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Conjugated Linoleic Acids and CLA-Containing Phospholipids Inhibit NO Formation in Aortic Endothelial Cells

  • Original Article
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Lipids

Abstract

This study examined the effects of five CLA isomers and the non-conjugated LA on nitric oxide (NO) production, an important modulator of vasodilation, inflammation, and cytotoxicity. Bovine aortic endothelial cells (BAECs) were pretreated with pure CLAs (c9, c11-, c9, t11-, t9, t11-, t10, c12-, c11, c13-CLA) and the non-conjugated c9, c12-analog, then stimulated by the ionophore A23187 followed by fluorescence monitoring of NO production. CLAs (5 μM) decreased NO formation in the range of 20–40% relative to non-fatty acid-treated controls with the t9, t11- and t10, c12-CLAs being the most effective. The inhibitory effect of either of these CLAs was not time dependent over a 120 min time interval. Phosphatidylcholine (PC) and phosphatidylethanolamine (PE) play crucial roles in membrane structure and cell signaling. Since CLAs are usually esterified in these phospholipids (PLs), the effects of three CLA-containing PLs and 2-linoleoyl-PC on NO production by A23187-stimulated BAECs were also examined. c9, t11-CLA-PC and 2-linoleoyl-PC dose-dependently inhibited NO production over a 60–1,000 μM concentration range whereas c9, c11-CLA-PC and c9, t11-CLA-PE were ineffective. Both c9, t11-CLA-PC and linoleoyl-PC exhibited a time-dependent decrease in NO production from 5 to 120 min. The results of the present study suggest that the influence of several C18 polyunsaturated fatty acids incorporated into cellular phospholipids on the A23187-induced formation of NO by BAECs is strongly dependent on the positional and geometric nature of the double bonds.

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Abbreviations

BAEC:

Bovine aortic endothelial cells

CLA:

Conjugated linoleic acid

DAN:

2, 3-Diaminonaphthalene

ECGS:

Endothelial cell growth supplement

FBS:

Fetal bovine serum

FU:

Fluorescence units

L-Gln:

l-Glutamine

HBSS:

Hank’s balanced salt solution

NO:

Nitric oxide

NOS:

Nitric oxide synthase

c9, c11-CLA-PC:

1-Stearoyl-2-(c9, c11-octadecadienoyl)-sn-glycerol-3-phosphocholine

linoleoyl-PC:

1-Stearoyl-2-linoleoyl-sn-glycerol-3-phosphocholine

c9, t11-CLA-PC:

1-Stearoyl-2-(c9, t11-octadecadienoyl)-sn-glycerol-3-phosphocholine

c9, t11-CLA-PE:

1-Stearoyl-2-(c9, t11-octadecadienoyl)-sn-glycerol-3-phosphoethanolamine

PL:

Phospholipid

References

  1. Rastaldo R, Paglioro P, Cappello S, Penna C, Mancardi D, Westerhof N, Losano G (2007) Nitric oxide and cardiac function. Life Sci 81:779–793

    Article  PubMed  CAS  Google Scholar 

  2. Wiley JW (2007) The many faces of nitric oxide: cytotoxic, cytoprotective or both. Neurogastroenterol Motil 19:541–544

    Article  PubMed  CAS  Google Scholar 

  3. Mariotto S, Suzuki Y, Persichini T, Colasanti M, Suzuki H, Cantoni O (2007) Cross-talk between NO and arachidonic acid in inflammation. Curr Med Chem 14:1940–1944

    Article  PubMed  CAS  Google Scholar 

  4. Fleming I, Busse R (2003) Molecular mechanisms involved in the regulation of the endothelial nitric oxide synthase. Am J Physiol Regul Integr Comp Physiol 284:R1–R12

    PubMed  CAS  Google Scholar 

  5. Nathan C, Xie Q-W (1994) Nitric oxide synthases: roles, tolls and controls. Cell 78:915–918

    Article  PubMed  CAS  Google Scholar 

  6. Fulton D, Gratton J-P, Sessa WC (2001) Post-translational control of endothelial nitric oxide synthase: why isn’t calcium/calmodulin enough? J Pharm Exp Therap 299:818–824

    CAS  Google Scholar 

  7. Kimura C, Oike M, Ohnaka K, Nose Y, Ito Y (2003) Constitutive nitric oxide production in bovine aortic and brain microvascular endothelial cells: a comparative study. J Physiol 554:721–730

    Article  PubMed  Google Scholar 

  8. Miles PR, bowman L, Rengasamy A, Huffman L (1998) Constitutive nitric oxide production by rat alveolar macrophages. Am J Physiol 274:L360–L368

    PubMed  CAS  Google Scholar 

  9. Shen L-J, Lin W-C, Beloussow K, Hosoya K-I, Terasaki T, Ann DK, Shen W-C (2003) Recombinant arginine deiminase as a differential modulator of inducible (iNOS) and endothelial (eNOS) nitric oxide synthetase activity in cultured endothelial cells. Biochem Pharmacol 66:1945–1952

    Article  PubMed  CAS  Google Scholar 

  10. Korhone R, Kankaanranta H, Lahti A, Lahde M, Knowles RG, Moilanen E (2001) Bi-directional effects of the elevation of intracellular calcium on the expression of inducible nitric oxide synthase in J774 macrophages exposed to low and to high concentrations of endotoxin. Biochem J 354:351–358

    Article  Google Scholar 

  11. Ha YL, Storkson J, Pariza MW (1990) Inhibition of benzo(a)pyrene-induced mouse forestomach neoplasia by conjugated dienoic derivatives of linoleic acid. Cancer Res 50:1097–1101

    PubMed  CAS  Google Scholar 

  12. Al-Madaney MM, Kramer JKG, Deng Z, Vanderhoek JY (2003) Effects of lipid-esterified isomers on platelet function: evidence for stimulation of platelet phospholipase activity. Biochim Biophys Acta 1635:75–82

    PubMed  CAS  Google Scholar 

  13. Coen PM, Cummins PM, Birney YA, Devery R, Cahill PA (2004) Modulation of nitric oxide and 6-keto-prostaglandin F1α production in bovine aortic endothelial cells by conjugated linoleic acid. Endothelium 11:211–220

    Article  PubMed  CAS  Google Scholar 

  14. Eder K, Schleser S, Becker K, Körting R (2003) Conjugated linoleic acids lower the release of eicosanoids and nitric oxide from human aortic endothelial cells. J Nutr 133:4083–4089

    PubMed  CAS  Google Scholar 

  15. Iwakiri Y, Sampson DA, Allen KGD (2002) Suppression of cyclooxygenase-2 and inducible nitric oxide synthase expression by conjugated linoleic acid in murine macrophages. Prostaglandins, Leukot Essent Fatty Acids 67:435–443

    Article  CAS  Google Scholar 

  16. Sheu JN, Lin TH, Lii CK, Chen CC, Chen HW, Liu KL (2006) Contribution of conjugated linoleic acid to the suppression of inducible nitric oxide synthase expression and transcription factor activation in stimulated mouse mesangial cells. Food Chem Toxicol 44:409–416

    Article  PubMed  CAS  Google Scholar 

  17. Ohashi Y, Katayama M, Hirata K, Suematsu M, Kawashima S, Yokoyama M (1993) Activation of nitric oxide synthase from cultured aortic endothelial cells by phospholipids. Biochem Biophys Res Commun 195(3):1314–1320

    Article  PubMed  CAS  Google Scholar 

  18. Lai KL, Torres-Duarte AP, Vanderhoek JY (2005) 9-Trans, 11-trans: antiproliferative and proapoptotic effects on bovine endothelial cells. Lipids 40:1107–1116

    Article  PubMed  CAS  Google Scholar 

  19. Rathel T, Leikert JF, Vollmar AM, Dirsch VM (2003) Application of 4,5-diaminofluorescein to reliably measure nitric oxide released from endothelial cells in vitro. Biol Proced Online 5:136–142

    Article  PubMed  CAS  Google Scholar 

  20. Misko TP, Schilling RJ, Salvemini D, Moore WM, Currie MG (1993) A fluorometric assay for the measurement of nitrite in biological samples. Anal Biochem 214:11–16

    Article  PubMed  CAS  Google Scholar 

  21. Meininger C, Wu G (1997) l-glutamine inhibits nitric oxide synthesis in bovine venular endothelial cells. J Pharmacol and Exp Ther 281:448–453

    CAS  Google Scholar 

  22. Shen CL, Dunn DM, Henry JH, Yong L, Watkins BA (2004) Decreased production of inflammatory mediators in human osteoarthritic chondrocytes by conjugated linoleic acids. Lipids 39:161–166

    PubMed  CAS  Google Scholar 

  23. Fogerty AC, Ford GL, Svoronos D (1988) Octa-9, 11-dienoic acid in foodstuffs and in the lipids of human blood and breast milk. Nutr Rep Intl 38:937–944

    CAS  Google Scholar 

  24. Huang Y-C, Luedecke LO, Shultz TD (1994) Effect of cheddar cheese consumption on plasma conjugated linoleic acid concentrations in men. Nutr Res 14:373–386

    Article  CAS  Google Scholar 

  25. Torres-Duarte AP, Vanderhoek JY (2003) Conjugated linoleic acid exhibits stimulatory and inhibitory effects on prostanoid production in human endothelial cells and platelets. Biochim Biophys Acta 1640:69–76

    Article  PubMed  CAS  Google Scholar 

  26. Li Q, Zhang Q, Wang M, Zhao S, Ma J, Luo N, Li N, Li Y, Xu G, Li J (2007) Eicosapentaenoic acid modifies lipid composition in caveolae and induces translocation of endothelial nitric oxide synthase. Biochimie 89:169–177

    Article  PubMed  Google Scholar 

  27. Li Q, Zhang Q, Wang M, Liu F, Zhao S, Ma J, Luo N, Li N, Li Y, Xu G, Li J (2007) Docosahexaenoic acid affects endothelial nitric oxide synthase in caveolae. Arch Biochem Biophys 466:250–259

    Article  PubMed  CAS  Google Scholar 

  28. Hong SL, McLaughlin NJ, Tzeng CY, Patton G (1985) Prostacyclin synthesis and deacylation of phospholipids in human endothelial cells: comparison of thrombin, histamine and ionophore A23187. Thromb Res 38:1–10

    Article  PubMed  CAS  Google Scholar 

  29. Niwano K, Arai M, Tomaru K, Uchiyama T, Ohyama Y, Kurabayashi M (2003) Transcriptional stimulation of the eNOS gene by the stable prostacyclin analogue breaprots is mediated through cAMP-responsive element in vascular endothelial cells. Close link between PGI2 signal and NO pathways. Circ Res 93:523–530

    Article  PubMed  CAS  Google Scholar 

  30. Stachowska E, Dziedziejko V, Safranow K, Gutowska I, Adler G, Ciechanowicz A, Machalinski B, Chlubek D (2007) Inhibition of phospholipase A2 activity by conjugated linoleic acids in human macrophages. Eur J Nutr 46:28–33

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, The George Washington University School of Medicine, 2300 Eye Street, N.W., Washington, DC, 20037, USA

    Kimberly J. Jenko & Jack Y. Vanderhoek

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  1. Kimberly J. Jenko
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  2. Jack Y. Vanderhoek
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Correspondence to Jack Y. Vanderhoek.

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Jenko, K.J., Vanderhoek, J.Y. Conjugated Linoleic Acids and CLA-Containing Phospholipids Inhibit NO Formation in Aortic Endothelial Cells. Lipids 43, 335–342 (2008). https://doi.org/10.1007/s11745-008-3160-y

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  • DOI: https://doi.org/10.1007/s11745-008-3160-y

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