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Promotion of oxidative stress by L-tryptophan

in cerebral cortex of rats
ARTICLE in NEUROCHEMISTRY INTERNATIONAL AUGUST 2006
Impact Factor: 3.09 DOI: 10.1016/j.neuint.2006.01.001 Source: PubMed

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Neurochemistry International 49 (2006) 8793

www.elsevier.com/locate/neuint

Promotion of oxidative stress by L-tryptophan in cerebral cortex of rats

Luciane Rosa Feksa a, Alexandra Latini a,b, Virgnia Cielo Rech a, Moacir Wajner a,b,
Carlos Severo Dutra-Filho a, Angela Terezinha de Souza Wyse,
Clovis Milton Duval Wannmacher a,*
a

Departamento de Bioqumica, Instituto de Ciencias Basicas da Saude, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
b
Servico de Genetica Medica, Hospital de Clnicas de Porto Alegre, Porto Alegre, RS, Brazil
Received 7 September 2005; received in revised form 5 January 2006; accepted 5 January 2006
Available online 23 February 2006

Abstract
Despite the significant brain abnormalities, the neurotoxic mechanisms of brain injury in hypertryptophanemia are virtually unknown. In this
work, it was investigated the in vitro effect of L-tryptophan on various parameters of oxidative stress, namely spontaneous chemiluminescence,
thiobarbituric acid-reactive substances (TBA-RS), total radical-trapping antioxidant potential (TRAP), total antioxidant reactivity (TAR) and
glutathione (GSH) levels in cerebral cortex from 30-day-old rats. Tryptophan significantly increased chemiluminescence and TBA-RS
measurements indicating that this amino acid induced lipid peroxidation in vitro. We also observed that tryptophan significantly decreased
the brain antioxidant defenses by reducing the values of TRAP, TAR and GSH, reflecting that the overall content of antioxidants was reduced by
tryptophan. Furthermore, the tryptophan-induced increase of TBA-RS was fully prevented by GSH and by combination of catalase plus superoxide
dismutase, but not by the inhibitor of nitric oxide synthase Nv-nitro-L-arginine methyl ester (L-NAME). In case these findings also occur in human
hypertryptophanemia or in other neurodegenerative diseases in which tryptophan accumulates, it is feasible that oxidative stress may be involved in
the mechanism leading to the brain injury observed in patients affected by these disorders.
# 2006 Elsevier Ltd. All rights reserved.
Keywords: Tryptophan; Hypertryptophanemia; Oxidative stress; Antioxidant defenses

1. Introduction
As a possible cause of the neuronal degeneration found in the
brains of patients affected by different neurological disorders,
aberrant metabolism of endogenous substances that leads to the
production of excess amounts of neurotoxic compounds has
provoked recent interest. Congenital defects of tryptophan
metabolism are comparatively rare (Snedden et al., 1983),
although at least 12 patients with apparent defects in tryptophan
(Trp) catabolism have been reported (Levy, 2001). Congenital
hypertryptophanemia is probably caused by a blockage in the
conversion of Trp to kynurenine, accumulating tryptophan and
some of its metabolites in plasma and tissues of affected patients.
Clinically, the patients present mild to moderate mental
retardation with exaggerated affective responses, periodic mood
* Corresponding author at. Departamento de Bioqumica, Instituto de Ciencias Basicas da Saude, UFRGS, Rua Ramiro Barcelos 2600, CEP 90.035-003,
Porto Alegre, RS, Brazil. Tel.: +55 51 3316 5575; fax: +55 51 3316 5535.
E-mail address: clovisdw@ufrgs.br (C.M.D. Wannmacher).
0197-0186/$ see front matter # 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.neuint.2006.01.001

swings, and apparent hypersexual behavior (Snedden et al.,

1983). Trp plasma levels, in these patients, are 10-fold increased
(up to 1 mM) and the urinary excretion of indolic acids was 100
times greater than normal. The massive excretion of the Trp
derivatives, indoleacetic, indolelactic and indolepyruvic acids,
suggested that abnormally large amounts of Trp were being
metabolized by the transamination pathway.
A number of other neurological diseases are connected with
abnormalities in the metabolism of Trp, but the relationship
between cause and effect is unclear. The kynurenine pathway,
the major metabolic pathway of the amino acid Trp has been a
focus of attention in these diseases, since this pathway produces
several compounds, namely kynurenines, serotonin, indolic
acids, nicotinic acid (niacin) compounds and quinolinic acid.
Under normal conditions, very little Trp appears to be
converted to nicotinic acid (Levy, 2001). However, patients
having a defect in Trp degradation may present clinical
characteristics of niacin deficiency such as photosensitive rash,
ataxia and mental subnormality (Wong et al., 1976; Fenton
et al., 1983; Salih et al., 1985).

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L.R. Feksa et al. / Neurochemistry International 49 (2006) 8793

Intermediate metabolites of Trp pathway have received

attention as possible responsible for neurotoxicity in diseases
affecting Trp metabolism. Quinolinic acid activates the Nmethyl-D-aspartate (NMDA) subtype of glutamate receptors
and might cause excitotoxic damage in neuronal cells (Stone
and Perkins, 1981; Schwarcz et al., 1983; Stone, 2001; Stone
and Darlington, 2002). Considering that kynurenic acid is an
antagonist of NMDA receptors (Perkins and Stone, 1982), it has
been hypothesized that these compounds, or the ratio between
them, might contribute to the neuronal dysfunction of some
common neurodegenerative diseases. Moreover, 3-hydroxykynurenine (3-HK) causes cytotoxicity and cell lysis (Eastman
and Guilarte, 1989; Eastman and Guilarte, 1990) and high
levels of this compound were found in the brain of patients with
several neurological diseases (Pearson and Reynolds, 1991;
Ogawa et al., 1992; Sardar et al., 1995).
Many neurodegenerative disorders are associated with an
excessive generation of reactive oxygen species (ROS) and
oxidative stress. The pathways to nerve cell death induced by
diverse potential neurotoxins such as peptides, excitatory
amino acids, cytokines or synthetic drugs commonly share
oxidative processes, which can cause either an acute oxidative
destruction or activate secondary events leading to apoptosis.
The oxidation of protein sulfhydryls to disulfides and their
reduction back to sulfhydryls is an early cellular response to
oxidative stress. Treatment of the oxidized protein with
glutathione may restore its enzymatic capabilities (SluisCremer et al., 1996). The pathophysiological role of ROS has
been intensively studied in vitro and in vivo models of chronic
neurodegenerative diseases such as Alzheimers disease (AD)
(Behl and Moosmann, 2002). There is growing evidence for a
degree of oxidative stress in Huntington disease (HD), which
could result from the activation of NMDA receptors or by some
other method independent of the receptors. In at least one
animal model for HD, a significant increase in lipid
peroxidation has been reported which parallels the neurological
phenotype (Perez-Severiano et al., 2000; Bogdanov et al., 2001;
Stoy et al., 2005). However, some of the oxidative Trp
metabolites of the kynurenine pathway, such as 5-hydroxytryptophan, are antioxidants (Cadenas et al., 1989).
Tryptophan loading induced oxidative stress in healthy
humans, but the oxidative stress was considered result from the
generation of quinolinic acid, 3-hydroxykynurenine, and 3hydroxyanthranilic acid, all of which are known to have the
ability to generate free radicals (Forrest et al., 2004). We have
reported that Trp, at 0.5 mM concentrations, in the range of the
levels found in the plasma of hypertryptophanemic patients,
inhibited competitively pyruvate kinase activity (Feksa et al.,
2005) and non competitively creatine kinase activity, possibly
by forming long lasting adducts with the enzyme in the brain
cortex of young rats (Cornelio et al., 2004). Therefore,
considering that oxidative stress is involved in the brain
dysfunction of various common neurodegenerative disorders
(Reznick and Packer, 1993; Halliwell and Gutteridge, 1999;
lvarez et al., 2001; Karelson et al., 2001), and Trp has
Mendez-A
not be considered an oxidant by itself, in the present study we
investigated the in vitro effect of Trp on various parameters of

oxidative stress in cerebral cortex from young rats, in the hope

to contribute to the understanding of the mechanisms of brain
damage in diseases affecting the metabolism of this amino acid.
2. Material and methods
2.1. Animals and reagents
Male Wistar rats of 30 days of life obtained from the Central
Animal House of the Department of Biochemistry, ICBS,
Universidade Federal do Rio Grande do Sul, Porto Alegre, RS,
Brazil, were used. The animals were maintained on a 12:12 h
light/dark cycle (lights on 07.0019.00 h) in conditioned
constant temperature (22 8C
1 8C) colony room, with free
access to water and a commercial chow (Germani, Porto
Alegre, RS, Brazil) containing 20.5% protein (predominantly
soybean supplemented with methionine), 54% carbohydrate,
4.5% fiber, 4% lipids, 7% ash and 10% moisture. The
Principles of laboratory animal care (NIH publication no 8023, revised 1996) were followed in all experiments and our
research protocol was approved by the Ethical Committee for
animal experimentation of the Universidade Federal do Rio
Grande do Sul. All efforts were made to minimize animal
suffering and to use only the number of animals necessary to
produce reliable scientific data. All chemicals were purchased
from Sigma (St. Louis, MO, USA), except thiobarbituric acid,
which was purchased from Merck (Darmstadt, Germany) and
superoxide dismutase, which was obtained from RANDOX
(Antrim, United Kingdom). Trp was prepared on the day of the
experiments in 20 mM sodium phosphate buffer, pH 7.4
containing 140 mM KCl. The final concentrations of Trp in the
medium were 1, 2.5 and/or 5 mM. Chemiluminescence was
assayed using a Tri-Carb 2100TR beta liquid scintillation
spectrometer, and TRAP and TAR using a Wallac 1409
scintillation counter. TBA-RS and GSH levels were measured
with a spectrophotometer with temperature control (Hitachi U2001) and a fluorescence spectrophotometer (Hitachi F 2000),
respectively.
2.2. Tissue preparation and incubation
On the day of the experiments the rats were sacrificed by
decapitation without anesthesia, and the brain was rapidly
excised on a Petri dish placed on ice. The olfactory bulbs, pons,
medulla and cerebellum were discarded and the cerebral cortex
was dissected, weighed and kept chilled until homogenization.
Brain tissue was homogenized in 10 volumes (1:10, w/v) of
20 mM sodium phosphate buffer, pH 7.4 containing 140 mM
KCl. Homogenates were centrifuged at 750  g for 10 min at
4 8C to discard nuclei and cell debris (Llesuy et al., 1985;
Gonzalez-Flecha et al., 1991). The pellet was discarded and the
supernatant, a suspension of mixed and preserved organelles,
including mitochondria, was separated and immediately used
for the various analyses.
Cerebral cortex supernatants were incubated for 090 min at
37 8C in the presence of Trp at concentrations ranging from 1,
2.5 to 5 mM. Controls did not contain Trp in the incubation

L.R. Feksa et al. / Neurochemistry International 49 (2006) 8793

89

medium. Immediately after incubation, aliquots were taken to

measure chemiluminescence, TBA-RS, TRAP, TAR and GSH.
In some experiments antioxidants were also added to the
incubation medium at the following concentrations: 200 mM
GSH, 500 mM Nv-nitro-L-arginine methyl ester (L-NAME) and
50 mU/mL catalase and 50 mU/mL superoxide dismutase.

called induction time (IT). IT is directly proportional to the

antioxidant capacity of the tissue and the IT of each sample was
compared with the IT of standard trolox. TRAP values were
calculated as nmol trolox/mg protein.

2.3. Thiobarbituric acid-reactive substances (TBA-RS)

TAR, which represents the quality of the tissue antioxidants,

was determined by measuring the luminol chemiluminescence
intensity induced by AAPH according to the method of Lissi
et al. (1995). The background chemiluminescence was
measured by adding 4 mL 2 mM ABAP (in 0.1 M glycine
buffer, pH 8.6) into a glass scintillation vial. Fifteen microliter
of luminol (4 mM) were added to each vial and the
chemiluminescence was measured. This was considered to
be the basal value. Ten microliter of 10 mM trolox or tissue
supernatant was then added and the chemiluminescence was
measured over 60 s. The trolox or supernatant addition reduces
the chemiluminescence. The rapid reduction in luminol
intensity is considered as a measure of its TAR capacity.
TAR measurement was calculated as nmol trolox/mg protein.

TBA-RS was determined according to the method described

by Esterbauer and Cheeseman (1990). Briefly, 300 mL of cold
10% trichloroacetic acid were added to 150 mL of pre-treated
cortical supernatants and centrifuged at 300  g for 10 min.
Three hundred microliter of the supernatant were then
transferred to a pyrex tube and incubated with 300 mL of
0.67% thiobarbituric acid in 7.1% sodium sulphate in boiling
water bath for 25 min. The mixture was allowed to cool on
water for 5 min. The resulting pink stained TBA-RS was
determined in a spectrophotometer at 535 nm. Trp did not
produce colour when tested without the addition of the
supernatant, demonstrating the absence of a direct reaction to
thiobarbituric acid. Calibration curve was performed using
1,1,3,3-tetramethoxypropane and each curve point was subjected to the same treatment as that of the supernatants. TBARS was calculated as nmol TBA-RS/mg protein.
2.4. Chemiluminescence
Chemiluminescence, the spontaneous light emission mainly
from peroxidizing lipids, was assayed in a dark room by the
method of Gonzalez-Flecha et al. (1991). Incubation flasks
containing 3.5 mL of 20 mM sodium phosphate buffer, pH 7.4
containing 140 mM KCl were counted for background
chemiluminescence during 5 min. An aliquot of 0.5 mL of
pre-treated cortical supernatants was then added and the
chemiluminescence was measured for further 30 min at room
temperature. The background chemiluminescence was subtracted from the total value. Chemiluminescence was calculated
as cpm/mg protein and represented as percentage of controls.
2.5. Total radical-trapping antioxidant potential (TRAP)
TRAP, representing the total non-enzymatic antioxidant
capacity of the tissue, was determined by measuring the luminol
chemiluminescence intensity induced by 2,20 -azo-bis-(2-amidinopropane) (AAPH) according to the method of Lissi et al.
(1992). The background chemiluminescence was measured by
adding 4 mL of 10 mM ABAP dissolved in 0.1 M glycine buffer,
pH 8.6, into a glass scintillation vial. Ten microliter of luminol
(4 mM) were added to each vial and the chemiluminescence was
measured. This was considered to be the initial value. Ten
microliter of 300 mM trolox (water-soluble a-tocopherol) or
tissue supernatant was added and the chemiluminescence was
measured until it reached the 30% of initial chemiluminescence
levels. The trolox or supernatant addition to the incubation
medium reduces the chemiluminescence. The time necessary for
the chemiluminescence intensity returns to the initial value is

2.6. Total antioxidant reactivity (TAR)

2.7. Glutathione levels (GSH)

GSH levels were measured according to Browne and
Armonstrong (1998). Pre-treated cerebral cortex supernatants
were diluted in 20 volumes (1:20, v/v) of 100 mM sodium
phosphate buffer pH 8.0, containing 5 mM EDTA. One hundred
microliter of this preparation were incubated with an equal
volume of O-phthaldialdehyde (1 mg/mL methanol) at room
temperature during 15 min. Fluorescence was measured using
excitation and emission wavelengths of 350 and 420 nm,
respectively. Calibration curve was performed with standard
GSH (0.011 mM) and the concentration was calculated as
pmol/mg protein.
2.8. Protein determination
Protein concentrations were determined in cerebral cortex
supernatants by the method of Lowry et al. (1951), using bovine
serum albumin as standard.
2.9. Statistical analysis
Data were analyzed by analysis of variance (ANOVA)
followed by the Tukey test when the F value was significant.
Linear regression analysis was also used to test concentrationor time-dependent effects. All analyses were performed using
the Statistical Package for the Social Sciences (SPSS) software
in a PC-compatible computer. A value of P < 0.05 was
considered to be significant.
3. Results
Initially, we investigated the in vitro effect of Trp on the lipid
peroxidation parameter TBA-RS measurement at different
incubation times in rat cerebral cortex supernatants. Fig. 1

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L.R. Feksa et al. / Neurochemistry International 49 (2006) 8793

Fig. 1. Effect of tryptophan (Trp) on thiobarbituric-acid reactive substances

(TBA-RS) in rat cerebral cortex at different pre-incubation times. Bars are
mean
S.D. for six independent experiments (animals) performed in triplicate.
Different from control, * p < 0.05; **p < 0.01 (Tukey test).

shows that Trp significantly increased TBA-RS values after 30

(42%), 60 (38%) and 90 (36%) min of pre-incubation [30 min:
F (3,20) = 8.52; P < 0.001; 60 min: F (3,20) = 5.37; P < 0.01;
90 min: F (3,20) = 5.62; P < 0.01] and that the effect was dosedependent [30 min: b = 0.71; P < 0.001; 60 min: b = 0.63;
P < 0.001; 90 min: b = 0.67; P < 0.001]. In addition, it can
also be observed in the figure that TBA-RS was also increased
by 2.5 mM Trp (30%) at longer incubation times. As similar
Trp effects were found at 60 and 90 min of pre-incubation, we
set up 1 h as standard incubation time for assessing spontaneous
chemiluminescence. Fig. 2 shows that chemiluminescence was
significantly increased (up to 45%) by Trp [F (3,16) = 4.65;
P < 0.05] in a concentration dependent manner [b = 0.81;
P < 0.001]. These results strongly indicate that lipid peroxidation is elicited by Trp in brain tissue.
Next, we evaluated the role of antioxidants on the effect
produced by Trp on TBA-RS measurement. Rat cortical

Fig. 2. Effect of tryptophan on chemiluminescence in rat cerebral cortex. Bars

are mean
S.D. for five independent experiments (animals) performed in
duplicate and are represented as percentage of controls (1635
350 cpm/mg
protein). Different from control, *p < 0.05; **p < 0.01 (Tukey test).

Fig. 3. Effect of antioxidants on tryptophan (Trp)-induced increase of thiobarbituric-acid reactive species (TBA-RS) in rat cerebral cortex. A: glutathione
(GSH; 200 mM) and Nv-nitro-L-arginine methyl ester (L-NAME; 500 mM); B:
catalase (CAT; 50 mU/mL; superoxide dismutase (SOD; 50 mU/mL). Bars are
mean
S.D. for seven independent experiments (animals) performed in triplicate
and are expressed as nmol/mg protein. Different from control, *p < 0.05; ***p <
0.001. Different from Trp 2.5 mM, # p < 0.05; ## p < 0.01; ###p < 0.001(Tukey
test).

supernatants were incubated with 2.5 mM Trp alone or in the

presence of the free radical scavengers GSH, L-NAME, CAT,
SOD or combination of SOD plus CAT. Fig. 3A shows that the
Trp-induced in vitro lipid peroxidation was significantly
attenuated (10%) by 200 mM GSH, in contrast to 500 mM LNAME which did not prevent the increased TBA-RS values
[F (5,36) = 88.43; P < 0.001]. Similar results were achieved with
the combination of 50 mU/mL SOD plus 50 mU/mL catalase
(15%) (Fig. 3B) but not when the enzymes were added
separately to the supernatants [F (7,40) = 3.08; P < 0.05]. The
data suggest the probable participation of superoxide and
hydroxyl radicals in Trp-induced lipid peroxidation.
Then, the effects of Trp on the brain non-enzymatic
antioxidants defenses were investigated. Fig. 4A shows that
TRAP measurement, which is indicative of the overall content
of brain non-enzymatic antioxidants, was significantly reduced
(up to 45%) by Trp [F (3,16) = 7.31; P < 0.05] in a dose
dependent manner [b = 0.75; P < 0.001]. Moreover, TAR
measurement, which is a measure of the tissue capacity to react
with free radicals, was also markedly reduced (up to 62%) by
Trp even at the lowest dose tested [F (3,16) = 35.34; P < 0.001]

L.R. Feksa et al. / Neurochemistry International 49 (2006) 8793

91

These findings suggest that the overall content of the nonenzymatic antioxidants in cerebral cortex as well as the brain
capacity to modulate the damage associated with enhanced free
radical production were significantly reduced by Trp treatment.
4. Discussion

Fig. 4. Effect of tryptophan on total radical-trapping antioxidant potential

(TRAP) (A), total antioxidant reactivity (TAR) (B), and glutathione levels
(GSH) and (C) in rat cerebral cortex. Bars are mean
S.D. for five to six
independent experiments (animals) performed in duplicate. Different from
control, *p < 0.05; **p < 0.01; ***p < 0.001 (Tukey test).

and in a concentration-dependent manner [b = 0.92;

P < 0.001] (Fig. 4B). Finally, we assessed the Trp in vitro
effect on the tissue levels of GSH, the major contributor to the
TRAP value in the brain. It is shown in Fig. 4C that GSH levels
were significantly reduced (up to 80%) [F (3,16) = 37.95;
P < 0.001] in a concentration-dependent manner [b = 0.81;
P < 0.0001] when cortical supernatants were exposed to Trp.

Tryptophan was considered an antioxidant agent (Watanabe

et al., 2002), but urate, a known antioxidant agent, may be
oxidized by tryptophan neutral radical (Santus et al., 2001).
However, when oxidized, the indoxyl radical formed from
tryptophan subsequently oxidizes a tyrosine side chain of
lysozymes to the phenoxyl radical (Stuart-Audette et al., 2003).
Considering that Trp inhibits the in vivo activity of creatine
kinase from rat brain (Cornelio et al., 2004), and Trp
administration induces oxidative stress (Forrest et al., 2004),
in the present study we investigated the in vitro effect of Trp on
the lipid peroxidation parameters spontaneous chemiluminescence and thiobarbituric acid-reactive substances (TBA-RS)
and on the non-enzymatic antioxidant defenses, the total
radical-trapping antioxidant potential (TRAP), the total
antioxidant reactivity (TAR) and glutathione (GSH) levels in
cerebral cortex from young rats.
We observed that Trp significantly increased TBA-RS values
in a dose-dependent manner and also in a time-dependent
manner showing the highest in vitro effect after 1 h preincubation of brain supernatants with the amino acid. Therefore, this exposure time was set up as the standard Trpexposition time to assess the other parameters of oxidative
stress. Thus, we observed that Trp also increased chemiluminescence in brain supernatants. As light emitted in the
chemiluminescence assay usually arises from peroxidizing
lipids due to an increase in reactive oxygen or nitrogen species
production and that TBA-RS reflects the amount of malondialdehyde formation, an end product of membrane fatty acid
peroxidation (Halliwell and Gutteridge, 1999), our data
indicate that Trp-induced lipid peroxidation in rat cerebral
cortex probably occurs due to free radical generation.
We also observed a significant attenuation of the Trp-elicited
increase of TBA-RS measurement when cerebral cortex
supernatants were exposed to the free radical scavenger
GSH or the combination of CAT plus SOD, indicating that
superoxide and hydroxyl radicals might be involved in Trp
effect. In contrast, when SOD or CAT were supplemented
separately to the supernatants, they were not able to reduce the
increase of TBA-RS provoked by Trp, suggesting that hydrogen
peroxide did not participate in the events induced by the acid.
Similarly, the nitric oxide synthase inhibitor L-NAME did not
prevent the increased TBA-RS levels; in contrast it magnified
these values. In this particular, these results are in agreement
with previous studies stating the concept that nitric oxide is a
potent chain breaking antioxidant toward peroxidizing lipids,
due to facile radical-radical termination reactions with lipid
radical species (lipid peroxides), thus preventing a-tocopherol
oxidation (Rubbo et al., 2000; Zhu and Fung, 2000). Therefore,
the inhibition of lipid peroxidation propagation reactions by
using nitric oxide synthesis inhibitor would lead to increased

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L.R. Feksa et al. / Neurochemistry International 49 (2006) 8793

TBA-RS levels as observed in Fig. 3A. On the other hand, since

the prevention of Trp-induced lipid peroxidation was only
partial, it is feasible that other reactive species not scavenged by
the antioxidants used in our assays were involved in this effect,
or long lasting adducts occurred.
Next, the analysis of the non-enzymatic antioxidant defenses
revealed that Trp markedly reduced the capacity to handle an
increased free radicals (TAR measurement) and severely
diminished the overall content of non-enzymatic antioxidants
(TRAP values) and GSH, the main natural-occurring antioxidant. It can be concluded that the rat cortical antioxidant
defenses are severely compromised by Trp since these three
parameters can be used as an index of the capacity of a tissue to
prevent the damage associated to free radical processes (Lissi
et al., 1995; Halliwell and Gutteridge, 1999; Evelson et al.,
2001). It might be also presumed that the decreased of the brain
antioxidant defenses was secondary to an increase of reactive
species particularly superoxide and hydroxyl radicals, since the
antioxidants used to prevent the Trp-induced TBA-RS levels
were able to scavenge these free radicals.
Taken together, it seems reasonable to propose that Trp
provokes a significant in vitro stimulation of oxidative stress in
rat cerebral cortex, a deleterious cell condition, which induces
oxidation of proteins, lipids, sugars and DNA (Halliwell and
Gutteridge, 1999). However, we cannot exclude the possibility
that metabolites from Trp could account, at least in part, for the
effects observed in this study.
At this point, it should be emphasized that the brain has low
cerebral antioxidant defenses compared with other tissues
(Halliwell and Gutteridge, 1996), a fact that makes this tissue
more vulnerable to increased reactive species. It is also feasible
that oxidative stress may act synergistically with other
pathophysiologic mechanisms, such as excitotoxicity as well
as energy metabolism impairment, to accelerate brain damage
in these neurodegenerative disorders. Finally, it is proposed that
the administration of antioxidants as well as chelators of
transition metal ions should be considered as an adjuvant
therapy to specific diets or to other pharmacological agents for
these patients
Acknowledgements
This work was supported in part by grants from Fundacao de
Amparo a Pesquisa do Rio Grande do Sul (FAPERGS),
Conselho Nacional de Desenvolvimento Cientfico e tecnologico (CNPq), Programa de Nucleos de Excelencia Financiadora
de Estudos e projetos (PRONEX).
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