Elsevier

Brain Research

Volume 1107, Issue 1, 30 August 2006, Pages 192-198
Brain Research

Research Report
Quercitrin, a glycoside form of quercetin, prevents lipid peroxidation in vitro

https://doi.org/10.1016/j.brainres.2006.05.084Get rights and content

Abstract

Reactive oxygen species have been demonstrated to be associated with a variety of diseases including neurodegenerative disorders. Flavonoid compounds have been investigated for their protective action against oxidative mechanisms in different in vivo and in vitro models, which seems to be linked to their antioxidant properties. In the present study, we examine the protective mechanism of quercitrin, a glycoside form of quercetin, against the production of TBARS induced by different agents. TBARS production was stimulated by the incubation of rat brain homogenate with Fe2+, Fe2+ plus EDTA, quinolinic acid (QA), sodium nitroprusside (SNP) and potassium ferricyanide ([Fe(CN)6]3−). Quercitrin was able to prevent the formation of TBARS induced by pro-oxidant agents tested; however, it was more effective against potassium ferricyanide ([Fe(CN)6]3−, IC50 = 2.5), than quinolinic acid (QA, IC50 = 6 μg/ml) and sodium nitroprusside (SNP, IC50 = 5.88 μg/ml) than Fe2+ (Fe2+, IC50 = 14.81 μg/ml), Fe2+ plus EDTA (Fe2+ plus EDTA, IC50 = 48.15 μg/ml). The effect of quercitrin on the Fenton reaction was also investigated (deoxyribose degradation). Quercitrin caused a significant decrease in deoxyribose degradation that was not dependent on the concentration. Taken together, the data presented here indicate that quercitrin exhibits a scavenger and antioxidant role, and these effects probably are mediated via different mechanisms, which may involve the negative modulation of the Fenton reaction and NMDA receptor.

Introduction

Cell metabolism continuously produces reactive oxygen species (ROS) as by products of respiration and other metabolic activities (Azbill et al., 1997, Halliwell, 1994, Park et al., 2004). These reactive species can normally be handled by non-enzymatic and enzymatic antioxidant defenses (Rodriguez-Martinez et al., 2000, Santamaría et al., 2003). However, the imbalance between the antioxidant system and the over production of ROS has been associated with a variety of human diseases such as atherosclerosis, cancer and neurodegenerative diseases (Alexi et al., 2000, Ames et al., 1993, Johnson, 2004, Lohr, 1991, Halliwell, 1994, Witztum, 1994).

Of particular importance for the design of new therapeutic approaches, natural and synthetic antioxidant compounds can afford protection in a variety of in vitro and in vivo models of toxicity (Bastianetto and Quirion, 2002, Bellé et al., 2004, Bixby et al., 2005, Burger et al., 2004, Burger et al., 2005, Cabrera et al., 2000, Cooke et al., 2005, Dominguez et al., 2005, Ghisleine et al., 2003, Gugliucci and Stahl, 1995, Gupta et al., 2003, Moridani et al., 2003, Nogueira et al., 2004, Pérez-Severiano et al., 2004, Steffen et al., 2005, Williams et al., 2004, Youdim et al., 2004). Therefore, the consumption of foods rich in natural antioxidants is thought to be of preventive value to delay the development or to impede the manifestation of neurodegenerative diseases in humans and animal models (Aruoma et al., 2003, Naidu et al., 2003).

Flavonoids are components of the human diet and are widely found in vegetables (Hollman and Katan, 1997, Hollman and Katan, 1999) and beverages (Filip et al., 2001, Gugliucci and Stahl, 1995). Moreover, it was recently demonstrated that the plasma antioxidant status was significantly higher in animals treated with quercetin, suggesting that quercetin metabolites can retain some antioxidant activity when the o-catechol group does not undergo conjugation reactions. In the same study, the authors showed that plasma quercetin metabolites compete in vivo with other molecules for peroxynitrite (Justino et al., 2004).

Therefore, the study of the potential antioxidant activity of flavonoids has attracted the attention of researchers who intend to identify whether these compounds could be effective antioxidant agents. Although literature data indicate that different types of flavonoids are potent antioxidants, the exact mechanism(s) which underlie their protective effects is still not completely understood. There are several points of evidence in the literature indicating that polyphenols are reducing agents and free radical scavengers and they can participate in the regeneration of other antioxidants, such as vitamin E (Jovanovic et al., 1998, Rice-Evans et al., 1996).

Quercetin is the most abundant bioflavonoid found in vegetable and fruits, and this compound is mainly present in the glycoside form, for example, as quercitrin. Studies have demonstrated that the absorption of quercetin glycosides contained in onions was higher (52%) than that of quercetin aglycones (24%) (Hollman et al., 1995). In line with this, the 3-O-β-glucoside of quercetin is better absorbed than quercetin (Morand et al., 2000). Perhaps, because the glycoside form gives physical and chemical properties which are different from those of the aglycone forms. The sugar portion bound to the aglycone portion increases the solubility in polar solvents and consequently improves absorption, through the utilization of glucose transporters that are present in the intestinal mucosa (Gee et al., 1998). However, the majority of the studies have been carried out with the aglycone form and little is known about the biological properties of glycoside forms, due to the lack of commercial standards (Scalbert et al., 2002) (Fig. 1).

Quinolinic acid (QA), an endogenous tryptophan metabolite formed in the kynurenine pathway, is a potent neurotoxin and a selective NMDA subtype of the glutamate receptor agonist (Scalbert, 1993). In vitro studies have demonstrated enhanced cytosolic calcium concentrations, ATP exhaustion, GABA depletion and superoxide radical formation (Foster et al., 1983), oxidative stress and lipid peroxidation induced by QA (Bellé et al., 2004, Puntel et al., 2005, Ríos and Santamaría, 1991, Vega-Naredo et al., 2005). Behavioral alterations oxidative stress and lipid peroxidation are important features observed in vivo as a result of QA-induced neurotoxicity (Perez-de la Cruz et al., 2005, Rossato et al., 2002, Susel et al., 1989, Stýpek et al., 1997).

Sodium nitroprusside (SNP) has been suggested to cause cytotoxicity via the release of cyanide and/or nitric oxide (Bates et al., 1991, Chen et al., 1991, Dawson et al., 1991, Rauhala et al., 1998). There are several studies concerning the role of NO in the pathophysiology of strokes, traumas, seizures and Alzheimer's, and Parkinson's diseases (Bolanos and Almeida, 1999, Castill et al., 2000, Prast and Philippou, 2001, Weisinger, 2001). It is known that light exposure promotes the release of NO from SNP through a photodegradation process (Arnold et al., 1984, Singh et al., 1995), and data from the literature have demonstrated that after the release of NO, SNP or [NO–Fe–(CN)5] 2− is converted to iron containing [(CN)5–Fe]3− and [(CN)4–Fe]2− species (Loiacono and Beart, 1992). After the release of NO, the iron moiety may react with SNP, which could lead to the formation of highly reactive oxygen species, such as hydroxyl radicals via the Fenton reaction (Graf et al., 1984).

The aims of this study were to investigate the antioxidant action of quercitrin, the glycoside form of quercetin, in rat brain lipid peroxidation induced by different agents. In addition, we investigated whether the antioxidant mechanism of quercitrin involves the Fenton reaction.

Section snippets

Fe2+ and Fe2+/EDTA × quercitrin

Statistical analyses revealed that Fe2+ caused a significant stimulation of brain TBARS formation (P < 0.01), whereas quercitrin caused a reduction in the TBARS production induced by Fe2+. Fe2+ plus EDTA caused a similar increase in TBARS production to that caused by Fe2+ alone, and quercitrin was also able to reduce the stimulatory effects of Fe2+. Furthermore, quercitrin caused a significant reduction in TBARS production under basal conditions (Fig. 2).

Quinolinic acid × quercitrin

QA caused a significant increase in

Discussion

The results of the present study show that quercitrin prevented the lipid peroxidation in brain homogenates induced by different agents.

Taken together, these results can indicate that the antioxidant properties of quercitrin, a glycoside form of quercetin, depend on the agent used to induce lipid peroxidation. When SNP and QA were used to induce oxidative stress, lower quercitrin concentrations (10 μg/ml) were required to reduce TBARS to basal levels, as compared with concentrations required in

Chemicals

Tris–HCl, QA, thiobarbituric acid and malonaldehyde bis-(dimethyl acetal) (MDA) were obtained from Sigma (St. Louis, MO, USA). Sodium nitroprusside was obtained from Merck (Darmstadt, Germany). Ferrous sulphate, ethylenediamintetracetic (EDTA), hydrogen peroxide, chloridric acid and acetic acid were obtained from Merck (Rio de Janeiro, RJ, Brazil).

Quercitrin

Quercitrin (Fig. 1) was isolated from Solidago microglossa D.C. and tested at concentrations of 0–100 μg/ml. The purity of the isolated compound was

Acknowledgments

The financial support by CAPES, CNPq, FAPERGS is gratefully acknowledged.

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