Effect of CLA (Conjugated Linoleic Acid) on the Anti-Atherosclerotic actors in Human Hepatoma HepG2 Cells

간암세포 (HepG2 Cell)에서의 식이성 CLA(Conjugated Linoleic Acid)가 항동맥경화성 인자에 미치는 영향

  • 오현희 (성신여자대학교 식품영양학과) ;
  • 문희정 (성신여자대학교 식품영양학) ;
  • 이명숙 (성신여자대학교 식품영양학과)
  • Published : 2004.04.01

Abstract

Conjugated linoleic acid (CLA) is the mixture of positional and geometric isomers of linoleic acid (LA), which is found abundantly in dairy products and meats. This study was performed to investigate the anticarcinogenic effect of CLA in HepG2 hepatoma cells. HepG2 cell were treated with LA and CLA at the various concentrations of 10, 20, 40, 80 uM each at different incubation times. After each incubation times, cell proliferation, fatty acids incorporation into cell, peroxidation and postaglandin E$_2$ (PGE$_2$) and thromboxane $A_2$ (TXA$_2$) for the eicosanoid metabolism were measured. LA treated HepG2 cells were increased cell growth 6 - 70% of control whereas CLA increased cell death the half of those in LA group (p 〈 0.001). LA and CLA were incorporated very well into the cellular membranes four times higher than in control according to concentration and longer incubation times. Moreover, LA synthesized significantly arachidonic acids corresponding with LA concentration compared to CLA supplementation. The supplementation with LA increased intracellular lipid peroxides concentration corresponding with LA concentration and five times higher than those in CLA significantly at any incubation times (p 〈 0.001). PGE$_2$ and TXA$_2$ levels were three to twenty times lower in condition of CLA treatments than LA, respectively. Overall, the dietary CLA might change the HepG2 cell growth by the changes of cell composition, production of lipid peroxide. Since CLA have not changed the levels of arachidonic acid of cell membrane, which was sources of eicosanoids, eicosanoid synthesis was not increased in CLA compared to LA. Our results was suggest CLA has a possibility to protect the progress of atherosclerosis because CLA does not produce lipid production and endothelial contraction factors in liver.

Keywords

References

  1. Alcohol Clin Exp Res v.22 Korean status of alcolouics and alcohol related health problems Park,S.C.;Oh,S.I.;Lee,M.S.
  2. Causes of death startistics yearbook National Statistical Office
  3. Arterioscler Thromb v.14 no.4 Dietary polyunsaturated fat decreases coronary artery atherosclerosis in a pediatric-aged population of African green monkeys Wolfe,M.S.;Sawyer,J.K.;Morgan,T.M.;Bullock,B.D.;Rudel.L.L. https://doi.org/10.1161/01.ATV.14.4.587
  4. Am J Clin Nutr v.62 no.2 Dietary polyunsaturated fat modifies low-density lipoproteins and reduces atherosclerosis of non human primates with high and low diet responsiveness Rudel,L.L.;Johnson,F.L.;Sawyer,J.K.;Wilson,M.S.;Parks,J.S. https://doi.org/10.1093/ajcn/62.2.463S
  5. Br Med J v.2 no.6098 Diet and heart: a postscript Morris,J.N.;Marr,J.W.;Clayton,D.G. https://doi.org/10.1136/bmj.2.6098.1307
  6. Arterioscler Throm Vasc Biol v.18 no.11 Dietary monounsaturaated fatty acid promote aortic atherosclerosis in LDL receptor-null, human ApoB100-overexpressing transgenicmice Rudel,L.L.;Kelley,K.;Sawyer,J.K;Shah,R.;Wilson,Md
  7. J Clin Invest v.100 no.1 Hepatic orgin or cholesteryl oleate in cononary artery atherosclerosis in Arican green monkeys. Enrichment by dietary monounsaturated fat Rudel,L.L.;Haines,J.;Sawyer,J.K.;Shah,R.;Wilson,M.S.;Carr,T.P. https://doi.org/10.1172/JCI119524
  8. J Clin Invest v.91 no.2 Effect of oleate-rich and linoleate-rich diets on the susceptibility of low density lipoprotein to oxidative modification in mildly hypercholesterolemic subjects Reaven,P.;Parthasarathy,S.;Grasse,B.J.;Milleer,E.;Steinberg,D.;Witztum,J.L. https://doi.org/10.1172/JCI116247
  9. Biochemistry v.33 no.7 Fatty acid composition of low-density lipoprotein influences its susceptibility to autoxidation Thomas,M.J.;Thoruburg,T.;Manning,J.;Hooper,K.;Rudel,L.L. https://doi.org/10.1021/bi00173a028
  10. First Korean Conference on Science and Technology Anticarcinogenic conjugated derivatives of linoleic acid found in grilled ground beef: isolation, identification, and mechanism of action Ha,Y.L.;Pariza,M.W.
  11. Cancer Res v.45 Requirement of esscential fatty acid for mammary tumorigenesis in the rat Ip,C.;Carter,C.A.;Ip,M.M.
  12. Annu Rev Nutr v.8 Dietary fat and cancer risk: evidence and research needs Pariza,M.W. https://doi.org/10.1146/annurev.nu.08.070188.001123
  13. Cancer Lett v.116 no.2 The growth inhibitory effect of conjugated linoleic acid on MCF-7 cells is released to estrogen response system Durgam,V.R.;Fernandes,G. https://doi.org/10.1016/S0304-3835(97)00192-4
  14. Anticancer Res v.17 no.1A Proliferative responses of normal human mammary and MCF-7 breast cancer cells to linoleic acid, conjugated linoleic acid and eicosanoid synthesis inhibitors in culture Cunningham,D.C.;Harrison,L.Y.;Shultz,T.D.
  15. Carcinogenesis v.8 no.12 Anticarcinogens from fried ground beef: heat-altered derivatives of linoleic acid Ha,Y.L.;Grimm,N.K.;Pariza,M.W. https://doi.org/10.1093/carcin/8.12.1881
  16. Lipid v.30 no.7 Reinvestigation of the antioxidant properties of conjugated linoleic acid Van den Berg,J.J.;Cook,N.E.;Tribble,D.L. https://doi.org/10.1007/BF02536996
  17. J Am Oil Chem Soc v.75 Conjugated linoleic acid and oxidative stress Banni,S.;Angioni,E.;Contini,M.S.;Carta,G.;Casu,V.;Iengo,G.A.;Melis,M.P.;Dians,M.;Dessi,M.A.;Corongiu,F.P. https://doi.org/10.1007/s11746-998-0040-9
  18. Proc Natl Acad Sci USA v.91 no.6 Inducible isoforms of cyclooxygenase and nitric oxide synthase in inflammation Vane,J.R.;Mitchell,J.A.;Appleton,I.;Tomlinson,A.;Bailey,D.B.;Croxtall,J.;Willoughby,D.A. https://doi.org/10.1073/pnas.91.6.2046
  19. Cancer Letters v.127 Conjugated linoleic acid reduces arachidonic acid content and PGE2 synthesis in murine keratin ocytes Kai-Li Liu;Martha A. Belury https://doi.org/10.1016/S0304-3835(97)00479-5
  20. Apolipoprotein Metabolism Lee,M.S.
  21. Artery v.22 no.5 Dietary conjugated linoleic acid reduces plasma lipoproteins and ealy aotic atherosclerosis in hypercholesterolemic hamsters Nicolosi,R.J.;Rogers,E.J.;Kritchevsky,D.;Scimeca,J.A.;Huth,P.J.
  22. Atherosclerosis v.108 no.1 Conjugated linoleic acid and atherosclerosis in rabbits Lee,K.N.;Kritchevsky,D.;Pariza,M.W. https://doi.org/10.1016/0021-9150(94)90034-5
  23. Br J Nutr v.81 no.3 Dietary conjugated linoleic acids promote fatty streak formation in the C57BL/6 mouse atherosclosis model Munday,J.S.;Thompson,K.G.;James,K.A. https://doi.org/10.1017/S0007114599000458
  24. J Lipid Res v.36 no.1 Palmitic acid and linoleic acid metabolism in Caco-2 cells: different triglyceride synthesis and lipoprotein secretion van Greevenbroek,M.M.;Voorhout,W.F.;Erkelens,D.W.;van Meer,G.;de Bruin,T.W.
  25. Cancer Res v.48 no.17 Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines Scudiero,D.A.;Shoemaker,R.H.;Paull,K.D.;Monks,A.;Tierney,S.;Nofziger,T.H.;Currens,M.J.;Seniff,D.;Boyd,M.R.
  26. J Biol Chem v.226 no.1 A simple method for the isolation and punification of total lipids from animal tissues Folch,J.;Lees,M.;Sloane Stanley,G.H.
  27. Anal Chem v.71 Detection of Oxidized Low-Density Lipoproteins.Using Surface Plasmon Responce Katharina,G.;Elizabeth,A.H.H. https://doi.org/10.1021/ac981219n
  28. J Korean Soc Fooc Sci Nutr v.26 no.4 Effect of different sources of dietary n-3 fatty acids on immune response and eicosanoids production Kim,W.K.
  29. J Korean Soc Fooc Nutr v.27 no.1 Activities of phospholipase A2 and cyclooxygenase, and syntheses of thromboxane and prostacyclin in streptozotocin induced diabetic rats Yang,J.A.;Kim,S.O.;Choi,J.H.;Kwag,O.G.;Rhee,S.J.;Chang,H.W.
  30. Anal Biochem v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford,M.M. https://doi.org/10.1016/0003-2697(76)90527-3
  31. Cancer Res v.50 Inhibition of benzo(a)pyreneinduced mouse forestomach neoplasia by conjugated dienoic derivatives of linoleic acid Ha,Y.L.;Storkson,J.;Pariza,M.W.
  32. Biochim Biophys Acta v.1530 The Growth inhibitory effect of conjugated linoleic acid on a human hepatoma cell line, HepG2, is induced by a change in fatty acid metabolism, but not the facilitation of lipid peroxidation in the cells Igrashi,M.;Miyazawa,T. https://doi.org/10.1016/S1388-1981(00)00180-3
  33. Lipids v.30 Reinvestigation of the antioxidant properties of conjugated linoleic acid van den Berg,J.J.;Cook,N.E.;Tribble,D.L. https://doi.org/10.1007/BF02536996
  34. Eur J Cancer v.32A Role of oxygen free radicals in cancer development Dreher,D.;Junod,A.F.
  35. Present knowledge in nutrition(7th Edition) Ekhar,E.Z.;L.J.Filer,Jr.
  36. Biochim Biophys Acta v.1345 Metabolites of conjugated isomers of linoleic acid (CLA) in the rat Sebedio,J.L.;Juaneda,P.;Dobson,G.;Ramilison,I.;Martin,J.C.;Chardigny,J.M.;Christie,W.W. https://doi.org/10.1016/S0005-2760(97)00015-5
  37. Nutrition and Cancer v.33 no.2 Effect of dietary conjugated linoleic acid on phorbol ester-induced PGE2 production and hyperplasia in mouse epidermis Kavanaugh,C.J.;Liu,K.L.;Belury,M.A. https://doi.org/10.1207/S15327914NC330203
  38. Biochim Biophys Acta v.1438 no.2 Antiplatelet effects of conjugated linoleic acid isomers Truitt,A.;McNeil,G.;Vanderhoek,J.Y. https://doi.org/10.1016/S1388-1981(99)00055-4
  39. Lipid v.23 Eicosanoid synthesis in 7,12-dimethylbenz(a) anthracene-induced mammary adenocarci-nomas in Sprague-Dawley rats fed primrose oil, menhaden oil or corn oil diet Abou-EI-Ela,S.H.;Prasse,K.W.;Carroll,R.;Wade,A.E.;Dharwadkar,S.;Bounce,O.R. https://doi.org/10.1007/BF02536342