Original Article
SELECTIVE NF-κB INHIBITORS PREVENT NEPHROTOXICITY OF GENTAMICIN
TUGCU
et al.
Selective nuclear factor k-B inhibitors, pyrolidium
dithiocarbamate and sulfasalazine, prevent the
nephrotoxicity induced by gentamicin
VOLKAN TUGCU, EMIN OZBEK*, ALI I. TASCI, ERAY KEMAHLI, ADNAN SOMAY†, MUZAFFER BAS, CETIN KARACA‡,
TUNCAY ALTUG‡, MUSTAFA B. ÇEKMEN¶ and HACI K. ÖZDOGAN¶
Department of Urology, Bakyrköy Dr. Sadi Konuk Research and Training Hospital, Departments of *Urology and †Pathology, Vakyf Gureba Research
and Training Hospital, ‡ Animal Research Laboratory, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, and ¶Biochemistry, Medical Faculty,
Kocaeli University, Kocaeli, Turkey
Accepted for publication 28 March 2006
OBJECTIVE
To investigate the effect of selective nuclear
factor κ-B (NFκ-B) inhibitors, pyrolidium
dithiocarbamate (PD) and sulfasalazine (SZ)
on renal tubular necrosis and inducible nitric
oxide synthase (iNOS) and NFκ-B expression
induced by gentamicin in rats.
MATERIALS AND METHODS
In all, 48 adult male Sprague-Dawley rats
were divided into six equal groups; group 1,
control; group 2, injected with gentamicin for
10 days (100 mg/kg/day, intraperitoneal, i.p.);
group 3, injected with gentamicin plus PD
(100 mg/kg/day, i.p.); group 4, injected with
gentamicin plus SZ (75 mg/kg/day, i.p.); group
5, injected with gentamicin plus distilled
water (vehicle for PD); and group 6, injected
with gentamicin plus ammonium hydroxide
(75 mg/day, 1 M, vehicle for SZ) for 10 days. At
24 h after the last injection, rats were killed
and the renal cortex separated from the
INTRODUCTION
Aminoglycoside antibiotics are often essential
for treating severe infections by Gram-
negative bacteria, but they cause nephrotoxic
reactions [1]. There is increasing evidence
suggesting that gentamicin-induced
glomerular dysfunction in vivo is mediated
by reactive oxygen species (ROS), as giving
antioxidants attenuates the decrease in GFR
[2,3]. In vitro experiments showed that
gentamicin enhances ROS production and
that renal cortical mitochondria are the
source of ROS [4]. The same authors showed
that ROS could be responsible for proximal
tubular necrosis and acute renal failure
caused by gentamicin in vivo [5].
680
medulla. A small sample was fixed in
formaldehyde solution for histological and
immunohistochemical examination. Blood
samples were also taken to assess the
serum levels of urea, creatinine, Na+, K+ and
γ-glutamyl transpeptidase (GT). Crude
extracts of the cortex were used to
determine reduced glutathione (GSH-Px),
NO and malondialdehyde (MDA).
Immunohistochemically, iNOS and the active
subunit of NFκB, P65, were evaluated using
mouse monoclonal antibodies.
RESULTS
On haematoxylin and eosin staining,
compared with the controls rats, gentamicin
caused widespread tubular necrosis (grade 3
and 4) but in group 3 and 4 there was a
marked reduction in the extent of tubular
damage. Immunohistochemically there was
more marked staining for iNOS and P65
expression in rats given gentamicin than in
the control and group 3 and 4 (P < 0.001). In
Gentamicin causes morphological and
functional changes in renal cortical
mitochondria that occur before renal
tubular cell injury or necrosis is induced by
gentamicin [6]. The specificity of gentamicin
for renal toxicity is apparently related to
its preferential accumulation in the renal
proximal convoluted tubules (50–100 times
more than in serum) [7]. Iron chelators and
free radical scavengers are reportedly
protective against gentamicin nephrotoxicity
[8–10].
Nuclear factor κ-B (NF-κB) regulates host
inflammatory and immune responses [11,12]
and cellular growth properties [13] by
increasing the expression of specific cellular
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A1
groups 3 and 4 iNOS and P65 expression were
significantly less than in rats given only
gentamicin. There was no significant
difference in serum levels of Na+, K+, blood
urea nitrogen and creatinine. Compared
with control rats, gentamicin caused
hyperproteinuria, a marked increase in levels
of serum γ-GT, MDA and NO, and a decrease in
GSH-Px (P < 0.001).
CONCLUSION
These results indicate that gentamicin induces
iNOS expression through activation of NFκ-B
(P65). It is possible to prevent gentamicin-
induced nephrotoxicity using selective NFκ-B
inhibitors.
KEYWORDS
selective nuclear factor κB, pyrolidium
dithiocarbamate, sulfasalazine, gentamicin,
nephrotoxicity, iNOS
genes. These include genes encoding at least
27 different cytokines and chemokines,
receptors involved in antigen presentation,
and receptors required for neutrophil
adhesion and migration [14]. Cytokines that
are stimulated by NF-κB, e.g. interleukin-1β
and TNF-α, can also directly activate the
NF-κB pathway, thus establishing a positive
autoregulatory loop that can amplify the
inflammatory response and increase the
duration of chronic inflammation.
It was suggested that ROS such as hydroxyl
radicals and superoxide anions act as
mediators of NF-κB activation by F-κB
degradation [15–17]. Excessive nitric oxide
(NO) production due to elevated expression of
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 SELECTIVE NF-κB INHIBITORS PREVENT NEPHROTOXICITY OF GENTAMICIN
inducible NO synthase (iNOS) might impose
cytotoxic effects on various organs, including
the kidney [18,19]. NO at high levels can
rapidly react with superoxide anion to yield a
potent antioxidant, peroxynitrite (OONO–),
which in turn causes extensive protein
tyrosine nitration [20]. The expression of
iNOS is mainly controlled by the activation
of its transcriptional factors, including
NF-κB [20].
Pyrrolidine dithiocarbamate (PD) is a metal
chelator and antioxidant, with potent anti-
inflammatory properties, in part due to its
ability to suppress NF-κB [21]. Sulfasalazine
(SZ) is an anti-inflammatory agent that is
used in the management of inflammatory
bowel disease and rheumatoid arthritis.
Treatment of colonic epithelial cells with
SZ, but not 5-aminosalicylic acid or
sulfapyride, inhibits NF-κB activation
induced by treatment with either TNF-α,
lipopolysaccharide or phorbol ester [22]. The
inhibition of the NF-κB pathway by SZ is
associated with suppression of I(κ)β(α)
phosphorylation and its subsequent
degradation [23].
Glutathione peroxidase (GSH-Px) catalyses
the reduction of hydrogen peroxide and a
variety of organic hydroperoxides by
glutathione, and is thought to participate in
the protection of cell membranes against lipid
peroxidation [24]. The kidney is important in
selenium homeostasis and in the synthesis of
plasma GSH-Px [25]. The activity of GSH-Px in
blood and tissues is related to the selenium
concentration that catalyses the reduction of
hydroxyperoxides. Its main function is to
protect against the damaging effect of
endogenously formed hydroxyperoxides
Malondialdehyde (MDA) is a physiological
compound produced by peroxidative
decomposition of unsaturated lipids as a by-
product of arachidonic acid metabolism.
Under normal conditions, MDA is quickly
oxidized to acetate or malonate and then to
CO2 via the tricarboxylic acid cycle. MDA is
one of the important markers of lipid
peroxidation [26]. Excessive production of
MDA, as a result of tissue injury and DNA
damage, could combine with free amino
groups of proteins, resulting in MDA-modified
protein adducts.
Thus the purpose of the present study was to
examine antioxidant enzyme activities and
tubular necrosis in rat kidneys treated with
© 2006 THE AUTHORS
JOURNAL COMPILATION © 2006 BJU INTERNATIONAL
gentamicin, and compare the results with
those from rats treated with PD and SZ.
MATERIALS AND METHODS
Adult male Sprague-Dawley rats (230–250 g)
were acquired from the Experimental Animal
Laboratory of Istanbul University, Cerrahpasa
Medical Faculty vivarium sources, and
maintained in a 14-h light/10-h dark cycle
with free access to food and water. In all, 48
rats were divided into six equal groups: group
1, control; group 2, injected with gentamicin
(Deva Corp., Ankara, Turkey) for 10 days
(100 mg/kg/day, i.p.); group 3, injected with
gentamicin plus PD (Sigma-Aldrich Chemical
Corp, MO, USA; 100 mg/kg/day, i.p); group 4,
injected with gentamicin plus SZ (Sigma-
Aldrich; 75 mg/kg/day, i.p.); group 5, injected
with gentamicin plus distilled water (vehicle
of PD); and group 6, injected with gentamicin
plus ammonium hydroxide (75 mg/kg/day,
1 M NH3, vehicle of SZ) for 10 days. At 24 h
after the last injection, rats were killed
under anaesthesia (i.p. sodium pentobarbital,
50 mg/kg body weight). The kidneys were
quickly removed and decapsulated, and
the renal cortex carefully separated from
the medulla, then homogenized as
described previously [2]. Small samples
were fixed in formaldehyde solution for
histopathological and immunohistochemical
examination.
For the histopathological examination the
tissues were prepared for routine examination
by light microscopy, after staining with
haematoxylin and eosin (H&E). The kidney
sections were analysed semi-quantitatively
using the technique of Houghton et al. [27].
The changes were limited to the tubulo-
interstitial areas and graded as: 0, normal; 1,
areas of focal granulo-vacuolar epithelial cell
degeneration and granular debris in the
tubular lumen, with or with no evidence of
tubular epithelial cell desquamation in small
foci (<1% of the tubule population involved
by desquamation); 2, tubular epithelial
necrosis and desquamation easily visible, but
involving less than half of the cortical tubules;
3, more than half of the proximal tubules
showing desquamation and necrosis, but
involved tubules easily visible; 4, complete or
almost complete proximal tubular necrosis.
For the immunohistochemical evaluation,
specimens were processed for light
microscopy and sections incubated at 60 °C
overnight and then de-waxed in xylene for
30 min. After rehydrating in a decreasing
series of ethanol, sections were washed with
distilled water and PBS for 10 min. Sections
were then treated with 2% trypsin in 50 mM
Tris buffer (pH 7.5) at 37 °C for 15 min, and
washed with PBS. Sections were delineated
with a Dako pen (Dako, Glostrup, Denmark)
and incubated in a solution of 3% H2O2 for
15 min to inhibit endogenous peroxidase
activity. Then, sections were incubated with
NF-κB/P65 (Rel A) Ab-1 (R-B-1638-R7,
Neomarkers, Labvision, Fremont, CA, USA) and
iNOS Ab-1 (R-B-1605-R7, Neomarkers)
antibodies. The Ultra-vision (Labvision)
horseradish peroxidase/3-amino-9-
ethylcarbazole staining protocol was used at
this stage. Sections prepared for each case
were examined by light microscopy. Sections
of rat lung were used as the control for
immunohistochemical staining specificity,
according to data provided by the antibody
manufacturer.
The sections were evaluated for diffuseness
and staining. Renal parenchymal changes,
which were evaluated according to the
diffuseness and intensity of staining in
tubulo-interstitial regions. For staining
diffuseness, sections were graded as: 0, no
staining; 1, staining <25%; 2, staining 25–
50%; 3, staining 50–75%; 4, staining >75%.
For staining intensity, sections were graded
as: 0, no staining; 1, weak but detectable
above control; 2, distinct; 3, intense.
Immunohistochemical values were obtained
by adding the diffuseness and intensity
scores, and the results compared using the
chi-square test.
For biochemical determinations, all tissues
were washed twice with cold saline solution
and immediately stored at −80 °C for
measuring MDA, GSH-Px and NO levels.
Tissues were homogenized in a four volumes
of ice-cold buffer containing 20 mM Tris,
10 mM EDTA (pH 7.4). Total nitrite (NOx)
was quantified by the Griess reaction [28,29]
after incubating the supernatant with
Escherichia coli nitrate reductase to
convert NO3 to NO2 [28,29]. Griess reagent
(1 mL 1% sulfanilamide, 0.1% naphthyl-
ethylenediamine hydrochloride, and 2.5%
phosphoric acid; Sigma Chemical Co., St.
Louis, MO, USA) was then added to 1 mL of
supernatant. The absorbance was read at
545 nm after a 30-min incubation. The
absorbance was compared with the standard
graph of NaNO2, obtained from the reduction
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T U G C U ET AL.
of NaNO3 (1–100 µmol/L). The accuracy of the
assay was checked in two ways; the inter- and
intra-assay coefficients of variation were
7.52% and 4.61%, respectively. To check
conversion of nitrate to nitrite (recovery rate),
known amounts of nitrate were added to
control plasma samples; these samples were
deproteinised and reduced as above. The
recovery rate of nitrate was 98.3 ± 7.09%.
Thiobarbituric acid (TBA) reacts with
lipoperoxidation aldehydes, e.g. MDA, as
the most common method to assess lipid
peroxidation in biological samples. The
procedure was modified from Buege and Aust
[30]. Briefly, 0.5 mL tissue homogenate was
added to a reaction mixture (1.0 mL) formed
by equal parts of 15% trichloroacetic acid,
0.25 M HCl, and 0.375% TBA, plus 2.5 mM
butylated hydroxytoluene (BHT) and 0.1 mL
of 8.1% SDS, followed by 30 min heating
at 95 °C; the pH of the analytical reaction
mixture was ≈0.9. BHT was used to prevent
lipid peroxidation during heating. After
cooling the chromogen was extracted with
n-butanol and read spectrophotometrically at
532 nm against a reaction mixture ‘blank’
lacking plasma but subjected to the entire
procedure and extracted with n-butanol.
To correct for background absorption,
absorbance values at 572 nm were subtracted
from those at 532 nm, the latter representing
the absorption maximum of the 2 : 1
TBA : MDA adduct [31]. A molar extinction
coefficient of 154 000 was used.
GSH-Px levels were determined by the
spectrophotometric method of Ellman [32]
based on the development of a yellow
solution when 5,5′ dithiobis-2-nitrobenzoic
acid is added to compounds containing
sulphydryl groups; the result are expressed as
nmol/g wet tissue.
Blood samples were also taken to assess
the serum levels of urea, creatinine, Na+,
K+ and γ-glutamyl transpeptidase (GT).
Before death a urine sample was assayed
for protein. All biochemical variables
were determined using an Olympus
Autoanalyser (Olympus Instruments,
Tokyo, Japan).
Statistical analyses of the histopathology
and immunohistochemistry results among
the groups was by the chi-square test,
with biochemical values compared using
the Mann–Whitney U-test, with P < 0.05
considered to indicate statistical significance.
682
RESULTS
There were no significant differences in blood
urea nitrogen, serum creatinine, K+ and Na+
levels among the six groups. Serum γ-GT and
urinary protein levels were higher in groups 2,
5 and 6 than in groups 1, 3 and 4 (P < 0.001;
Table 1). Groups given NFκ-B inhibitors (PD
and SZ) and gentamicin (groups 3 and 4) had
significantly lower MDA and NO levels and
higher GSH-Px levels in kidney cortex tissue
than those given gentamicin alone (group 2;
Table 1; P < 0.001).
Figure 1A shows the normal appearance of
the kidney on light microscopy (group 1,
control). In groups 2, 5 and 6 the tubular
necrosis was grade 3 or 4 (Fig. 1B,C) but
in groups 3 and 4 there was a marked
reduction in the extent of tubular damage
(Fig. 2). The semiquantitive analysis of
renal histology is also shown in Table 1. On
immunohistochemical evaluation there was
more intense expression of iNOS and p65 in
renal cortex within proximal tubules in groups
2, 5 and 6 than in groups 1, 3 and 4 (Table 1,
Fig. 2; P < 0.001).
DISCUSSION
Several hypotheses were suggested to
elucidate the possible mechanisms involved
in the tubular and glomerular effects of
gentamicin, one proposed mechanism
including the production of ROS [2,33,34].
Rivas-Canebero et al. [35] showed greater NO
production in mesangial cells incubated with
gentamicin. RO intermediates are also
involved in NF-κB activation in many studies
[15,16,36]. Animal studies showed that
endotoxaemia [37], blood loss [38–40] and
hyperoxia [41–43] all result in increased NF-
κB activation. In the present study we sought
to determine if iNOS is activated by NF-κB in
gentamicin nephrotoxicity.
NO is important in a variety of physiological
and pathophysiological processes in the
kidney [43,44]. NO can interact with ROS to
form peroxynitrite, which induces protein
damage by forming nitrotyrosine. It is
generally thought that the endothelial NOS-
derived NO, at low levels, regulates the
physiological vasodilatation, while excess NO
production due to elevated expression of
iNOS can cause cytotoxic effects in
surrounding cells. Renal function can be
perturbed by changes in NO production in the
JOURNAL COMPILATION
kidney. The contribution of NO to tissue injury
can be a direct effect mediated by NO itself
[45] and an indirect effect mediated by
reactive nitrogen species produced by the
interaction of NO with superoxide anions or
oxygen [45,46]. For example, iNOS-mediated
NO production was significantly elevated
during renal ischaemia/reperfusion injury
[18,43,44,47,48].
NF-κB was suggested to mediate
lipopolysaccharide and γ-interferon induction
of NOS in rat alveolar macrophages [49] and
murine bone marrow-derived macrophages
[50]. Furthermore, Xie et al. [17] reported the
presence of potential NF-κB binding sites in
the 5′-flanking region of the iNOS gene.
The expression of iNOS is mainly controlled by
activation of its transcriptional factors,
including NF-κB. Zhang et al. [51] reported
that homocysteine, at pathophysiological
concentrations, was able to activate NF-κB,
causing enhanced iNOS expression in
macrophages. Leung et al. [52] showed that
urinary NO was dramatically increased in rats
given gentamicin in the short term.
We showed immunohistochemically that
iNOS expression was increased in rats given
gentamicin only, but iNOS expression was
decreased in rats given gentamicin and NF-κB
inhibitors. The present study suggests that
iNOS expression can be inhibited by NF-κB
antagonism.
The NF-κB transcription factors are critical for
activating genes involved in immunological,
inflammatory and growth-related processes
[53,54]. The present results provide evidence
that activating NF-κB and iNOS could be
important in the progression of cortical
tubular injury.
NF-κB proteins are composed of at least
seven members: NF-κB1 (p50/105), NF-κB2
(p52/100), c-rel, rel-A (p65), relB, v-Rel and
the Drosophila Rel proteins [53,54]. We
focused on p65 because of its known
importance for positive transcriptional
regulators and their role in the production of
pro-inflammatory mediators [53,55–57].
Saura et al. [57] reported that p65 and p50
were predominant in the production of nitrite
in rat mesangial cells. Nakashima et al. [58]
reported that inhibition of active NF-κB by
dexamethasone, acetylsalicylic acid or PD was
caused by suppression of only the p65 subunit
of NF-κB. The p50 subunit was constitutively
© 2006 THE AUTHORS
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 SELECTIVE NF-κB INHIBITORS PREVENT NEPHROTOXICITY OF GENTAMICIN

TABLE 1 The biochemical effects, histological scores and histopathological effects of gentamicin, SZ and PD in rats

Group
Variable 1 2 3 4 5 6
Urea, mg/L 140 160 151 154 158 160
Creatinine, mg/L 8 9 8.6 8.8 9.2 9.1
SU+, mmol/L 138 143 141 140 143 143
K+, mmol/L 3.8 4.3 4.2 4.1 4.4 4.3
γ-GT, IU/L 542 1300 582 594 1220 1260
Urinary protein, mg/L 493 910 532 541 880 890
Mean (SD):
MDA, nmol/g wet tissue 48.3 (9.6) 96.8 (11.7) 53.6 (10.2) 49.6 (8.9) 92.4 (9.4) 101.4 (8.1)
NOx, nmol/g wet tissue 41.2 (10.3) 84.4 (10.6) 46.2 (11.2) 42.3 (9.6) 78.2 (6.5) 74 (8.5)
GSH, µmol/g wet tissue 1.8 (0.2) 1 (0.3) 1.6 (0.2) 1.8 (0.2) 1.1 (0.2) 1.1 (0.3)
Semi-quantitative analysis of renal histology
Degree of necrosis
0 8 – – – – –
1 – – 5 4 – –
2 – – 3 4 – –
3 – 4 – – 4 5
4 4 – – 4 3
Histopathology: immunohistochemical staining score
iNOS
0 – – – – – –
1 – – – – – –
2 4 – – – – –
3 4 – 5 4 – –
4 – – 3 4 – –
5 – 3 – – 4 3
6 – 3 – – 2 3
7 – 2 – – 2 2
p65
0 – – – – – –
1 – – – – – –
2 3 – – – – –
3 5 – 4 3 – –
4 – – 4 5 – –
5 – 3 – – 3 2
6 – 3 – – 2 3
7 – 2 – – 3 3

Group 1, control; group 2, gentamicin for 10 days; group 3, gentamicin plus PD; group 4, gentamicin plus SZ; group 5, gentamicin plus distilled water; group 6,
gentamicin plus ammonium hydroxide for 10 days.

expressed and was not inhibited by the
NF-κB inhibitors. In the present study,
immunohistochemically, iNOS and P65
expression was significantly less in rats given
gentamicin and NF-κB inhibitors (PD and SZ)
than in rats given gentamicin only.
Sakurai et al. [59] previously reported that
prophylactic PD in rats with immune-
mediated glomerulonephritis attenuated
proteinuria and NF-κB activation. The exact
mechanisms of the anti-proteinuric effect
and its relation to NF-κB inhibition in the
latter model are unknown, but it might be
related to the potent antioxidant properties of
PD [21].
We showed that administration of NF-κB
inhibitors (PD and SZ) caused significantly
lower urinary protein levels than in rats
treated with gentamicin alone and
gentamicin + NH3 (Table 1). Decreases
in GFR and increases in serum creatinine
are usually not apparent until 7–10 days of
aminoglycoside treatment [9,10,27]. In
the present study, we showed that in rats
given gentamicin (100 mg/kg/day) for
10 consecutive days there was no increase
in blood urea nitrogen or serum creatinine
level. It is known that proximal tubules are
affected by short-term gentamicin treatment
and there is initially no glomerular damage.
This clarifies why there was no marked change
in GFR and blood creatinine levels in rats
given gentamicin in the short-term
[9,10,27,60].

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 T U G C U ET AL.

FIG. 1. A, Normal tubular structure in the control, group 1 (H&E, × 400); B, disseminated necrosis in the cortical tubules in group 2; desquamation of the glandular
epithelium and necrosis in the form of granular eosinophilic material (immunohistochemical staining, ×400); C, necrosis in a few proximal tubules in group 3
(H&E, × 100).

A B C

FIG. 2. Immunohistochemical staining, showing: A, focal mild staining (score 2) with iNOS in group 1 (×400); B, diffuse, intensive NF-κB/p65 positivity (score 7) of the
cortical tubules of group 2 (×400); C, Diffuse moderate iNOS positivity (score 6) in group 6 (×400); D, Mild NF-κB/p65 positivity (score 3) in some tubules of group 3
(×400); E, Mild NF-κB/p65 positivity (score 4) in some tubules of group 4 (×400); F, Low NF-κB/p65 positivity (score 2) in proximal tubules of group 2 (×100); G, Low
iNOS positivity (score 2) in proximal tubules of group 1 (×400).

A B C

D E F

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 SELECTIVE NF-κB INHIBITORS PREVENT NEPHROTOXICITY OF GENTAMICIN
Light microscopy showed complete necrosis
due to gentamicin toxicity; there was only
grade 1 or 2 necrosis in rats given gentamicin
and NF-κB inhibitors. We think that this
undesirable effect is related to the dose
administered, such that higher doses of
NF-κB inhibitors would prevent grade 1–2
necrosis. The combination of NF-κB inhibitors
and gentamicin substantially reduced
hyperproteinuria and restored the levels of
markers of oxidative stress nephrotoxicity.
NF-κB inhibitors not only prevented
peroxidation, GSH-Px depletion and NOx
production, but also decreased lipoperoxide
levels to control values, whereas those of NOx
and GSH-Px were significantly higher. PD and
SZ given concomitantly with gentamicin
(groups 3 and 4) caused significantly lower
MDA levels in the kidney tissues than in
rats treated with gentamicin alone and
gentamicin + NH3. Consequently, we propose
that NF-κB inhibitors (PD and SZ) act in the
kidney as potent scavengers of free radicals,
and they prevent the toxic effects of
gentamicin, i.e. biochemical changes and
tubular necrosis accompanying the
nephropathy.
These results also reinforce the significant
role of free radicals in the pathogenesis of
nephropathy induced by gentamicin. NF-κB
inhibitors like PD and SZ are probably the
most potent and specific antioxidants for
hydroxyl radicals.
CONFLICT OF INTEREST
None declared.
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Correspondence: Volkan Tugcu, Gül D-5 Blok
D:35 Postal code: 34538 Bahçesehir, Istanbul,
Turkey.
e-mail: volkantugcu@yahoo.com
Abbreviations: ROS, reactive oxygen species;
NF-kB, nuclear factor κ-B; (i)NO(S),
(inducible) nitric oxide (synthase); PD,
pyrrolidine dithiocarbamate; SZ,
sulfasalazine; GSH-Px, glutathione
peroxidase; MDA, malondialdehyde; TBA,
thiobarbituric acid; BHT, butylated
hydroxytoluene; GT, glutamyl transpeptidase;
H&E, haematoxylin and eosin.
© 2006 THE AUTHORS
© 2006 BJU INTERNATIONAL