Plasma lipid profile of experimentally induced hyperlipidemic New Zealand white rabbits is not affected by resveratrol.
Presently, cardiovascular diseases are the main causes of morbidity and mortality Morbidity and Mortality can refer to:
pertaining to the muscular tissue of the heart (the myocardium). infarcts and brain vascular accidents. (4) Hyperlipidemia is recognized as an important risk factor to atherosclerosis, and most evidence focuses specifically on hypercholesterolemia. LDL, its main component, is most universally associated to an increased risk of cardiovascular diseases. (4)
Dietary antioxidants have attracted considerable attention as preventive and therapeutic agents. In vivo and in vitro studies using animal models show that the consumption of antioxidants can prevent and delay atherosclerosis progression. Resveratrol res·ver·a·trol
A natural compound found in grapes, mulberries, peanuts, and other plants or food products, especially red wine, that may protect against cancer and cardiovascular disease by acting as an antioxidant, antimutagen, and seems to play an important role in atherosclerosis prevention in human subjects through its inhibitory effect on LDL oxidation, platelet aggregation, and vascular proliferation of smooth muscle cells. (1,6) The hypocholesterolemic action of resveratrol is attributed, at least in part, to an increased excretion of neutral sterols and bile acids into feces. However, there are conflicting results regarding its effects on plasma lipid levels, since some studies have failed to show a reduction in plasma lipid levels induced by such a substance. (20)
The aim of the present study was to evaluate plasma LDL, high-density lipoproteins (HDL), triglycerides, and total cholesterol in experimentally hypercholesterolemic New Zealand white rabbits treated with resveratrol.
MATERIALS AND METHODS
Twenty healthy male adult New Zealand white rabbits whose mean weight was 2.5 kg were used on the study, which was conducted from March 15 to May 15, 2007. The animals were previously fed with regular rabbit ration during one week. Afterwards the animals were divided into four experimental groups containing five animals each, with the following feeding protocol: blank control group (CT) received maintenance diet (a); group R received maintenance diet with simultaneous oral administration of resveratrol (b) (3 mg/kg/day) (a); group CL received 1.5% cholesterol (c) added to the maintenance diet; and group CR received 1.5% cholesterol added to the maintenance diet and simultaneous oral administration of resveratrol (b) (3 mg/kg/ day). For the preparation of hypercholesterolemic diet, a cholesterol powder (b) was added to the previously grounded maintenance diet, mixed, pelleted, and stored at refrigerated units. The dose of resveratrol used was based in previous studies. (19) During the experiment, each animal received daily water ad libitum and 100 g of diet. The experiment was performed under approval of Animal Welfare and Ethics Committee of Universidade Estadual Paulista, Campus de Jaboticabal--UNESP.
Samples of 3 mL of heparinized blood were collected twice monthly from the central ear artery, with the ani mals kept in restraint cages. Those blood samples were used for plasma level determinations of triglycerides, total cholesterol, HDL, and LDL, through enzymatic methods (c), accomplished in Department of Veterinary Clinics and Surgery Laboratory, Campus de Jaboticabal, UNESP HDL levels were measured through selective inhibition technique, while total cholesterol, triglycerides, and LDL through enzymatic / Trinder method. All analytic procedures were made in triplicates. During the experimental period, the animals were weighed on a weekly basis for the adjustments of doses of resveratrol.
Statistical analysis was performed using Statistical Analysis Software (SAS) (d). The results were analyzed through analysis of variance (ANOVA anova
see analysis of variance.
ANOVA Analysis of variance, see there ). A Duncan's test was employed to compare means. A P<0.05 value was considered significant.
Mean values [+ or -] standard deviation of plasma lipid profile obtained in this study are expressed in the Table 1. The values of HDL, LDL, triglycerides, and total cholesterol indicate that the control group was different (P<0.05) from CL and CR groups, but not from R group.
The results obtained in lipid measurements on the first day of the experimental period were similar to all groups. No alterations in plasma lipid profile of the animals fed with regular diet were detected when the values obtained in the beginning and in the end of the experiment were compared, not even in CR group, which showed similar values to CT group in the end of the experiment. Nevertheless, there was a significant increase in total cholesterol, triglycerides, LDL, and HDL in those animals submitted to a hypercholesterolemic diet.
Epidemiological studies show a decreased risk of cardiovascular diseases with the intake of high amounts of polyphenolic substances. Thus, resveratrol contributes with the antioxidant potential and plays an important role in cardiovascular disease prevention in human subjects. The protective effects of resveratrol have been attributed to a number of mechanisms such as antioxidant, anti-inflammatory, anti-proliferative, and anti-thrombotic activities, as well as its hypolipidemic properties. Therefore, considering its antioxidant effects, early studies have proven that polyphenols from red wine were able to reduce plasma oxidation of LDL. (18) A fat-enriched meal prejudices endothelial function for more than four hours, being more severe in coronary disease patients. (22) Such a dysfunction is mainly attributable to an increase in endothelial oxidative stress, which is inhibited by wine antioxidants.
When the in-vitro effects of transresveratrol, some wine-derived phenol compounds, and some antioxidants were studied, a blockage in platelet aggregation and eicosanoid ei·co·sa·noid
Any of the physiologically active substances derived from arachidonic acid, including the prostaglandins, leukotrienes, and thromboxanes. synthesis in human cells was observed, contributing to the cardioprotective effect of resveratrol, mainly against atherosclerosis in coronary disease patients." The beneficial effect of resveratrol on endothelial integrity through an inhibition in VCAM-1 (Vascular Cell Adhesion Molecule 1),ICAM-1 (Intracellular Adhesion Molecule 1), and transcription factor NF- i, (Nuclear Factor Kappa ?) expression, the latter being responsible for pro-inflammatory modulatory mechanisms, is also documented. (5,9,21)
In the present experiment, groups CL and CR shared similarities to all variables studied, although differing from groups CT and R, which were similar. HDL values remained in the species reference range in all groups, only a slight increase being observed in CL and CR groups. LDL, triglycerides, and cholesterol, as expected, reached high levels in hypercholesterolemic groups, but remained in the reference range in CT and R groups, without differing among them.
The analysis of the findings allowed to infer that the use of resveratrol during the experimental period did not affect LDL, HDL, triglycerides, and cholesterol levels, similar to early studies. (17,19,20) Opposite to this evidence, some researchers detected changes in lipid profile induced by different flavonoids flavonoids,
n.pl common plant pigment compounds that act as antioxidants, enhance the effects of vitamin C, and strengthen connective tissue around capillaries. (3,8,10,11,16) and vitamin E (7,15) in various species. Furthermore, a synergic effect between statins and resveratrol was observed in hypercholesterolemic rats, leading to a significative sig·nif·i·ca·tive
1. Tending to signify or indicate; indicative.
2. Having meaning; significant.
sig·nif reduction in total plasma cholesterol, triglycerides, and LDL. (12,14) The use of an analogue of resveratrol (pterostilbene) in hypercholesterolemic hamsters showed a 29% reduction in plasma LDL and a 7% increase in plasma HDL.
In conclusion, resveratrol at 3 mg/kg/day did not show hypolipidemic activity in rabbits experimentally hypercholesterolemic, used as experimental model in atherosclerosis.
Sources of Manufacturers
a. Purina Paulinica, Sao Paulo, Brazil.
b. Vetec Duque de Caxias, Rio de Janeiro, Brazil.
c. LABTEST DIAGNOSTICA S.A, Minas Gerais, Brazil.
d. SAS, Statistics Software v2, version 6.08, Cary, NC.
(1.) Araim O, Ballantyne J, Waterhouse AL, Sumpio BE: 2002, Inhibition of vascular smooth muscle cell proliferation with red wine and red wine polyphenols. J Vasc Surg 35:1226-1232.
(2.) Asciak CR, Hahn S, Diamandis EP, et al.: 1995, The red wine phenolics trans-resveratrol and quercetin quer·ce·tin
A yellow powdered crystalline compound produced synthetically or occurring as a glycoside in the rind and bark of numerous plants, used medicinally to treat abnormal capillary fragility. Also called meletin. block human platelet aggregation and eicosanoid synthesis: implications for protection against coronary heart disease coronary heart disease: see coronary artery disease.
coronary heart disease
or ischemic heart disease
Progressive reduction of blood supply to the heart muscle due to narrowing or blocking of a coronary artery (see atherosclerosis). . Int J Clin Chem 235:207-219.
(3.) Auger C, Teissedre PL, Gerain P, et al.: 2005, Dietary wine phenolics catequin, quercetin, and resveratrol efficiently protect hypercholesterolemic hamsters against aortic fatty streak accumulation. J Agric Food Chem 23:2015-2021.
(4.) Cotran RS, Schoen E. Vasos Sanguineos [Blood Vessels]. In: Cotran RS, Kumar V, Collins T: 2000, Patologia Estrutural e Funcional [Functional and Structural Pathology], 6th ed., pp. 441-460. Guanabara Koogan, Sao Paulo, Brazil.
(5.) Ferrero ME, Bertelli AE, Fulgenzi A, et al.: 1998, Activity in vitro of resveratrol on granulocyte granulocyte /gran·u·lo·cyte/ (gran´u-lo-sit?) granular leukocyte.granulocyt´ic
band-form granulocyte band cell.
n. and monocyte monocyte /mono·cyte/ (mon´o-sit) a mononuclear, phagocytic leukocyte, 13µ to 25µ in diameter, with an ovoid or kidney-shaped nucleus, and azurophilic cytoplasmic granules. adhesion to endotheli um. Am .1 Clin Nutr 68:1208-1214.
(6.) Fremont L: 2000, Minireview: Biological Effects of Resveratrol. Life Sci 66:663-673.
(7.) Hidiroglou N, Gilani GS, Long L, et al.: 2004, The influence of dietary vitamin E, fat, and methionine in blood cholesterol profile, homocysteine levels, and oxidizability of low density lipoprotein Low density lipoprotein (LDL)
A fraction of total serum lipids, the so called "bad" cholesterol.
Mentioned in: Hypercholesterolemia in the gerbil gerbil (jûr`bĭl), small desert rodent found throughout the hot arid regions of Africa and Asia. Also known as sand rats, gerbils have large eyes and powerful, elongated hind limbs upon which they can spring. Gerbils are 3 to 5 in. (7. . J Nutr Biochem 15:730-740.
(8.) Juzwiak S, Wojcicki J, Morkrzycki K, et al.: 2005, Effect of quercetin on experimental hyperlipidemia and atherosclerosis in rabbits. Pharmacol Rep 57:604-609.
(9.) Manna SK, Mukhopadhyay A, Aggarwal BB: 2000, Resveratrol suppresses TNF-induced activation of nuclear transcription factors NF Ib, activator protein-1, and apoptosis: potential role of reactive oxygen intermediates and lipid peroxidation. J Immunol 164:6509-6519.
(10.) Miura D, Miura Y, Yagasaki K: 2003, Hypolipidemic action of dietary resveratrol, a phytoalexin Phytoalexin
Any antibiotic produced by plants in response to microorganisms. Plants use physical and chemical barriers as a first line of defense. When these barriers are breached, however, the plant must actively protect itself by employing a variety of in grapes and red wine, in hepatoma hepatoma /hep·a·to·ma/ (hep?ah-to´mah)
1. a tumor of the liver.
2. hepatocellular carcinoma (malignant h.).
n. pl. bearing rats. Life Sci 73:1393-400.
(11.) Oliveira TT, Nagem TJ, Lopes RM, et al.: 2004, Efeito de diferentes doses de rutina sobre lipideos no soro de coelhos machos e femeas [Effect of different dosis of rutin Ru´tin
n. 1. (Chem.) A glucoside resembling, but distinct from, quercitrin. Rutin is found in the leaves of the rue (Ruta graveolens on serum lipids in male and female rabbits]. Rev Bras Anal Clin 36:213-215.
(12.) Penumathsa SV Tbirunavukkarasu M, Konery S, et al.: 2007, Statin and resveratrol in combination induces cardioprotection against myocardial infarction in hypercholesterolemic rat. J Mot Cell Cardiol 3:508-516.
(13.) Perez G, Pena A, Sala J, et al.: 1998, Acute myocardial infarction acute myocardial infarction (·kyōōtˑ mī·ō·karˑ·dē· case fatality, incidence and mortality rates in a population registry in Gerona, Spain, 1990-1992. Int.1 Epidemiol 27: 599-604.
(14.) Rimando AM, Nagmani R, Feller DR, Yokoyama W: 2005, Pterostilbene, a new agonist for the peroxisome Peroxisome
An intracellular organelle found in all eukaryotes except the archezoa (original lifeforms). In electron micrographs, peroxisomes appear round with a diameter of 0.1–1. proliferator-activated receptor--isoform, lowers plasma lipoproteins and cholesterol in hypercholesterolemic hamsters. Agric Food Chem 53:3403- 3407.
(15.) Schaefer EJ: 2002, Lipoprotein, nutrition, and heart disease. Am J Clin Nutr 75:191-212.
(16.) Shanmuganayagam D, Warner TF, Krueger CG, et al.: 2006, Concord grape juice attenuates platelet aggregation, serum cholesterol and development of atheroma atheroma /ath·er·o·ma/ (ath?er-o´mah) a mass or plaque of degenerated thickened arterial intima, occurring in atherosclerosis.
n. pl. in hypercholesterolemic rabbits. Atherosclerosis 190:135-142.
(17.) Turrens JF, Lariecia J, Nair MG: 1997, Resveratrol has no effect on lipoprotein profile and does not prevent peroxidation of serum lipids in normal rats. Free Radic Res 27:557- 562.
(18.) Wakabayashi Y: 1999, Effect of red wine consumption on low-density lipoprotein oxidation and atherosclerosis in aorta and coronary artery in Watanabe heritable hyperlipidemic rabbits. JAgric Food Chem 46:4724-4730.
(19.) Wang Z, Zou J, Cao K, et al.: 2005, Dealcoholized red wine containing know amounts of resveratrol supresses atheroscle rosis in hypercholesterolemic rabbits without affecting plasma lipid levels. Int J Molec Med 16:533-540.
(20.) Wilson T, Knight TJ, Beitz DC, et al.: 1996, resveratrol promotes atherosclerosis in hypercholesterolemic rabbits. Life Sci 59:115121.
(21.) Wung BS, Hsu MC, Wu CC, Hsieh CW: 2005, Resveratrol suppresses IL-6-induced 1CAM-1 gene expression in endothelial cells: effects on the inhibition of STAT3 phosphorylaton. Life Sci 12:389-397.
(22.) Zhao SP, Liu L, Gao M, et al.: 2001, Impairment of endothelial function after a hight-fat meal in patients with coronary artery disease coronary artery disease, condition that results when the coronary arteries are narrowed or occluded, most commonly by atherosclerotic deposits of fibrous and fatty tissue. . Coron Artery Dis 12:561-565.
Marines de Castro (1)
Angela P M. Veiga
Maria R. Pacheco
From the Sao Paulo State University, Jaboticabal, SP, Brazil (de Castro, Pacheco) and the Federal University of Rio Grande The University of Rio Grande and Rio Grande Community College are twin colleges in Rio Grande, Ohio.
The University of Rio Grande offers a range of courses and majors and is known in the region for its Education and Nursing programs. do Sul, Porto Alegre, RS, Brazil (Veiga).
(1) Corresponding Author: Marines de Castro, Universidade Estadual Paulista (UNESP), Faculty of Agricultural and Veterinary Sciences, Department of Morphology, Via de Acesso Prof Paulo Donato Castellane, s/n, Jaboticabal, Sao Paulo, Brazil 14884-900. Tel: + 55 15 32092654; fax: +55 15 32035150; e-mail: email@example.com.
Table 1--Lipid profile (mg/dl) in rabbit fed a normal (CT and R) or high cholesterol diet (CL and CR) during 60 days, with or without supplementation of resveratrol. Total Chol (mg/dL) LDL (mg/dL) Groups M SD M SD CT 47.03 (b) [+ or -] 15.84 14.88 (b) [+ or -] 20.12 R 49.51 (b) [+ or -] 17.09 12.99 (b) [+ or -] 17.38 CL 950.94 (a) [+ or -] 662.90 872.74 (a) [+ or -] 639.57 CR 1508.50 (a) [+ or -] 889.71 1407.26 (a) [+ or -] 850.93 HDL (mg/dL) Tg (mg/dL) Groups M SD M SD CT 21.12 (b) [+ or -] 5.37 55.30 (b) [+ or -] 15.55 R 26.30 (ab) [+ or -] 4.08 51.01 (b) [+ or -] 10.49 CL 39.89 (a) [+ or -] 17.43 190.57 (ab) [+ or -] 116.35 CR 40.46 (a) [+ or -] 17.00 302.43 (a) [+ or -] 175.29 Chol = cholesterol; Tg = triglycerides; M = means; SD = standard deviation. Duncan's test, P<0.05. CT = control nornolipemic; R = resveratrol normolipemic; CL = control hyperlipemic; CR = resveratrol hyperlipemic.