Abstract
Purpose
Atherosclerosis is associated with reduced mononuclear cell (MNC) telomere length, and senescent cells have been detected in atherosclerotic plaques. Rice bran is a source of γ-oryzanol, phytosterols and tocols with potential lipid-lowering, antioxidant and anti-inflammatory activities. Here, we tested the hypothesis that rice bran enzymatic extract (RBEE) impacts on apoptosis, telomere length and atherogenesis in mice.
Methods
Seven-week-old male ApoE−/− mice were fed high-fat diet (HFD) or isocaloric HFD supplemented with 5 % (w/w) RBEE for 23 weeks. Wild-type mice of the same age were kept under standard diet as controls.
Results
RBEE treatment reduced total cholesterol (19.24 ± 1.63 vs 24.49 ± 1.71 mmol/L) and triglycerides (1.13 ± 0.18 vs 1.75 ± 0.22 mmol/L) and augmented HDL-cholesterol (1.86 ± 0.20 vs 1.07 ± 0.20 mmol/L). RBEE attenuated macrophage infiltration by 56.69 ± 4.65 % and plaque development (7737 ± 836 vs 12,040 ± 1001 μm2) in the aortic sinus. In the aorta, RBEE treatment reduced expression of the apoptosis pathway components p16, p53 and bax/bcl-2 ratio. RBEE prevented apoptosis of aortic endothelial cells (2.81 ± 0.71–1.14 ± 0.35 apoptotic nuclei/ring for ApoE−/− HFD and ApoE−/− HFD 5 % RBEE, respectively). In contrast, MNC of RBEE-fed mice exhibited enhanced apoptosis marker expression with increased p53 and bax/bcl-2 protein levels. Compared to WT, ApoE−/− mice on HFD were characterized by significant telomere shortening in aorta (11 ± 2 %) and MNC (73 ± 7 %), which was reduced by supplementation with RBEE (aorta: 40 ± 7 %; MNC: 105 ± 10 %). Expression of telomere repeat-binding factor 2 was increased in RBEE-fed mice.
Conclusion
Long-term food supplementation with RBEE lowers cholesterol and prevents atherosclerotic plaque development in ApoE−/− mice. Differential regulation of vascular and MNC apoptosis and senescence were identified as potential mechanisms.
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References
Minamino T, Komuro I (2007) Vascular cell senescence: contribution to atherosclerosis. Circ Res 100(1):15–26
Lopez-Otin C, Blasco MA, Partridge L, Serrano M, Kroemer G (2013) The hallmarks of aging. Cell 153(6):1194–1217
Fyhrquist F, Saijonmaa O, Strandberg T (2013) The roles of senescence and telomere shortening in cardiovascular disease. Nat Rev Cardiol 10(5):274–283
Aviv A, Levy D (2012) Telomeres, atherosclerosis, and the hemothelium: the longer view. Annu Rev Med 63:293–301
Werner C, Fürster T, Widmann T, Pöss J, Roggia C, Hanhoun M, Scharhag J, Büchner N, Meyer T, Kindermann W et al (2009) Physical exercise prevents cellular senescence in circulating leukocytes and in the vessel wall. Circulation 120(24):2438–2447
Werner C, Gensch C, Pöss J, Haendeler J, Böhm M, Laufs U (2011) Pioglitazone activates aortic telomerase and prevents stress-induced endothelial apoptosis. Atherosclerosis 216(1):23–34
Chen S, Lin J, Matsuguchi T, Blackburn E, Yeh F, Best LG, Devereux RB, Lee ET, Howard BV, Roman MJ et al (2014) Short leukocyte telomere length predicts incidence and progression of carotid atherosclerosis in American Indians: the strong heart family study. Aging (Albany NY) 6(5):414–427
Huzen J, Peeters W, de Boer RA, Moll FL, Wong LS, Codd V, de Kleijn DP, de Smet BJ, van Veldhuisen DJ, Samani NJ et al (2011) Circulating leukocyte and carotid atherosclerotic plaque telomere length: interrelation, association with plaque characteristics, and restenosis after endarterectomy. Arterioscler Thromb Vasc Biol 31(5):1219–1225
Brouilette SW, Moore JS, McMahon AD, Thompson JR, Ford I, Shepherd J, Packard CJ, Samani NJ (2007) Telomere length, risk of coronary heart disease, and statin treatment in the West of Scotland Primary Prevention Study: a nested case-control study. Lancet 369(9556):107–114
O’Donnell CJ, Demissie S, Kimura M, Levy D, Gardner JP, White C, D’Agostino RB, Wolf PA, Polak J, Cupples LA et al (2008) Leukocyte telomere length and carotid artery intimal medial thickness: the Framingham Heart Study. Arterioscler Thromb Vasc Biol 28(6):1165–1171
Willeit P, Willeit J, Brandstatter A, Ehrlenbach S, Mayr A, Gasperi A, Weger S, Oberhollenzer F, Reindl M, Kronenberg F et al (2010) Cellular aging reflected by leukocyte telomere length predicts advanced atherosclerosis and cardiovascular disease risk. Arterioscler Thromb Vasc Biol 30(8):1649–1656
Baragetti A, Palmen J, Garlaschelli K, Grigore L, Pellegatta F, Tragni E, Catapano AL, Humphries SE, Norata GD, Talmud PJ (2015) Telomere shortening over 6 years is associated with increased subclinical carotid vascular damage and worse cardiovascular prognosis in the general population. J Intern Med 277(4):478–487
Butt HZ, Atturu G, London NJ, Sayers RD, Bown MJ (2010) Telomere length dynamics in vascular disease: a review. Eur J Vasc Endovasc Surg 40(1):17–26
Poch E, Carbonell P, Franco S, Diez-Juan A, Blasco MA, Andres V (2004) Short telomeres protect from diet-induced atherosclerosis in apolipoprotein E-null mice. FASEB J 18(2):418–420
Soory M (2012) Nutritional antioxidants and their applications in cardiometabolic diseases. Infect Disord Drug Targets 12(5):388–401
Riccioni G, Speranza L, Pesce M, Cusenza S, D’Orazio N, Glade MJ (2012) Novel phytonutrient contributors to antioxidant protection against cardiovascular disease. Nutrition 28(6):605–610
Chen G, Wang H, Zhang X, Yang ST (2014) Nutraceuticals and functional foods in the management of hyperlipidemia. Crit Rev Food Sci Nutr 54(9):1180–1201
Goufo P, Trindade H (2014) Rice antioxidants: phenolic acids, flavonoids, anthocyanins, proanthocyanidins, tocopherols, tocotrienols, gamma-oryzanol, and phytic acid. Food Sci Nutr 2(2):75–104
Rukmini C, Raghuram TC (1991) Nutritional and biochemical aspects of the hypolipidemic action of rice bran oil: a review. J Am Coll Nutr 10(6):593–601
Islam MS, Nagasaka R, Ohara K, Hosoya T, Ozaki H, Ushio H, Hori M (2011) Biological abilities of rice bran-derived antioxidant phytochemicals for medical therapy. Curr Top Med Chem 11(14):1847–1853
Friedman M (2013) Rice brans, rice bran oils, and rice hulls: composition, food and industrial uses, and bioactivities in humans, animals, and cells. J Agric Food Chem 61(45):10626–10641
Parrado J, Miramontes E, Jover M, Gutierrez JF, de Teran L, Bautista J (2006) Preparation of a rice bran enzymatic extract with potential use as functional food. Food Chem 98(4):742–748
Cawthon RM, Smith KR, O’Brien E, Sivatchenko A, Kerber RA (2003) Association between telomere length in blood and mortality in people aged 60 years or older. Lancet 361(9355):393–395
Kim NW, Wu F (1997) Advances in quantification and characterization of telomerase activity by the telomeric repeat amplification protocol (TRAP). Nucleic Acids Res 25(13):2595–2597
Werner C, Hanhoun M, Widmann T, Kazakov A, Semenov A, Pöss J, Bauersachs J, Thum T, Pfreundschuh M, Müller P et al (2008) Effects of physical exercise on myocardial telomere-regulating proteins, survival pathways, and apoptosis. J Am Coll Cardiol 52(6):470–482
Martinet W, Kockx MM (2001) Apoptosis in atherosclerosis: focus on oxidized lipids and inflammation. Curr Opin Lipidol 12(5):535–541
Revilla E, Santa-María C, Miramontes E, Bautista Lorite J, García-Martínez A, Cremades O, Cert R, Parrado J (2009) Nutraceutical composition, antioxidant activity and hypocholesterolemic effect of a water-soluble enzymatic extract from rice bran. Food Res Int 42:387–393
Justo ML, Rodriguez-Rodriguez R, Claro CM, Alvarez de Sotomayor M, Parrado J, Herrera MD (2013) Water-soluble rice bran enzymatic extract attenuates dyslipidemia, hypertension and insulin resistance in obese Zucker rats. Eur J Nutr 52(2):789–797
Justo ML, Candiracci M, Dantas AP, de Sotomayor MA, Parrado J, Vila E, Herrera MD, Rodriguez-Rodriguez R (2013) Rice bran enzymatic extract restores endothelial function and vascular contractility in obese rats by reducing vascular inflammation and oxidative stress. J Nutr Biochem 24(8):1453–1461
Justo ML, Claro C, Vila E, Herrera MD, Rodriguez-Rodriguez R (2014) Microvascular disorders in obese Zucker rats are restored by a rice bran diet. Nutr Metab Cardiovasc Dis 24(5):524–531
Candiracci M, Justo ML, Castano A, Rodriguez-Rodriguez R, Herrera MD (2014) Rice bran enzymatic extract-supplemented diets modulate adipose tissue inflammation markers in Zucker rats. Nutrition 30(4):466–472
Ausman LM, Rong N, Nicolosi RJ (2005) Hypocholesterolemic effect of physically refined rice bran oil: studies of cholesterol metabolism and early atherosclerosis in hypercholesterolemic hamsters. J Nutr Biochem 16(9):521–529
Wang YX, Li Y, Sun AM, Wang FJ, Yu GP (2014) Hypolipidemic and antioxidative effects of aqueous enzymatic extract from rice bran in rats fed a high-fat and-cholesterol diet. Nutrients 6(9):3696–3710
Berger A, Rein D, Schafer A, Monnard I, Gremaud G, Lambelet P, Bertoli C (2005) Similar cholesterol-lowering properties of rice bran oil, with varied gamma-oryzanol, in mildly hypercholesterolemic men. Eur J Nutr 44(3):163–173
Most MM, Tulley R, Morales S, Lefevre M (2005) Rice bran oil, not fiber, lowers cholesterol in humans. Am J Clin Nutr 81(1):64–68
Chen CW, Cheng HH (2006) A rice bran oil diet increases LDL-receptor and HMG-CoA reductase mRNA expressions and insulin sensitivity in rats with streptozotocin/nicotinamide-induced type 2 diabetes. J Nutr 136(6):1472–1476
Lakatta EG, Levy D (2003) Arterial and cardiac aging: major shareholders in cardiovascular disease enterprises: part I: aging arteries: a “set up” for vascular disease. Circulation 107(1):139–146
Zhang W, Hui R, Yang S (2014) Telomeres, cardiovascular aging, and potential intervention for cellular senescence. Sci China Life Sci 57(8):858–862
Nakagawa K, Eitsuka T, Inokuchi H, Miyazawa T (2004) DNA chip analysis of comprehensive food function: inhibition of angiogenesis and telomerase activity with unsaturated vitamin E, tocotrienol. BioFactors 21(1–4):5–10
Eitsuka T, Nakagawa K, Miyazawa T (2006) Down-regulation of telomerase activity in DLD-1 human colorectal adenocarcinoma cells by tocotrienol. Biochem Biophys Res Commun 348(1):170–175
Gizard F, Heywood EB, Findeisen HM, Zhao Y, Jones KL, Cudejko C, Post GR, Staels B, Bruemmer D (2011) Telomerase activation in atherosclerosis and induction of telomerase reverse transcriptase expression by inflammatory stimuli in macrophages. Arterioscler Thromb Vasc Biol 31(2):245–252
von Zglinicki T (2002) Oxidative stress shortens telomeres. Trends Biochem Sci 27(7):339–344
Parrado J, Miramontes E, Jover M, Marquez JC, Angeles Mejias M, Collantes De Teran L, Absi E, Bautista J (2003) Prevention of brain protein and lipid oxidation elicited by a water-soluble oryzanol enzymatic extract derived from rice bran. Eur J Nutr 42(6):307–314
Santa-Maria C, Revilla E, Miramontes E, Bautista J, Garcia-Martinez A, Romero E, Carballo M, Parrado J (2010) Protection against free radicals (UVB irradiation) of a water-soluble enzymatic extract from rice bran. Study using human keratinocyte monolayer and reconstructed human epidermis. Food Chem Toxicol 48(1):83–88
Brown AL, Zhu X, Rong S, Shewale S, Seo J, Boudyguina E, Gebre AK, Alexander-Miller MA, Parks JS (2012) Omega-3 fatty acids ameliorate atherosclerosis by favorably altering monocyte subsets and limiting monocyte recruitment to aortic lesions. Arterioscler Thromb Vasc Biol 32(9):2122–2130
Dutta P, Courties G, Wei Y, Leuschner F, Gorbatov R, Robbins CS, Iwamoto Y, Thompson B, Carlson AL, Heidt T et al (2012) Myocardial infarction accelerates atherosclerosis. Nature 487(7407):325–329
Henderson AJ, Ollila CA, Kumar A, Borresen EC, Raina K, Agarwal R, Ryan EP (2012) Chemopreventive properties of dietary rice bran: current status and future prospects. Adv Nutr 3(5):643–653
de Nigris F, Franconi F, Maida I, Palumbo G, Anania V, Napoli C (2000) Modulation by alpha- and gamma-tocopherol and oxidized low-density lipoprotein of apoptotic signaling in human coronary smooth muscle cells. Biochem Pharmacol 59(11):1477–1487
Khanduja KL, Avti PK, Kumar S, Mittal N, Sohi KK, Pathak CM (2006) Anti-apoptotic activity of caffeic acid, ellagic acid and ferulic acid in normal human peripheral blood mononuclear cells: a Bcl-2 independent mechanism. Biochim Biophys Acta 2:283–289
Acknowledgments
The authors thank Dr. Juan Parrado (University of Seville) for supplying the rice bran extract (RBEE) and Jennifer Kieffer and Ellen Becker for excellent technical assistance. The study was funded by the Corona foundation via the Stifterverband für die Deutsche Wissenschaft and the Spanish Government. Cristina Perez-Ternero is a recipient of a FPU fellowship from the Spanish Government.
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Approval for all experiments was provided by the Ethic Committee for Animal Experimentation of the University of Seville (Spain) (AGL2013-407791-P). The manuscript does not contain clinical studies or patient data.
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Perez-Ternero, C., Herrera, M.D., Laufs, U. et al. Food supplementation with rice bran enzymatic extract prevents vascular apoptosis and atherogenesis in ApoE−/− mice. Eur J Nutr 56, 225–236 (2017). https://doi.org/10.1007/s00394-015-1074-z
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DOI: https://doi.org/10.1007/s00394-015-1074-z