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Walnut extract (Juglans regia L.) and its component ellagic acid exhibit anti-inflammatory activity in human aorta endothelial cells and osteoblastic activity in the cell line KS483

  • Z. Papoutsi (a1), E. Kassi (a1), I. Chinou (a2), M. Halabalaki (a2), L. A. Skaltsounis (a2) and P. Moutsatsou (a1)...


Epidemiological studies suggest that the incidence of CVD and postmenopausal osteoporosis is low in the Mediterranean area, where herbs and nuts, among others, play an important role in nutrition. In the present study, we sought a role of walnuts (Juglans regia L.) in endothelial and bone-cell function. As the endothelial cell expression of adhesion molecules has been recognised as an early step in inflammation and atherogenesis, we examined the effect of walnut methanolic extract and ellagic acid, one of its major polyphenolic components (as shown by HPLC analysis), on the expression of vascular cell adhesion molecule (VCAM)-1 and intracellular adhesion molecule (ICAM)-1 in human aortic endothelial cells. After incubating the cells with TNF-α (1 ng/ml) in the absence and in the presence of walnut extract (10–200 μg/ml) or ellagic acid (10− 7–10− 5 m), the VCAM-1 and ICAM-1 expression was quantified by cell-ELISA. We further evaluated the effect of walnut extract (10–50 μg/ml), in comparison with ellagic acid (10− 9–10− 6m), on nodule formation in the osteoblastic cell line KS483.Walnut extract and ellagic acid decreased significantly the TNF-α-induced endothelial expression of both VCAM-1 and ICAM-1 (P < 0·01; P < 0·001). Both walnut extract (at 10–25 μg/ml) and ellagic acid (at 10− 9–10− 8 m) induced nodule formation in KS483 osteoblasts. The present results suggest that the walnut extract has a high anti-atherogenic potential and a remarkable osteoblastic activity, an effect mediated, at least in part, by its major component ellagic acid. Such findings implicate the beneficial effect of a walnut-enriched diet on cardioprotection and bone loss.

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      Walnut extract (Juglans regia L.) and its component ellagic acid exhibit anti-inflammatory activity in human aorta endothelial cells and osteoblastic activity in the cell line KS483
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Corresponding author

*Corresponding author: Dr P. Moutsatsou, fax +30 210 7462682, email


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1 de Lorgeril, M & Salen, P (2006) The Mediterranean-style diet for the prevention of cardiovascular diseases. Public Health Nutr 9, 118123.
2 Kris-Etherton, P, Eckel, RH, Howard, BV, St Jeor, S & Bazzarre, TL (2001) AHA Science Advisory: Lyon Diet Heart Study. Benefits of a Mediterranean-style, National Cholesterol Education Program/American Heart Association Step I dietary pattern on cardiovascular disease. Circulation 103, 18231825.
3 Fogarty, P, O'Beirne, B & Casey, C (2005) Epidemiology of the most frequent diseases in the European a-symptomatic post-menopausal women. Is there any difference between Ireland and the rest of Europe? Maturitas 52, Suppl. 1, S3S6.
4 Carluccio, MA, Siculella, L, Ancora, MA, Massaro, M, Scoditti, E, Storelli, C, Visioli, F, Distante, A & De Caterina, R (2003) Olive oil and red wine antioxidant polyphenols inhibit endothelial activation: antiatherogenic properties of Mediterranean diet phytochemicals. Arterioscler Thromb Vasc Biol 23, 622629.
5 Gebauer, SK, Psota, TL, Harris, WS & Kris-Etherton, PM (2006) n-3 Fatty acid dietary recommendations and food sources to achieve essentiality and cardiovascular benefits. Am J Clin Nutr 83, 1526S1535S.
6 Kris-Etherton, PM, Hecker, KD, Bonanome, A, Coval, SM, Binkoski, AE, Hilpert, KF, Griel, AE & Etherton, TD (2002) Bioactive compounds in foods: their role in the prevention of cardiovascular disease and cancer. Am J Med 113, Suppl. 9B, 71S88S.
7 Maguire, LS, O'Sullivan, SM, Galvin, K, O'Connor, TP & O'Brien, NM (2004) Fatty acid profile, tocopherol, squalene and phytosterol content of walnuts, almonds, peanuts, hazelnuts and the macadamia nut. Int J Food Sci Nutr 55, 171178.
8 Fukuda, T, Ito, H & Yoshida, T (2003) Antioxidative polyphenols from walnuts (Juglans regia L.). Phytochemistry 63, 795801.
9 Almario, RU, Vonghavaravat, V, Wong, R & Kasim-Karakas, SE (2001) Effects of walnut consumption on plasma fatty acids and lipoproteins in combined hyperlipidemia. Am J Clin Nutr 74, 7279.
10 Zambon, D, Sabate, J, Munoz, S, Campero, B, Casals, E, Merlos, M, Laguna, JC & Ros, E (2000) Substituting walnuts for monounsaturated fat improves the serum lipid profile of hypercholesterolemic men and women. A randomized crossover trial. Ann Intern Med 132, 538546.
11 Ros, E, Nunez, I, Perez-Heras, A, Serra, M, Gilabert, R, Casals, E & Deulofeu, R (2004) A walnut diet improves endothelial function in hypercholesterolemic subjects: a randomized crossover trial. Circulation 109, 16091614.
12 Wu, D, Koga, T, Martin, KR & Meydani, M (1999) Effect of vitamin E on human aortic endothelial cell production of chemokines and adhesion to monocytes. Atherosclerosis 147, 297307.
13 Anderson, KJ, Teuber, SS, Gobeille, A, Cremin, P, Waterhouse, AL & Steinberg, FM (2001) Walnut polyphenolics inhibit in vitro human plasma and LDL oxidation. J Nutr 131, 28372842.
14 Festa, F, Aglitti, T, Duranti, G, Ricordy, R, Perticone, P & Cozzi, R (2001) Strong antioxidant activity of ellagic acid in mammalian cells in vitro revealed by the comet assay. Anticancer Res 21, 39033908.
15 Mertens-Talcott, SU, Talcott, ST & Percival, SS (2003) Low concentrations of quercetin and ellagic acid synergistically influence proliferation, cytotoxicity and apoptosis in MOLT-4 human leukemia cells. J Nutr 133, 26692674.
16 Han, SH, Quon, MJ & Koh, KK (2007) Reciprocal relationships between abnormal metabolic parameters and endothelial dysfunction. Curr Opin Lipidol 18, 5865.
17 McFarlane, SI, Muniyappa, R, Shin, JJ, Bahtiyar, G & Sowers, JR (2004) Osteoporosis and cardiovascular disease: brittle bones and boned arteries, is there a link? Endocrine 23, 110.
18 Park, YH, Han, DW, Suh, H, Ryu, GH, Hyon, SH, Cho, BK & Park, JC (2003) Protective effects of green tea polyphenol against reactive oxygen species-induced oxidative stress in cultured rat calvarial osteoblast. Cell Biol Toxicol 19, 325337.
19 Puel, C, Quintin, A, Mathey, J, Obled, C, Davicco, MJ, Lebecque, P, Kati-Coulibaly, S, Horcajada, MN & Coxam, V (2005) Prevention of bone loss by phloridzin, an apple polyphenol, in ovariectomized rats under inflammation conditions. Calcif Tissue Int 77, 311318.
20 Griel, AE, Kris-Etherton, PM, Hilpert, KF, Zhao, G, West, SG & Corwin, RL (2007) An increase in dietary n-3 fatty acids decreases a marker of bone resorption in humans. Nutr J 6, 2.
21 Hsu, YL, Chang, JK, Tsai, CH, Chien, TT & Kuo, PL (2007) Myricetin induces human osteoblast differentiation through bone morphogenetic protein-2/p38 mitogen-activated protein kinase pathway. Biochem Pharmacol 73, 504514.
22 Kuo, PL (2005) Myricetin inhibits the induction of anti-Fas IgM-, tumor necrosis factor-α- and interleukin-1β-mediated apoptosis by Fas pathway inhibition in human osteoblastic cell line MG-63. Life Sci 77, 29642976.
23 Bai, XC, Lu, D, Bai, J, Zheng, H, Ke, ZY, Li, XM & Luo, SQ (2004) Oxidative stress inhibits osteoblastic differentiation of bone cells by ERK and NF-κB. Biochem Biophys Res Comm 314, 197–207.
24 Tokuda, H, Takai, S, Hanai, Y, Matsushima-Nishiwaki, R, Hosoi, T, Harada, A, Ohta, T & Kozawa, O (2007) (-)-Epigallocatechin gallate suppresses endothelin-1-induced interleukin-6 synthesis in osteoblasts: inhibition of p44/p42 MAP kinase activation. FEBS Lett 581, 13111316.
25 Colaric, M, Veberic, R, Solar, A, Hudina, M & Stampar, F (2005) Phenolic acids, syringaldehyde, and juglone in fruits of different cultivars of Juglans regia L. J Agric Food Chem 53, 63906396.
26 Li, L, Tsao, R, Yang, R, Liu, C, Zhu, H & Young, JC (2006) Polyphenolic profiles and antioxidant activities of heartnut (Juglans ailanthifolia var cordiformis) and Persian walnut (Juglans regia L.). J Agric Food Chem 54, 80338040.
27 Jurd, L (1956) Plant polyphenols. I. The polyphenolic constituents of the pellicle of the walnut (Juglans regia). J. Am Chem Soc 78, 3445–3448.
28 Jurd, L (1958) Plant polyphenols. III. The isolation of a new ellagitannin from the pellicle of the walnut. Am Chem Soc 80, 2249–2252.
29 Stampar, F, Solar, A, Hudina, M, Veberic, R & Colaric, M (2006) Traditional walnut liqueur – cocktail of phenolics. Food Chem 95, 627–631.
30 Jakopic, J, Colaric, M, Veberic, R, Hudina, M, Solar, A & Stampar, F (2007) How much do cultivar and preparation time influence on phenolics content in walnut liqueur? Food Chem 104, 100–105.
31 Papoutsi, Z, Kassi, E, Tsiapara, A, Fokialakis, N, Chrousos, GP & Moutsatsou, P (2005) Evaluation of estrogenic/antiestrogenic activity of ellagic acid via the estrogen receptor subtypes ERα and ERβ. J Agric Food Chem 53, 77157720.
32 Zhang, WJ, Stocker, R, McCall, MR, Forte, TM & Frei, B (2002) Lack of inhibitory effect of HDL on TNFα-induced adhesion molecule expression in human aortic endothelial cells. Atherosclerosis 165, 241–249.
33 Denizot, F & Lang, R (1986) Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods 89, 271277.
34 Tsuda, K & Nishio, I (2004) Modulation of endothelial function by walnuts and sex hormones. Circulation 110, e73.
35 Kaneko, M, Hayashi, J, Saito, I & Miyasaka, N (1996) Probucol downregulates E-selectin expression on cultured human vascular endothelial cells. Arterioscler Thromb Vasc Biol 16, 10471051.
36 Wolle, J, Hill, RR, Ferguson, E, Devall, LJ, Trivedi, BK, Newton, RS & Saxena, U (1996) Selective inhibition of tumor necrosis factor-induced vascular cell adhesion molecule-1 gene expression by a novel flavonoid. Lack of effect on transcription factor NF-κB. Arterioscler Thromb Vasc Biol 16, 15011508.
37 Weber, C, Erl, W, Pietsch, A, Strobel, M, Ziegler-Heitbrock, HW & Weber, PC (1994) Antioxidants inhibit monocyte adhesion by suppressing nuclear factor-κB mobilization and induction of vascular cell adhesion molecule-1 in endothelial cells stimulated to generate radicals. Arterioscler Thromb 14, 16651673.
38 Walgren, RA, Walle, UK & Walle, T (1998) Transport of quercetin and its glucosides across human intestinal epithelial Caco-2 cells. Biochem Pharmacol 55, 17211727.
39 Whitley, AC, Stoner, GD, Darby, MV & Walle, T (2003) Intestinal epithelial cell accumulation of the cancer preventive polyphenol ellagic acid – extensive binding to protein and DNA. Biochem Pharmacol 66, 907–915.
40 de Ancos, B, Gonzalez, EM & Cano, MP (2000) Ellagic acid, vitamin C, and total phenolic contents and radical scavenging capacity affected by freezing and frozen storage in raspberry fruit. J Agric Food Chem 48, 45654570.
41 Sellappan, S, Akoh, CC & Krewer, G (2002) Phenolic compounds and antioxidant capacity of Georgia-grown blueberries and blackberries. J Agric Food Chem 50, 24322438.
42 Seeram, NP, Lee, R & Heber, D (2004) Bioavailability of ellagic acid in human plasma after consumption of ellagitannins from pomegranate (Punica granatum L.) juice. Clin Chim Acta 348, 63–68.
43 de Vries, JH, Hollman, PC, Meyboom, S, Buysman, MN, Zock, PL, van Staveren, WA & Katan, MB (1998) Plasma concentrations and urinary excretion of the antioxidant flavonols quercetin and kaempferol as biomarkers for dietary intake. Am J Clin Nutr 68, 60–65.
44 Wiseman, H (1999) The bioavailability of non-nutrient plant factors: dietary flavonoids and phyto-oestrogens. Proc Nutr Soc 58, 139146.
45 Manach, C, Morand, C, Crespy, V, Demigne, C, Texier, O, Regerat, F & Remesy, C (1998) Quercetin is recovered in human plasma as conjugated derivatives which retain antioxidant properties. FEBS Lett 426, 331336.
46 Janssen, K, Mensink, RP, Cox, FJ, Harryvan, JL, Hovenier, R, Hollman, PC & Katan, MB (1998) Effects of the flavonoids quercetin and apigenin on hemostasis in healthy volunteers: results from an in vitro and a dietary supplement study. Am J Clin Nutr 67, 255262.
47 Harris, DM, Besselink, E, Henning, SM, Go, VL & Heber, D (2005) Phytoestrogens induce differential estrogen receptor α- or β-mediated responses in transfected breast cancer cells. Exp Biol Med (Maywood) 230, 558568.
48 Islam, KN, Devaraj, S & Jialal, I (1998) α-Tocopherol enrichment of monocytes decreases agonist-induced adhesion to human endothelial cells. Circulation 98, 22552261.
49 Ludwig, A, Lorenz, M, Grimbo, N, Steinle, F, Meiners, S, Bartsch, C, Stangl, K, Baumann, G & Stangl, V (2004) The tea flavonoid epigallocatechin-3-gallate reduces cytokine-induced VCAM-1 expression and monocyte adhesion to endothelial cells. Biochem Biophys Res Commun 316, 659665.
50 Zapolska-Downar, D, Zapolski-Downar, A, Markiewski, M, Ciechanowicz, A, Kaczmarczyk, M & Naruszewicz, M (2000) Selective inhibition by α-tocopherol of vascular cell adhesion molecule-1 expression in human vascular endothelial cells. Biochem Biophys Res Commun 274, 609–615.
51 Kassi, E, Papoutsi, Z, Fokialakis, N, Messari, I, Mitakou, S & Moutsatsou, P (2004) Greek plant extracts exhibit selective estrogen receptor modulator (SERM)-like properties. J Agric Food Chem 52, 69566961.
52 Yu, YM, Wang, ZH, Liu, CH & Chen, CS (2007) Ellagic acid inhibits IL-1β-induced cell adhesion molecule expression in human umbilical vein endothelial cells. Br J Nutr 97, 692698.
53 Murase, T, Kume, N, Hase, T, Shibuya, Y, Nishizawa, Y, Tokimitsu, I & Kita, T (1999) Gallates inhibit cytokine-induced nuclear translocation of NF-κB and expression of leukocyte adhesion molecules in vascular endothelial cells. Arterioscler Thromb Vasc Biol 19, 1412–1420.
54 Kim, JD, Liu, L, Guo, W & Meydani, M (2006) Chemical structure of flavonols in relation to modulation of angiogenesis and immune-endothelial cell adhesion. J Nutr Biochem 17, 165–176.
55 Lee, G, Na, HJ, Namkoong, S, Jeong Kwon, H, Han, S, Ha, KS, Kwon, YG, Lee, H & Kim, YM (2006) 4-O-methylgallic acid down-regulates endothelial adhesion molecule expression by inhibiting NF-κB-DNA-binding activity. Eur J Pharmacol 551, 143–151.
56 Zhao, G, Etherton, TD, Martin, KR, West, SG, Gillies, PJ & Kris-Etherton, PM (2004) Dietary α-linolenic acid reduces inflammatory and lipid cardiovascular risk factors in hypercholesterolemic men and women. J Nutr 134, 29912997.
57 Cerda, B, Tomas-Barberan, FA & Espin, JC (2005) Metabolism of antioxidant and chemopreventive ellagitannins from strawberries, raspberries, walnuts, and oak-aged wine in humans: identification of biomarkers and individual variability. J Agric Food Chem 53, 227235.
58 Manolagas, SC, Kousteni, S & Jilka, RL (2002) Sex steroids and bone. Recent Prog Horm Res 57, 385–409.
59 Kousteni, S, Han, L, Chen, JR, Almeida, M, Plotkin, LI, Bellido, T & Manolagas, SC (2003) Kinase-mediated regulation of common transcription factors accounts for the bone-protective effects of sex steroids. J Clin Invest 111, 16511664.


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Walnut extract (Juglans regia L.) and its component ellagic acid exhibit anti-inflammatory activity in human aorta endothelial cells and osteoblastic activity in the cell line KS483

  • Z. Papoutsi (a1), E. Kassi (a1), I. Chinou (a2), M. Halabalaki (a2), L. A. Skaltsounis (a2) and P. Moutsatsou (a1)...


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