E
concentration (μM)
monocytes vehicle 10 20
Mean fluorescence intensity of CD36 (MFI)
0 50 100 150 200 250
DTF NOB
+ PMA (30 nM)
**
(Figure 3E)
vehicle monocytes
20 μM DTF 20 μM NOB
+PMA
F
monocyte Vehicle 10 20
0 2 4 6 8 10 12 14
DTF NOB
Mean flourescence intensity of SR-A (MFI)
concentration (μM)
+ PMA (30 nM)
*
(Figure 3F)
+ PMA 1.6 nM
monocyte vehicle 10 20
CD36 mRNA expression (fold of monocyte)
0.0
monocyte vehicle 10 20
SR-A mRNA expression (fold of monocyte)
0.0
monocyte vehicle 10 20
LOX-1 mRNA expression (fold of monocyte)
0.0
A
B
**
**
*
#
#
concentration (μM)
monocytes vehicle 10 20
24 h DiI-acLDL uptake (MFI)
0 20 40 60 80 100 120
DTF NOB
+ 30 nM PMA
**
**
**
**
(Figure 5AB)
a monocyte
b vehicle
c DTF 10 μM
d DTF 20 μM
e NOB 10 μM
f NOB 20 μM
C
D
concentration (μM)
monocytes vehicle 10 20
DiI-oxLDL uptake (MFI)
0 100 200 300 400
DTF NOB
+ PMA (30 nM)
**
**
**
**
*
(Figure 5CD)
a monocyte
b vehicle
c DTF 10 μM
d DTF 20 μM
e NOB 10 μM
f NOB 20 μM
concentration (μM)
vehicle 10 20
CD36 mRNA expression (fold of macrophages)
0.0
SR-A mRNA expression (fold of macrophages)
0.0
C
(Figure 6 C)
a blank control b vehicle
c DTF 10 μM d DTF 20 μM
e NOB 10 μM f NOB 20 μM
[1] P. Libby, Inflammation in atherosclerosis, Nature 420 (2002) 868-874.
[2] M. Kaplan, M. Aviram, Oxidized low density lipoprotein: atherogenic and proinflammatory characteristics during macrophage foam cell formation. An inhibitory role for nutritional antioxidants and serum paraoxonase, Clin. Chem.
Lab. Med. 37 (1999) 777-787.
[3] R. Ross, Atherosclerosis--an inflammatory disease, N. Engl. J. Med. 340 (1999) 115-126.
[4] Y. Yamada, T. Doi, T. Hamakubo, T. Kodama, Scavenger receptor family proteins: roles for atherosclerosis, host defence and disorders of the central nervous system, Cell. Mol. Life Sci. 54 (1998) 628-640.
[5] W.J. de Villiers, E.J. Smart, Macrophage scavenger receptors and foam cell formation, J. Leukoc. Biol. 66 (1999) 740-746.
[6] G. Endemann, L.W. Stanton, K.S. Madden, C.M. Bryant, R.T. White, A.A.
Protter, CD36 is a receptor for oxidized low density lipoprotein, J Biol Chem 268 (1993) 11811-11816.
[7] Y. Zeng, N. Tao, K.N. Chung, J.E. Heuser, D.M. Lublin, Endocytosis of oxidized low density lipoprotein through scavenger receptor CD36 utilizes a lipid raft pathway that does not require caveolin-1, J. Biol. Chem. 278 (2003) 45931-45936.
[8] A. Nakata, Y. Nakagawa, M. Nishida, S. Nozaki, J. Miyagawa, T. Nakagawa, R. Tamura, K. Matsumoto, K. Kameda-Takemura, S. Yamashita, Y.
Matsuzawa, CD36, a novel receptor for oxidized low-density lipoproteins, is highly expressed on lipid-laden macrophages in human atherosclerotic aorta, Arterioscler Thromb Vasc Biol 19 (1999) 1333-1339.
[9] D.P. Hajjar, M.E. Haberland, Lipoprotein trafficking in vascular cells.
Molecular Trojan horses and cellular saboteurs, J. Biol. Chem. 272 (1997) 22975-22978.
[10] J. Han, D.P. Hajjar, M. Febbraio, A.C. Nicholson, Native and modified low density lipoproteins increase the functional expression of the macrophage class B scavenger receptor, CD36, J. Biol. Chem. 272 (1997) 21654-21659.
[11] N. Platt, S. Gordon, Is the class A macrophage scavenger receptor (SR-A) multifunctional? - The mouse's tale, J. Clin. Invest. 108 (2001) 649-654.
[12] N. Kume, T. Murase, H. Moriwaki, T. Aoyama, T. Sawamura, T. Masaki, T.
Kita, Inducible expression of lectin-like oxidized LDL receptor-1 in vascular endothelial cells, Circ. Res. 83 (1998) 322-327.
[13] H. Kataoka, N. Kume, S. Miyamoto, M. Minami, H. Moriwaki, T. Murase, T.
Sawamura, T. Masaki, N. Hashimoto, T. Kita, Expression of lectinlike
oxidized low-density lipoprotein receptor-1 in human atherosclerotic lesions, Circulation 99 (1999) 3110-3117.
[14] H. Moriwaki, N. Kume, T. Sawamura, T. Aoyama, H. Hoshikawa, H. Ochi, E.
Nishi, T. Masaki, T. Kita, Ligand specificity of LOX-1, a novel endothelial receptor for oxidized low density lipoprotein, Arterioscler. Thromb. Vasc.
Biol. 18 (1998) 1541-1547.
[15] M. Febbraio, E.A. Podrez, J.D. Smith, D.P. Hajjar, S.L. Hazen, H.F. Hoff, K.
Sharma, R.L. Silverstein, Targeted disruption of the class B scavenger
receptor CD36 protects against atherosclerotic lesion development in mice, J.
Clin. Invest. 105 (2000) 1049-1056.
[16] H. Suzuki, Y. Kurihara, M. Takeya, N. Kamada, M. Kataoka, K. Jishage, O.
Ueda, H. Sakaguchi, T. Higashi, T. Suzuki, Y. Takashima, Y. Kawabe, O.
Cynshi, Y. Wada, M. Honda, H. Kurihara, H. Aburatani, T. Doi, A.
Matsumoto, S. Azuma, T. Noda, Y. Toyoda, H. Itakura, Y. Yazaki, T.
Kodama, et al., A role for macrophage scavenger receptors in atherosclerosis and susceptibility to infection, Nature 386 (1997) 292-296.
[17] V.R. Babaev, L.A. Gleaves, K.J. Carter, H. Suzuki, T. Kodama, S. Fazio, M.F.
Linton, Reduced atherosclerotic lesions in mice deficient for total or macrophage-specific expression of scavenger receptor-A, Arterioscler.
Thromb. Vasc. Biol. 20 (2000) 2593-2599.
[18] H. Sakaguchi, M. Takeya, H. Suzuki, H. Hakamata, T. Kodama, S. Horiuchi, S. Gordon, L.J. van der Laan, G. Kraal, S. Ishibashi, N. Kitamura, K.
Takahashi, Role of macrophage scavenger receptors in diet-induced atherosclerosis in mice, Lab. Invest. 78 (1998) 423-434.
[19] J.L. Mehta, N. Sanada, C.P. Hu, J. Chen, A. Dandapat, F. Sugawara, H. Satoh, K. Inoue, Y. Kawase, K. Jishage, H. Suzuki, M. Takeya, L. Schnackenberg, R.
Beger, P.L. Hermonat, M. Thomas, T. Sawamura, Deletion of LOX-1 reduces atherogenesis in LDLR knockout mice fed high cholesterol diet, Circ. Res.
100 (2007) 1634-1642.
[20] C. Hu, J. Chen, A. Dandapat, Y. Fujita, N. Inoue, Y. Kawase, K. Jishage, H.
Suzuki, D. Li, P.L. Hermonat, T. Sawamura, J.L. Mehta, LOX-1 abrogation reduces myocardial ischemia-reperfusion injury in mice, J. Mol. Cell. Cardiol.
44 (2008) 76-83.
[21] M. Aviram, Macrophage foam cell formation during early atherogenesis is determined by the balance between pro-oxidants and anti-oxidants in arterial cells and blood lipoproteins, Antioxid. Redox. Signal. 1 (1999) 585-594.
[22] M.N. Diaz, B. Frei, J.A. Vita, J.F. Keaney, Jr., Antioxidants and atherosclerotic heart disease, N. Engl. J. Med. 337 (1997) 408-416.
[23] B. Fuhrman, N. Volkova, R. Coleman, M. Aviram, Grape Powder Polyphenols Attenuate Atherosclerosis Development in Apolipoprotein E Deficient (E0) Mice and Reduce Macrophage Atherogenicity, J. Nutr. 135 (2005) 722-728.
[24] B. Fuhrman, M. Aviram, Anti-atherogenicity of nutritional antioxidants, IDrugs 4 (2001) 82-92.
[25] B. Fuhrman, M. Aviram, Flavonoids protect LDL from oxidation and attenuate atherosclerosis, Curr. Opin. Lipidol. 12 (2001) 41-48.
[26] A. Munteanu, J.M. Zingg, R. Ricciarelli, A. Azzi, CD36 overexpression in ritonavir-treated THP-1 cells is reversed by alpha-tocopherol, Free Radic. Biol.
Med. 38 (2005) 1047-1056.
[27] Y. Kawai, T. Nishikawa, Y. Shiba, S. Saito, K. Murota, N. Shibata, M.
Kobayashi, M. Kanayama, K. Uchida, J. Terao, Macrophage as a target of quercetin glucuronides in human atherosclerotic arteries: implication in the anti-atherosclerotic mechanism of dietary flavonoids, J. Biol. Chem. 283 (2008) 9424-9434.
[28] T.W. Lian, L. Wang, Y.H. Lo, I.J. Huang, M.J. Wu, Fisetin, morin and myricetin attenuate CD36 expression and oxLDL uptake in U937-derived macrophages, Biochim. Biophys. Acta 1781 (2008) 601-609.
[29] A. Munteanu, M. Taddei, I. Tamburini, E. Bergamini, A. Azzi, J.M. Zingg, Antagonistic effects of oxidized low density lipoprotein and alpha-tocopherol on CD36 scavenger receptor expression in monocytes: involvement of protein kinase B and peroxisome proliferator-activated receptor-gamma, J. Biol.
Chem. 281 (2006) 6489-6497.
[30] S. Devaraj, I. Hugou, I. Jialal, Alpha-tocopherol decreases CD36 expression in human monocyte-derived macrophages, J. Lipid Res. 42 (2001) 521-527.
[31] E.M. Kurowska, J.A. Manthey, Hypolipidemic effects and absorption of citrus polymethoxylated flavones in hamsters with diet-induced
hypercholesterolemia, J. Agric. Food Chem. 52 (2004) 2879-2886.
[32] J.M. Roza, Z. Xian-Liu, N. Guthrie, Effect of citrus flavonoids and tocotrienols on serum cholesterol levels in hypercholesterolemic subjects, Altern. Ther. Health Med. 13 (2007) 44-48.
[33] L.J. Wilcox, N.M. Borradaile, L.E. de Dreu, M.W. Huff, Secretion of
hepatocyte apoB is inhibited by the flavonoids, naringenin and hesperetin, via reduced activity and expression of ACAT2 and MTP, J. Lipid Res. 42 (2001) 725-734.
[34] B. Morin, L.A. Nichols, K.M. Zalasky, J.W. Davis, J.A. Manthey, L.J.
Holland, The Citrus Flavonoids Hesperetin and Nobiletin Differentially Regulate Low Density Lipoprotein Receptor Gene Transcription in HepG2
Liver Cells, J. Nutr. 138 (2008) 1274-1281.
[35] A. Eguchi, A. Murakami, H. Ohigashi, Nobiletin, a citrus flavonoid,
suppresses phorbol ester-induced expression of multiple scavenger receptor genes in THP-1 human monocytic cells, FEBS Lett. 580 (2006) 3321-3328.
[36] S.C. Whitman, E.M. Kurowska, J.A. Manthey, A. Daugherty, Nobiletin, a citrus flavonoid isolated from tangerines, selectively inhibits class A scavenger receptor-mediated metabolism of acetylated LDL by mouse macrophages, Atherosclerosis 178 (2005) 25-32.
[37] C.S. Lai, S. Li, C.Y. Chai, C.Y. Lo, S. Dushenkov, C.T. Ho, M.H. Pan, Y.J.
Wang, Anti-inflammatory and antitumor promotional effects of a novel urinary metabolite, 3',4'-didemethylnobiletin, derived from nobiletin, Carcinogenesis 29 (2008) 2415-2424.
[38] S. Li, S. Sang, M.H. Pan, C.S. Lai, C.Y. Lo, C.S. Yang, C.T. Ho,
Anti-inflammatory property of the urinary metabolites of nobiletin in mouse, Bioorg. Med. Chem. Lett. 17 (2007) 5177-5181.
[39] A. Eguchi, A. Murakami, S. Li, C.T. Ho, H. Ohigashi, Suppressive effects of demethylated metabolites of nobiletin on phorbol ester-induced expression of scavenger receptor genes in THP-1 human monocytic cells, Biofactors 31 (2007) 107-116.
[40] H. Esterbauer, G. Striegl, H. Puhl, M. Rotheneder, Continuous monitoring of in vitro oxidation of human low density lipoprotein, Free Radic. Res. Commun.
6 (1989) 67-75.
[41] G. Knipping, M. Rotheneder, G. Striegl, H. Esterbauer, Antioxidants and resistance against oxidation of porcine LDL subfractions, J. Lipid Res. 31 (1990) 1965-1972.
[42] L. Lanningham-Foster, C. Chen, D.S. Chance, G. Loo, Grape extract inhibits lipid peroxidation of human low density lipoprotein, Biol. Pharm. Bull. 18 (1995) 1347-1351.
[43] A.M. Gotto, Jr., H.J. Pownall, R.J. Havel, Introduction to the plasma lipoproteins, Methods Enzymol 128 (1986) 3-41.
[44] M.M. Bradford, A rapid and sensitive method for the quantitation of
microgram quantities of protein utilizing the principle of protein-dye binding, Anal Biochem 72 (1976) 248-254.
[45] H.A. Kleinveld, H.L. Hak-Lemmers, A.F. Stalenhoef, P.N. Demacker, Improved measurement of low-density-lipoprotein susceptibility to copper-induced oxidation: application of a short procedure for isolating low-density lipoprotein, Clin Chem 38 (1992) 2066-2072.
[46] M. Hermann, B. Gmeiner, Altered susceptibility to in vitro oxidation of LDL
in LDL complexes and LDL aggregates, Ann N Y Acad Sci 683 (1993) 363-364.
[47] B. Fuhrman, A. Partoush, N. Volkova, M. Aviram, Ox-LDL induces monocyte-to-macrophage differentiation in vivo: Possible role for the macrophage colony stimulating factor receptor (M-CSF-R), Atherosclerosis 196 (2008) 598-607.
[48] X. Wang, B. Seed, A PCR primer bank for quantitative gene expression analysis, Nucl. Acids Res. 31 (2003) e154.
[49] D. Teupser, J. Thiery, A.K. Walli, D. Seidel, Determination of LDL- and scavenger-receptor activity in adherent and non-adherent cultured cells with a new single-step fluorometric assay, Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism 1303 (1996) 193-198.
[50] S.K. Basu, J.L. Goldstein, G.W. Anderson, M.S. Brown, Degradation of cationized low density lipoprotein and regulation of cholesterol metabolism in homozygous familial hypercholesterolemia fibroblasts, Proc. Natl. Acad. Sci.
U. S. A. 73 (1976) 3178-3182.
[51] U.P. Steinbrecher, H.F. Zhang, M. Lougheed, Role of oxidatively modified LDL in atherosclerosis, Free Radic Biol Med 9 (1990) 155-168.
[52] P.M. Abuja, H. Esterbauer, Simulation of lipid peroxidation in low-density lipoprotein by a basic "skeleton" of reactions, Chem. Res. Toxicol. 8 (1995) 753-763.
[53] O. Ziouzenkova, A. Sevanian, P.M. Abuja, P. Ramos, H. Esterbauer, Copper can promote oxidation of LDL by markedly different mechanisms, Free Radic.
Biol. Med. 24 (1998) 607-623.
[54] J. Auwerx, The human leukemia cell line, THP-1: a multifacetted model for the study of monocyte-macrophage differentiation, Experientia 47 (1991) 22-31.
[55] J. Carmichael, W.G. DeGraff, A.F. Gazdar, J.D. Minna, J.B. Mitchell, Evaluation of a tetrazolium-based semiautomated colorimetric assay:
assessment of chemosensitivity testing, Cancer Res. 47 (1987) 936-942.
[56] M.A. Arnaout, Structure and function of the leukocyte adhesion molecules CD11/CD18, Blood 75 (1990) 1037-1050.
[57] V.V. Kunjathoor, M. Febbraio, E.A. Podrez, K.J. Moore, L. Andersson, S.
Koehn, J.S. Rhee, R. Silverstein, H.F. Hoff, M.W. Freeman, Scavenger receptors class A-I/II and CD36 are the principal receptors responsible for the uptake of modified low density lipoprotein leading to lipid loading in
macrophages, J. Biol. Chem. 277 (2002) 49982-49988.
[58] M.T. Quinn, S. Parthasarathy, L.G. Fong, D. Steinberg, Oxidatively modified
low density lipoproteins: a potential role in recruitment and retention of monocyte/macrophages during atherogenesis, Proc. Natl. Acad. Sci. U. S. A.
84 (1987) 2995-2998.
[59] J.M. Hayden, L. Brachova, K. Higgins, L. Obermiller, A. Sevanian, S.
Khandrika, P.D. Reaven, Induction of monocyte differentiation and foam cell formation in vitro by 7-ketocholesterol, J. Lipid Res. 43 (2002) 26-35.
[60] J. Frostegard, J. Nilsson, A. Haegerstrand, A. Hamsten, H. Wigzell, M.
Gidlund, Oxidized low density lipoprotein induces differentiation and adhesion of human monocytes and the monocytic cell line U937, Proc. Natl.
Acad. Sci. U. S. A. 87 (1990) 904-908.
[61] J. Pou, A. Rebollo, N. Roglans, R.M. Sanchez, M. Vazquez-Carrera, J.C.
Laguna, J. Pedro-Botet, M. Alegret, Ritonavir increases CD36, ABCA1 and CYP27 expression in THP-1 macrophages, Exp Biol Med (Maywood) 233 (2008) 1572-1582.
[62] Y.C. Tu, T.W. Lian, J.H. Yen, Z.T. Chen, M.J. Wu, Antiatherogenic effects of kaempferol and rhamnocitrin, J. Agric. Food Chem. 55 (2007) 9969-9976.
[63] M. Vinals, I. Bermudez, G. Llaverias, M. Alegret, R.M. Sanchez, M.
Vazquez-Carrera, J.C. Laguna, Aspirin increases CD36, SR-BI, and ABCA1 expression in human THP-1 macrophages, Cardiovasc. Res. 66 (2005) 141-149.
[64] D. Steinberg, S. Parthasarathy, T. Carew, J. Khoo, J. Witztum, Beyond cholesterol. Modifications of lowdensity lipoprotein that increase its atherogenicity, N. Engl. J. Med. 320 (1989) 915-924.
[65] C.V. de Whalley, S.M. Rankin, J.R. Hoult, W. Jessup, D.S. Leake, Flavonoids inhibit the oxidative modification of low density lipoproteins by macrophages, Biochem Pharmacol 39 (1990) 1743-1750.
[66] M.R. Safari, N. Sheikh, Effects of flavonoids on the susceptibility of low-density lipoprotein to oxidative modification, Prostaglandins Leukot Essent Fatty Acids 69 (2003) 73-77.
[67] G.A. Naderi, S. Asgary, N. Sarraf-Zadegan, H. Shirvany, Anti-oxidant effect of flavonoids on the susceptibility of LDL oxidation, Mol Cell Biochem 246 (2003) 193-196.
[68] S. Miura, J. Watanabe, M. Sano, T. Tomita, T. Osawa, Y. Hara, I. Tomita, Effects of various natural antioxidants on the Cu(2+)-mediated oxidative modification of low density lipoprotein, Biol Pharm Bull 18 (1995) 1-4.
[69] U.P. Steinbrecher, J.L. Witztum, S. Parthasarathy, D. Steinberg, Decrease in reactive amino groups during oxidation or endothelial cell modification of LDL. Correlation with changes in receptor-mediated catabolism,
Arteriosclerosis 7 (1987) 135-143.
[70] D. Steinberg, Low density lipoprotein oxidation and its pathobiological significance, J. Biol. Chem. 272 (1997) 20963-20966.
[71] H. Esterbauer, Cytotoxicity and genotoxicity of lipid-oxidation products, Am.
J. Clin. Nutr. 57 (1993) 779S-785S; discussion 785S-786S.
[72] T.T. Tuomisto, M.S. Riekkinen, H. Viita, A.-L. Levonen, S. Yl?Herttuala, Analysis of gene and protein expression during monocyte-macrophage differentiation and cholesterol loading--cDNA and protein array study, Atherosclerosis 180 (2005) 283-291.
[73] J. Zhang, W. Chu, I. Crandall, Lipoprotein binding preference of CD36 is altered by filipin treatment, Lipids Health Dis. 7 (2008) 23.
[74] D. Klein, Quantification using real-time PCR technology: applications and limitations, Trends Mol. Med. 8 (2002) 257-260.
[75] A. Munteanu, M. Taddei, I. Tamburini, E. Bergamini, A. Azzi, J.-M. Zingg, Antagonistic effects of oxLDL and alpha -tocopherol on CD36 scavenger receptor expression in monocytes; involvement of PKB and PPARgamma, J.
Biol. Chem. (2006) M508799200.
[76] H. Schwende, E. Fitzke, P. Ambs, P. Dieter, Differences in the state of differentiation of THP-1 cells induced by phorbol ester and
1,25-dihydroxyvitamin D3, J. Leukoc. Biol. 59 (1996) 555-561.
[77] N. Koga, M. Matsuo, C. Ohta, K. Haraguchi, M. Matsuoka, Y. Kato, T. Ishii, M. Yano, H. Ohta, Comparative Study on Nobiletin Metabolism with Liver Microsomes from Rats, Guinea Pigs and Hamsters and Rat Cytochrome P450, Biol. Pharm. Bull. 30 (2007) 2317-2323.