the control. T10c12 isomer showed less lipidogenesis than c9t11 did. A decrease occurred in CLA isomers as shown by BrdU incorporation. Apotosis has occured at higher level in t10c12 when compared to that of t9c11. Ucp 2, 3 and 4 genes were also upregulated in CLA isomers. T10c12 showed higher level of Ucp gene expressions than the c9t11 did. The biological activities of CLA isomers were also found to be different during differentiation of 3T3-L1 preadipocytes, suggesting that different isomers may be active in certain stage of lipidogenesis. The results indicate that both c9t11 and t10c12 CLA isomers decrease lipidogenesis, inhibit cell proliferation, increase cell death and upregulate in Ucp gene expressions during 3T3-L1 preadipocyte differentiation. T10c12 isomer was more effective than c9t11 in overall anti-obesity activity.

Keywords

References

  1. Pariza MW, Park YH, Cook ME. The biologically active isomers of conjugated linoleic acid. Prog Lipid Res 40: 283-298, 2001 https://doi.org/10.1016/S0163-7827(01)00008-X
  2. Evans ME, Brown JM, McIntosh MK. Isomer-specific effects of conjugated linoleic acid (CLA) on adipocity and lipid metabolism. J Nutrbiochem 13 (9): 508-516, 2002
  3. Pariza MW, Park YH, Cook ME. Mechanism of action of conjugated linoleic acid: evidence and speculation. Proc Soc Exp Bioi Med 223: 8-13, 2000 https://doi.org/10.1046/j.1525-1373.2000.22302.x
  4. Belury MA. Dietary conjugated linoleic acid in health: physiological effects and mechanisms of action. Annu Rev Nutr 22: 505-531, 2002 https://doi.org/10.1146/annurev.nutr.22.021302.121842
  5. Haumann BE. Conjugated linoleic acid. Inform 7 (2): 152-159, 1996
  6. Delany JP, West DB. Changes in body composition with conjugated linoleic acid. Am Coll Nutr 19 (4): 487s-493s, 2000 https://doi.org/10.1080/07315724.2000.10718952
  7. Tsuboyama-Kassoka N, Takahashi M, Tanemura K, Kim HJ, Tange T, Okuyarna H, Kasai M, Ikemoto S, Ezaki O. Conjugated linoleic acid supplementation reduces adipose tissue by apotosis and develops lipodystrophy in mice. Diabetes 49: 1534-1542, 2000 https://doi.org/10.2337/diabetes.49.9.1534
  8. Park YH, Storkson JM, Albright KJ, Liy W, Pariza MW. Evidence that the trans-10, cis-12 isomer of conjugated linoleic acid induces body composition changes in mice. Lipids 34(3): 235-241, 1999 https://doi.org/10.1007/s11745-999-0358-8
  9. Evans M, Geigerman C, Cook J, Curtis L, Kuebler B, McIntosh M. Conjugated linoleic acid suppresses triglyceride accumulation and induces apoptosis in 3T3-Ll preadipocytes. Lipids 35: 899-910,2000 https://doi.org/10.1007/S11745-000-0599-6
  10. Evans M, Lin X, OdIe J, McIntosh M. Trans-10, cis-12 conjugated linoleic acid increases fatty acid oxidation in 3T3-Ll preadipocytes. J Nutr 132: 450-455, 2002
  11. Han SA. Effects of conjugated linoleic acid (CLA) on 3T3-L1 preadipocyte differentiation. Master thesis, Chungbuk National Univ,2003
  12. McNeel RL, Mersmann HJ. Effects of isomers of CLA on porcine adipocyte growth and differentiation. J Nutr Biochem 14: 266-274, 2003 https://doi.org/10.1016/S0955-2863(03)00031-7
  13. Belury MA, Kempa-Steczko A. Conjugated linoleic acid modulates hepatic lipid composition in mice. Lipids 32: 199-204, 1997 https://doi.org/10.1007/s11745-997-0025-0
  14. Sisk M, Azain MI, Hausman DB, Jewell DE. Effect of conjugated linoleic acid on fat pad weights and celluarity in Sprague Dawley and Zucker rats. FASEB (abstract) 12: A536, 1998
  15. Satory DL, Smith SB. Conjugated linoleic acid inhibits proliferation but stimulates lipid filling of murine 3T3-L1 preadipocytes. J Nutr 129: 92-97, 1999
  16. Brodie AE, Manning VA, Ferguson KR, Jewell DE, Hu CY. Conjugated linoleic acid inhibits differentiation of pre- and post- con-fluent 3T3-Ll preadipocytes but inhibits cell proliferation only in preconfluent cells. J Nutr 129: 602-606, 1999
  17. Takahashi Y, Kushiro M, Shinohara K, Ide K. Dietary conjugated linoleic acid reduces body fat mass and affects gene expression of proteins regulating energy metabolism in mice. Comp Biochem Physiol 133 (3) : 395-404, 2002 https://doi.org/10.1016/S1096-4959(02)00164-1
  18. Kang KH, Liu W, Albright KJ, Park YH, Pariza MW. Trans-10, cis-12 CLA inhibits differentiation of 3T3-Ll adipocytes and decreased PPAR \gamma expression. Biochem Biophys Res Commun 303 (3): 795-799, 2003 https://doi.org/10.1016/S0006-291X(03)00413-3
  19. Choi Y, Kim YC, Han YB, Park Y, Pariza MW, Ntami JM. The trans-10, cis-10 isomer of conjugated linoleic acid downregulates stearoyl-Co A desaturase 1 gene expression in 3T3-Ll adipocytes. J Nutr 130: 1920-1924, 2000
  20. Palou A, Serra F, bonet ML, Pico C. Obesity: molecular bases of a multifactorial problem. Eur J Nutr 39: 127-144,2000 https://doi.org/10.1007/s003940070017
  21. Jezek P. Possible physiological roles of mitochondrial uncoupling proteins-UCPn. Int J Biochem Cell Biol 34: 1190-1206, 2002 https://doi.org/10.1016/S1357-2725(02)00061-4
  22. Rodriguez E, Ribot J, Palou A. Trans-10, cis-12, but not cis-9, trans-11 CLA isomer, inhibits brown adipocyte thermogenic capacity. Am J Physiol 282 (6) : R1787-1797, 2002
  23. West DB, Blohm FY, Truett AA, Delany JP. Conjugated linoleic acid persistently increases total energy expenditure in AKR/J mice without increasing uncoupling protein gene expression. J Nutr 130(10): 2471-2477, 2000
  24. Richter c, Schweizer M, cossarizza A, Franceschi C. Control of apotosis by the cellular ATP level. FEBS Lett 378: 107-110,1996 https://doi.org/10.1016/0014-5793(95)01431-4
  25. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156-159, 1987
  26. Kang KJ, Kwon SY. Effects of wax gourd extracts on adipocyted differentiation and uncoupling gene (Ucp) expressions in 3T3-Ll preadipocytes. Nutr Sci 6: 148-154, 2003
  27. Brown JM, Halvorsen YD, Lea-currie YR, Geigerman C, McIntosh M. Trans-10, cis-12. but not cis-9, trans-11, conjugated linoleic acid attenuates lipogenesis in primary cultures of stromal vascular cells from human adipose tissue. J Nutr 131: 2316-2321. 2001
  28. Hensey C, Gautier J. A developmental timer that regulates apoptosis at the onset of gastrulation. Mech Dev 69: 183-195, 1997 https://doi.org/10.1016/S0925-4773(97)00191-3
  29. Ealey KN, EI-Sohemy A, Archer MC. Effects of dietary conjugated linoleic acid on the expression of uncoupling proteins in mice and rats. Lipids 37 (9): 853-861, 2002 https://doi.org/10.1007/s11745-002-0971-6
  30. Roche HM, Noone E, Sewter C, Mc Bennett S, Savage D, Gibney MJ, O'Rahilly S, Vidal-Puig AI. Isomer-dependent metabolic effects of conjugated linoleic acid: insights from molecular markers sterol regulatory element-binding protein-1c and LXRapha. Diabetes 51 (7): 2037-2044, 2002 https://doi.org/10.2337/diabetes.51.7.2037
  31. Dulloo AG, Samec S. Uncoupling proteins: their roles in adaptive thermogenesis and substrate metabolism reconsidered. Br J Nutr 86: 123-139, 2001 https://doi.org/10.1079/BJN2001412