The aim of the present work was to explore possible protective effects of sulforaphane (SFN) against atherosclerosis development and endothelial dysfunction in hypercholesterolemic rabbits. Rabbits were assigned to three groups of five: group I fed normal chow diet for four weeks, group II fed 1% high cholesterol diet (HCD) and group III fed HCD + SFN (0.25 mg/kg/day). Blood samples were collected for measurement of serum triglycerides (TGs), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), lactate dehydrogenase (LDH) and C-reactive protein (CRP). Aortic malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and total nitrite/nitrate (NOx) were measured. Vascular reactivity and intima/media (I/M) ratio were analyzed. Nuclear factor-kappa B (NF-κB) activation in aortic endothelial cells was identified immunohistochemically. HCD induced significant increases in serum TGs, TC, LDL-C, LDH, and CRP, and aortic MDA and SOD. Moreover, HCD caused significant reductions in serum HDL-C, aortic GSH and NOx. SFN administration significantly decreased HCD-induced elevations in serum TC, LDL-C, CRP, and LDH. while significantly increased HDL-C and GSH levels and normalized aortic SOD and NOx. Additionally, SFN significantly improved rabbit aortic endothelium-dependent relaxation to acetylcholine. Moreover, SFN significantly reduced the elevation in I/M ratio. This effect was confirmed by aortic histopathologic examination. The expression of NF-κB in aortic tissue showed a marked reduction upon treatment with SFN. In conclusion, this study reveals that SFN has the ability to ameliorate HCD-induced atherosclerotic lesions progression and vascular dysfunction, possibly via its lipid-lowering and antioxidant effects and suppression of NF-κB-mediated inflammation.
CastelliWP. Cardiovascular disease in women. Am J Obstet Gynecol1988; 158: 1553–60, 66–7.
2.
WitztumJLSteinbergD. Role of oxidized low density lipoprotein in atherogenesis. J Clin Invest1991; 88: 1785–92.
3.
PrasadKKalraJ. Oxygen free radicals and hypercholesterolemic atherosclerosis: Effect of vitamin E. Am Heart J1993; 125: 958–73.
4.
ManthaSVKalraJPrasadK. Effects of probucol on hypercholesterolemia-induced changes in antioxidant enzymes. Life Sci1996; 58: 503–9.
5.
DuellPB. Prevention of atherosclerosis with dietary antioxidants: fact or fiction?J Nutr1996; 126: 1067S–71S.
6.
KatsikiNManesC. Is there a role for supplemented antioxidants in the prevention of atherosclerosis?Clin Nutr2009; 28: 3–9.
7.
WillcoxBJCurbJDRodriguezBL. Antioxidants in cardiovascular health and disease: key lessons from epidemiologic studies. Am J Cardiol2008; 101: 75D–86D.
8.
RaghavamenonAGarelnabiMBabuSAldrichALitvinovDParthasarathyS. Alpha-tocopherol is ineffective in preventing the decomposition of preformed lipid peroxides and may promote the accumulation of toxic aldehydes: a potential explanation for the failure of antioxidants to affect human atherosclerosis. Antioxid Redox Signal2009; 1.48–1: 1237–1237.
9.
Dinkova-KostovaATTalalayP. Direct and indirect antioxidant properties of inducers of cytoprotective proteins. Mol Nutr Food Res2008; 52: S128–38..
10.
WuLNoyan AshrafMHFacciMWangRPatersonPGFerrieAJuurlinkBH. Dietary approach to attenuate oxidative stress, hypertension, and inflammation in the cardiovascular system. Proc Natl Acad Sci U S A2004; 101: 7094–9.
11.
YochumLKushiLHMeyerKFolsomAR. Dietary flavonoid intake and risk of cardiovascular disease in postmenopausal women. Am J Epidemiol1999; 149: 943–9.
12.
ZhangXShuXOXiangYBYangGLiHGaoJCaiHGaoYTZhengW. Cruciferous vegetable consumption is associated with a reduced risk of total and cardiovascular disease mortality. Am J Clin Nutr2011; 94: 240–6.
13.
XueMQianQAdaikalakoteswariARabbaniNBabaei-JadidiRThornalleyPJ. Activation of NF-E2-related factor-2 reverses biochemical dysfunction of endothelial cells induced by hyperglycemia linked to vascular disease. Diabetes2008; 57: 2809–17.
14.
ThimmulappaRKMaiKHSrisumaSKenslerTWYamamotoMBiswalS. Identification of Nrf2-regulated genes induced by the chemopreventive agent sulforaphane by oligonucleotide microarray. Cancer Res2002; 62: 5196–203.
15.
KeumYS. Regulation of the Keap1/Nrf2 system by chemopreventive sulforaphane: implications of posttranslational modifications. Ann N Y Acad Sci2011; 1229: 184–9.
16.
McMahonMItohKYamamotoMHayesJD. Keap1-dependent proteasomal degradation of transcription factor Nrf2 contributes to the negative regulation of antioxidant response element-driven gene expression. J Biol Chem2003; 278: 21592–600.
17.
AllainCCPoonLSChanCSRichmondWFuPC. Enzymatic determination of total serum cholesterol. Clin Chem1974; 20: 470–5.
18.
FinleyPRSchifmanRBWilliamsRJLichtiDA. Cholesterol in high-density lipoprotein: use of Mg2+/dextran sulfate in its enzymic measurement. Clin Chem1978; 24: 931–3.
19.
FredricksonDSLevyRILeesRS. Fat transport in lipoproteins – an integrated approach to mechanisms and disorders. N Engl J Med1967; 276: 34–42.
20.
FriedewaldWTLevyRIFredricksonDS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem1972; 18: 499–502.
21.
PrasadK. Reduction of serum cholesterol and hypercholesterolemic atherosclerosis in rabbits by secoisolariciresinol diglucoside isolated from flaxseed. Circulation1999; 9: 9.62–1355.
22.
OhkawaHOhishiNYagiK. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem1979; 95: 351–8.
MarklundSL. Superoxide dismutase isoenzymes in tissues and plasma from New Zealand black mice, nude mice and normal BALB/c mice. Mutat Res1985; 148: 129–34.
25.
GrangerDLTaintorRRBoockvarKSHibbsJBJr. Measurement of nitrate and nitrite in biological samples using nitrate reductase and Griess reaction. Meth Enzymol1996; 268: 142–51.
26.
BoriesPNBoriesC. Nitrate determination in biological fluids by an enzymatic one-step assay with nitrate reductase. Clin Chem1995; 41: 904–7.
27.
ChenXLDoddGKunschC. Sulforaphane inhibits TNF-alpha-induced activation of p38 MAP kinase and VCAM-1 and MCP-1 expression in endothelial cells. Inflamm Res2009; 58: 513–21.
28.
KwonJSJoungHKimYSShimYSAhnYJeongMHKeeHJ. Sulforaphane inhibits restenosis by suppressing inflammation and the proliferation of vascular smooth muscle cells. Atherosclerosis2012; 225: 41–9.
29.
PrasadKManthaSVKalraJLeeP. Prevention of hypercholesterolemic atherosclerosis by garlic, an antixoidant. J Cardiovasc Pharmacol Ther1997; 2: 309–20.
30.
AdachiTCohenRA. Decreased aortic glutathione levels may contribute to impaired nitric oxide-induced relaxation in hypercholesterolaemia. Br J Pharmacol2000; 129: 1014–20.
31.
MaXLLopezBLLiuGLChristopherTAGaoFGuoYFeuersteinGZRuffoloRRJr.BaroneFCYueTL. Hypercholesterolemia impairs a detoxification mechanism against peroxynitrite and renders the vascular tissue more susceptible to oxidative injury. Circ Res1997; 80: 894–90.
HennigBChowCK. Lipid peroxidation and endothelial cell injury: implications in atherosclerosis. Free Radic Biol Med1988; 4: 99–106.
35.
Del BoccioGLapennaDPorrecaEPennelliASaviniFFelicianiPRicciGCuccurulloF. Aortic antioxidant defence mechanisms: time-related changes in cholesterol-fed rabbits. Atherosclerosis1990; 81: 127–35.
36.
ManthaSVPrasadMKalraJPrasadK. Antioxidant enzymes in hypercholesterolemia and effects of vitamin E in rabbits. Atherosclerosis1993; 101: 135–44..
37.
ShullSHeintzNHPeriasamyMManoharMJanssenYMMarshJPMossmanBT. Differential regulation of antioxidant enzymes in response to oxidants. J Biol Chem1991; 266: 24398–403.
38.
HassanHM. Biosynthesis and regulation of superoxide dismutases. Free Radic Biol Med1988; 5: 377–85.
39.
KimJYParkHJUmSHSohnEHKimBOMoonEYRheeDKPyoS. Sulforaphane suppresses vascular adhesion molecule-1 expression in TNF-alpha-stimulated mouse vascular smooth muscle cells: involvement of the MAPK, NF-kappaB and AP-1 signaling pathways. Vasc Pharmacol2012; 56: 131–41.
40.
ZhuHJiaZStroblJSEhrichMMisraHPLiY. Potent induction of total cellular and mitochondrial antioxidants and phase 2 enzymes by cruciferous sulforaphane in rat aortic smooth muscle cells: cytoprotection against oxidative and electrophilic stress. Cardiovasc Toxicol2008; 8: 115–25.
41.
FaheyJWTalalayP. Antioxidant functions of sulforaphane: a potent inducer of Phase II detoxication enzymes. Food Chem Toxicol1999; 37: 973–9.
42.
JeonSMParkYBChoiMS. Antihypercholesterolemic property of naringin alters plasma and tissue lipids, cholesterol-regulating enzymes, fecal sterol and tissue morphology in rabbits. Clin Nutr2004; 34–23: 1025–1025.
43.
De La CruzJPQuinteroLVillalobosMASanchez de la CuestaF. Lipid peroxidation and glutathione system in hyperlipemic rabbits: influence of olive oil administration. Biochim Biophys Acta2000; 1485: 36–44.
44.
AikawaMLibbyP. The vulnerable atherosclerotic plaque: pathogenesis and therapeutic approach. Cardiovasc Pathol2004; 13: 125–38.
45.
SinghUJialalI. Oxidative stress and atherosclerosis. Pathophysiology2006; 13: 129–42.
46.
SteinbergDParthasarathySCarewTEKhooJCWitztumJL. Beyond cholesterol. Modifications of low-density lipoprotein that increase its atherogenicity. N Engl J Med1989; 320: 915–24.
47.
CominaciniLPasiniAFGarbinUDavoliATosettiMLCampagnolaMRigoniAPastorinoAMLo CascioVSawamuraT. Oxidized low density lipoprotein (ox-LDL) binding to ox-LDL receptor-1 in endothelial cells induces the activation of NF-kappaB through an increased production of intracellular reactive oxygen species. J Biol Chem2000; 275: 1263–83.
48.
YehMGharaviNMChoiJHsiehXReedEMouillesseauxKPColeALReddySTBerlinerJA. Oxidized phospholipids increase interleukin 8 (IL-8) synthesis by activation of the c-src/signal transducers and activators of transcription (STAT)3 pathway. J Biol Chem2004; 279: 30175–81.
49.
KivelaAMMakinenPIJyrkkanenHKMella-AhoEXiaYKansanenELeinonenHVermaIMYla-HerttualaSLevonenAL. Sulforaphane inhibits endothelial lipase expression through NF-kappaB in endothelial cells. Atherosclerosis2010; 213: 122–8.
50.
SchraderCGraeserACHuebbePWagnerAERimbachG. Allyl isothiocyanate as a potential inducer of paraoxonase-1–studies in cultured hepatocytes and in mice. IUBMB Life2012; 64: 162–8.
51.
WuBJShresthaSOngKLJohnsDDunnLLHouLBarterPJRyeKA. Increasing HDL levels by inhibiting cholesteryl ester transfer protein activity in rabbits with hindlimb ischemia is associated with increased angiogenesis. Int J Cardiol2015; 199: 204–12.
52.
El-AwadyMSSuddekGM. Agmatine ameliorates atherosclerosis progression and endothelial dysfunction in high cholesterol-fed rabbits. J Pharm Pharmacol2014; 66: 835–43.
53.
PasceriVChengJSWillersonJTYehET. Modulation of C-reactive protein-mediated monocyte chemoattractant protein-1 induction in human endothelial cells by anti-atherosclerosis drugs. Circulation2001; 103: 2531–4.
54.
ThomassenMJMeekerDPDeodharSDWiedemannHPBarnaBP. Activation of human monocytes and alveolar macrophages by a synthetic peptide of C-reactive protein. J Immunother Emphasis Tumor Immunol1993; 13: 1–6.
55.
TorzewskiMRistCMortensenRFZwakaTPBienekMWaltenbergerJKoenigWSchmitzGHombachVTorzewskiJ. C-reactive protein in the arterial intima: role of C-reactive protein receptor-dependent monocyte recruitment in atherogenesis. Arterioscler Thromb Vasc Biol2000; 20: 2094–9.
56.
LiuYCHsiehCWWengYCChuangSHHsiehCYWungBS. Sulforaphane inhibition of monocyte adhesion via the suppression of ICAM-1 and NF-kappaB is dependent upon glutathione depletion in endothelial cells. Vasc Pharmacol2008; 48: 54–61.
57.
NallasamyPSiHBabuPVPanDFuYBrookeEAShahHZhenWZhuHLiuDLiYJiaZ. Sulforaphane reduces vascular inflammation in mice and prevents TNF-alpha-induced monocyte adhesion to primary endothelial cells through interfering with the NF-kappaB pathway. J Nutr Biochem2014; 25: 824–33.
58.
LedeburHCParksTP. Transcriptional regulation of the intercellular adhesion molecule-1 gene by inflammatory cytokines in human endothelial cells. Essential roles of a variant NF-kappa B site and p65 homodimers. J Biol Chem1995; 270: 933–43.
59.
BoyleEMJr.KovacichJCCantyTGJr.MorganENChiEVerrierEDPohlmanTH. Inhibition of nuclear factor-kappa B nuclear localization reduces human E-selectin expression and the systemic inflammatory response. Circulation1998; 98: II282–8.
60.
LiYReddyMAMiaoFShanmugamNYeeJKHawkinsDRenBNatarajanR. Role of the histone H3 lysine 4 methyltransferase, SET7/9, in the regulation of NF-kappaB-dependent inflammatory genes. Relevance to diabetes and inflammation. J Biol Chem2008; 283: 26771–81.
61.
WagnerAEBoesch-SaadatmandiCDoseJSchultheissGRimbachG. Anti-inflammatory potential of allyl-isothiocyanate–role of Nrf2, NF-(kappa) B and microRNA-155. J Cell Mol Med2012; 16: 836–43.
62.
HuangCSLinAHLiuCTTsaiCWChangISChenHWLiiCK. Isothiocyanates protect against ox idized LDL-induced endothelial dysfunction by upregulating Nrf2-dependent antioxidation and suppressing NFkappaB activation. Mol Nutr Food Res2013; 57: 1918–30.
63.
JeremyRWMcCarronHSullivanD. Effects of dietary L-arginine on atherosclerosis and endothelium-dependent vasodilatation in the hypercholesterolemic rabbit. Response according to treatment duration, anatomic site, and sex. Circulation1996; 94: 498–506.
64.
ChappellSPLewisMJHendersonAH. Effect of lipid feeding on endothelium dependent relaxation in rabbit aortic preparations. Cardiovasc Res1987; 21: 34–8.
65.
WhiteCRDarley-UsmarVBerringtonWRMcAdamsMGoreJZThompsonJAParksDATarpeyMMFreemanBA. Circulating plasma xanthine oxidase contributes to vascular dysfunction in hypercholesterolemic rabbits. Proc Natl Acad Sci U S A1996; 93: 8745–9.
66.
WilsonSHSimariRDBestPJPetersonTELermanLOAviramMNathKAHolmesDRJr.LermanA. Simvastatin preserves coronary endothelial function in hypercholesterolemia in the absence of lipid lowering. Arterioscler Thromb Vasc Biol2001; 21: 122–8.
ThakurNKHayashiTSumiDKanoHTsunekawaTIguchiA. HMG-CoA reductase inhibitor stabilizes rabbit atheroma by increasing basal NO and decreasing superoxide. Am J Physiol Heart Circ Physiol2001; 281: H75–83.
69.
MillerFJJr.GuttermanDDRiosCDHeistadDDDavidsonBL. Superoxide production in vascular smooth muscle contributes to oxidative stress and impaired relaxation in atherosclerosis. Circ Res1998; 82: 1298–305.
70.
HeissEHSchachnerDWernerERDirschVM. Active NF-E2-related factor (Nrf2) contributes to keep endothelial NO synthase (eNOS) in the coupled state: role of reactive oxygen species (ROS), eNOS, and heme oxygenase (HO-1) levels. J Biol Chem2009; 284: 31579–86.
71.
WangYZhangZSunWTanYLiuYZhengYLiuQCaiLSunJ. Sulforaphane attenuation of type 2 diabetes-induced aortic damage was associated with the upregulation of Nrf2 expression and function. Oxid Med Cell Longev2014; 2014: 123963–123963.
72.
FaheyJWZhangYTalalayP. Broccoli sprouts: an exceptionally rich source of inducers of enzymes that protect against chemical carcinogens. Proc Natl Acad Sci U S A1997; 94: 10367–72.
73.
HanlonNColdhamNGielbertAKuhnertNSauerMJKingLJIoannidesC. Absolute bioavailability and dose-dependent pharmacokinetic behaviour of dietary doses of the chemopreventive isothiocyanate sulforaphane in rat. Br J Nutr2008; 99: 559–64.
74.
YoxallVKentishPColdhamNKuhnertNSauerMJIoannidesC. Modulation of hepatic cytochromes P450 and phase II enzymes by dietary doses of sulforaphane in rats: Implications for its chemopreventive activity. Int J Cancer2005; 117: 356–62.
75.
SongLMorrisonJJBottingNPThornalleyPJ. Analysis of glucosinolates, isothiocyanates, and amine degradation products in vegetable extracts and blood plasma by LC-MS/MS. Anal Biochem2005; 347: 234–43.
