Chemico-Biological Interactions 189 (2011) 100–106

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Chemico-Biological Interactions
journal homepage:www.elsevier.com/locate/chembioint

Ameliorative potential of S-allyl cysteine on oxidative stress in STZ

induced diabetic rats
Ganapathy Saravanan a,b,∗ , Ponnusamy Ponmurugan a,c
a Research and Development Centre, Bharathiyar University, Coimbatore, Tamil Nadu 641046, India
b Department of Biochemistry, Centre for Biological Science, K.S.R. College of Arts and Science, Thokkavadi, Tiruchengode, Tamil Nadu 637215, India c
Department of Biotechnology, K.S.R. College of Technology, Thokkavadi, Tiruchengode, Tamil Nadu 637215, India

article info abstract

Article history: Increased oxidative stress and impaired antioxidant defense mechanism are important factors in the pathogenesis and
Received 24 July 2010 progression of diabetes mellitus and other oxidant-related diseases. The present study was undertaken to evaluate the possible
Received in revised form 5 October 2010 protective effects of S-allyl cysteine (SAC) against oxidative stress in streptozotocin (STZ) induced diabetic rats. SAC was
Accepted 7 October 2010
administered orally for 45 days to control and STZ induced diabetic rats. The effects of SAC on glucose, plasma insulin,
Available online 14 October 2010
thiobarbituric acid reactive substances (TBARS), hydroperoxide, superoxide dismutase (SOD), catalase (CAT), glutathione
peroxidase (GPx), reduced glutathione (GSH), oxidized glutathione (GSSG) and GSH/GSSG ratio were studied. The lev-els
Keywords:
of glucose, TBARS, hydroperoxide, and GSSG were increased significantly whereas the levels of plasma insulin, reduced
Antioxidants
Diabetes mellitus glutathione, GSH/GSSG ratio, superoxide dismutase, catalase and GPx were decreased in STZ induced diabetic rats.
Streptozotocin Administration of SAC to diabetic rats showed a decrease in plasma glucose, TBARS, hydroperoxide and GSSG. In addition,
S-allyl cysteine the levels of plasma insulin, superoxide dismutase, cata-lase, GPx and reduced glutathione (GSH) were increased in SAC
treated diabetic rats. The above findings were supported by histological observations of the liver and kidney. The antioxidant
effect of SAC was compared with glyclazide, a well-known antioxidant and antihyperglycemic drug. The present study
indicates that the SAC possesses a significant favorable effect on antioxidant defense system in addition to its antidiabetic
effect.

© 2010 Elsevier Ireland Ltd. All rights reserved.

1. Introduction India and this is further set to rise to 69.9 million by the year 2025 [4].

Diabetes mellitus (DM) is the most significant chronic disease and cause
of death in modern society. Diabetes mellitus comprises a group of chronic
disorders characterized by hyperglycemia or diminished insulin secretion, or
both. DM involves high level of blood glucose, which contributes to an
increase in free radical production [1]. DM is a prevalent systemic disease
affecting a sig-nificant proportion of the population worldwide. The World
Health Organization (WHO) predicts that by the year 2025, 300 million
people will have diabetes mellitus [2]. Studies conducted in India in the last
decade have highlighted that not only in the prevalence of diabetes high but
also it is increasing rapidly in the urban pop-ulation [3]. The International
Diabetes Federation (IDF) estimates the total number of diabetic subjects to
be around 40.9 million in
Oxidative stress can be associated to an increased rate of reactive oxygen
species (ROS) generation, a decrease of antioxidant defense or a combination
of both. ROS-mediated alterations include damage to cells, tissues or organs
and are proposed as the major factors in the mechanism of several diseases
[1]. An increased production of oxygen-derived free radicals and the decrease
in the activity of free radical scavenger system have been reported in diabetes
[5]. It has also been proposed that an increase in oxidative stress could
contribute to tissue damage in diabetes [6]. Over the past decade there has
been an increased interest in oxidative stress and its role in the development of
complications of diabetes. Apart from the traditional antidiabetic treatment,
antioxidant therapy may benefit in diabetes.

In recent years, considerable focus has been given to an intensive search

for novel type of antioxidants present in plants for treating disease [7,8] and
some of the plants are used by the population as anti diabetic remedies [9].
Abbreviations: SAC, S-allyl cysteine; STZ, streptozotocin; kg, kilogram. Management of diabetes without any side effects is still a challenge to the
∗
Corresponding author at: Department of Biochemistry, Centre for Biological Science,
medical system. There is an increasing demand by patients to use the natural
K.S.R. College of Arts and Science, Thokkavadi, Tiruchengode, Tamil Nadu 637215, India.
Tel.: +91 9843954422.
products with antidiabetic activity, because insulin and oral hypoglycemic
E-mail address: saravana bioc@rediffmail.com (G. Saravanan).

0009-2797/$ – see front matter © 2010 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.cbi.2010.10.001
 G. Saravanan, P. Ponmurugan / Chemico-Biological Interactions 189 (2011) 100–106
drugs possess undesirable side effects [10]. Plants and its active compounds
provide useful sources for the development of drugs in the treatment of
diabetes mellitus. Clinical research has con-firmed the efficacy of several
plants and its active compounds in the modulation of oxidative stress
associated with diabetes mellitus [11].
S-allyl cysteine (SAC), a sulphur containing amino acid, derived from
garlic (Allium sativum Linn, Liliaceae), may constitute an alter-native to
insulin since both long- and short-term treatments with this compound correct
the hyperglycemia that occurs in diabetic model [12]. Treatment with SAC
also prevents the occurrence of various complications of diabetes in rats by
changing the blood glucose and protein metabolism [12]. SAC normalizes
hepatic car-bohydrate metabolism [13] and plasma and tissue glycoprotein
level [14] in diabetic rats. However, no studies have specifically addressed the
efficacy of SAC in STZ-induced oxidative stress in rats. Thus, the present
manuscript addresses the protective effect of SAC on lipid peroxidation,
antioxidant defense system and histopatho-logical changes in tissues of STZ-
induced diabetic rats. The effects produced by these treatments are compared
with standard drug glyclazide.
2. Materials and methods
2.1. Animals
Male Wistar rats of body weight 150–180 g were used for this study. The
animals were maintained in the animal house, Sas-tra University, Thanjavore,
India and fed on standard pellet diet (Amrut, Pune, India) and water ad
libitum. The protocol of this study was approved by Institutional Ethical
Committee of Sastra University.
2.2. Chemicals
SAC (99%) was purchased from LGC Prochem, Bangalore, India.
Streptozotocin was purchased from Himedia, Bangalore, India. All other
chemicals used were of analytical grade.
2.3. Induction of diabetes
Diabetes was induced [15] in overnight fasted adult Wistar strain albino
male rats weighing 150–180 g by a single intraperi-toneal injection of 55
mg/kg streptozotocin. Streptozotocin was dissolved in 0.1 M citrate buffer
(pH 4.5). Hyperglycemia was con-firmed by the elevated glucose levels
(above 250 mg/dl) in blood, determined at 72 h and then on day 7 after
injection.
2.4. Experimental design
The rats were divided into four groups each comprising a mini-mum of
six rats.
Group 1: normal control rats;
Group 2: diabetic control rats;
Group 3: STZ-treated rats given SAC (150 mg/kg body weight) orally using
an intragastric tube daily for a period of 45 days [12] and
Group 4: STZ-treated rats given glyclazide (5 mg/kg b.w. of rat) orally using
an intragastric tube daily for a period of 45 days [16].
After the last treatment (45 days), rats were fasted overnight and
sacrificed by cervical decapitation. Blood was collected and plasma was
obtained after centrifugation. It was used for the estimation of glucose. Liver
and kidney tissues were excised immediately from
101
the rats and stored in ice-cold containers. They were then homog-enized with
buffer, centrifuged and the supernatant was collected. Biochemical
estimations were carried out in the homogenates.
2.5. Biochemical estimations
2.5.1. Determination of blood glucose and plasma insulin
Plasma glucose was estimated colorimetrically by using com-mercial
diagnostic kits (Sigma Diagnostics (I) Pvt. Ltd., Baroda, India). Plasma
insulin assay was assayed by enzyme linked immunosorbent assay kit
(ELISA, Boerhriger Mannheim, Germany).
2.5.2. Determination of lipid peroxidation
The level of thiobarbituric acid reactive substances (TBARS) was
estimated by the method of Fraga et al. [17]. In brief, 0.5 ml of homogenate
was treated with 2 ml (1:1:1 ratio) TBA–TCA–HCl reagent (thiobarbituric
acid, 0.37%, 0.25 N HCl, 15% TCA) placed for 15 min in a water bath and
cooled. The absorbance of the clear supernatant was measured against
reference blank at 535 nm. Val-ues were expressed as mM/100 g-tissue.
The hydroperoxide was determined by the method of Jiang et al. [18].
Tissue homogenate (0.1 ml) was treated with 0.9 ml of Fox reagent (88 mg of
butylated hydroxy toluene (BHT), 7.6 mg of xylenol orange and 0.8 mg of
ammonium iron sulphate were added to 90 ml of methanol and 10 ml of 250
mM sulphuric acid) and incu-bated at 37 ◦ C for 30 min. Then the absorbance
was read at 560 nm. Hydroperoxides were expressed as mM/100 g-tissue.
2.5.3. Determination of reduced glutathione
Reduced glutathione was measured according to the method of Beutler
and Kelley [19]. The technique involved in protein precip-itation by
metaphosphoric acid and spectrophotometric assay at 412 nm of the yellow
derivative obtained by the reaction of super-natant with 5, 5 -dithio-bis-2-
nitrobenzoic acid (DTNB).
2.5.4. Determination of oxidized glutathione
Oxidized glutathione was measured according to the method described by
Aseni et al. [20] based on the principle of glutathione reductase enzyme
reducing GSSG to GSH with the concomitant oxidation of NADPH to
+
NADP . To 0.9 ml of 1.75 mol/l K3 PO4 buffer (pH 7.0) containing 20
mmol/l NEM was added 0.05 ml of sample extract and 0.025 ml of 10 mg/ml
of NADPH–Na solution. Absorbance at 340 nm was measured for 30 s
immediately after addition of 0.025 ml of (10 mg/ml) glutathione reductase
(GR) to the assay mixture.
2.5.5. Determination of superoxide dismutase
Superoxide dismutase (SOD) activity was assayed by the method of
Kakkar et al. [21]. 0.5 ml of tissue homogenate was diluted with 1 ml of
water. In this mixture, 2.5 ml of ethanol and 1.5 ml of chloroform (all reagents
chilled) were added and shaken for 1 min at 4 ◦ C then centrifuged. The
enzyme activity in the super-natant was determined. The assay mixture
contained 1.2 ml of sodium pyrophosphate buffer (0.025 M, pH 8.3), 0.1 ml of
186 m phenazine methosulphate (PMS), 0.3 ml of 30 m nitroblue tetra-zolium
salt (NBT), 0.2 ml of 780 m NADH, appropriately diluted enzyme preparation
and water in a total volume of 3 ml. Reaction was started by the addition of
NADH. After incubation at 30 ◦ C for 90 s the reaction was stopped by the
addition of 1 ml glacial acetic acid. The reaction mixture was stirred
vigorously and shaken with 4 ml of n-butanol. The intensity of the chromogen
in the butanol layer was measured at 560 nm against butanol blank. A system
devoid of enzyme served as control. One unit of the enzyme activ-ity is
defined as the enzyme reaction, which gave 50% inhibition of NBT reduction
in 1 min under the assay condition.
102 G. Saravanan, P. Ponmurugan / Chemico-Biological Interactions 189 (2011) 100–106

Table 1
Levels of lipid peroxides and hydroperoxides in liver of control and experimental groups of rats.

Groups TBARS Hydroperoxides

(mM/100 g) of (mM/100 g of
tissue tissue)

Control 8.84± 1.05 2.38 ± 0.84

Diabetic 14.26± 1.53a , * 4.32 ± 1.14a , *
Diabetic + SAC 9.93± 0.86b * 2.16 ± 0.95b , *
c* c,*
Diabetic + glyclazide 8.98± 1.3 2.47 ± 1.32
Values are mean ± SD, n = 6.
a Significantly different from control.
b Significantly different from diabetic control. c

Significantly different from diabetic control. * P <

0.001.
Fig. 1. Levels of blood glucose in control and experimental groups of rats. Values are mean ±
SEM, n = 6. a Significantly different from control. b Significantly different from diabetic
Table 2
control. c Significantly different from diabetic control. *P < 0.05. Levels of lipid peroxides and hydroperoxides in kidneys of control and experimental groups of
rats.
2.5.6. Determination of catalase
Groups TBARS Hydroperoxides
Catalase level was determined by Aebi’s modified colorimetric method (mM/100 g of (mM/100 g of
[22]. The reaction mixture (1.5 ml, vol.) contained 1.0 ml of 0.01 M tissue tissue)
phosphate buffer (pH 7.0), 0.1 ml of tissue homogenate and 0.4 ml of 2 M
Control 10.87 ± 1.53 2.83 ± 1.21
H2O2 . The reaction was stopped by the addition of 2.0 ml of dichromate– a
Diabetic 16.09 ± 1.47a , *** 4.71 ± 0.77a , **
b
acetic acid reagent (5% potassium dichromate and glacial acetic acid were Diabetic + SAC 9.69 ± 0.73b , *** 3.05 ± 0.72b , *
mixed in 1:3 ratio). Then the absorbance was read at 620 nm; CAT activity b c , *** c , **
Diabetic + glyclazide 10.33 ± 1.35 2.52 ± 0.75
was expressed as M of H2O2 consumed/min/mg protein. Values are mean ± SD, n = 6.
a Significantly different from control.
b Significantly different from diabetic control. c

Significantly different from diabetic control. * P <

2.5.7. Determination of glutathione peroxidase
GPx activity was measured by Paglia and Valentine’s method
[23] The reaction mixture contained 2.6 ml of 100 mmol/l phos-phate buffer
(pH 7.0) with 3 mmol/l EDTA, 0.05 ml of 10 mg/ml GSH solution, 0.1 ml
GR (10 mg/ml), 0.05 ml (10 mg/ml) NADPH–Na salt, 0.1 ml 90 mmol/l
hydrogen peroxide solution and 0.1 ml of sample. The GPx activity was
monitored by the decrease in absorbance due to the consumption of NADPH.
The absorbance was read at 340 nm.
2.6. Statistical analysis
All the results in tables were expressed as the mean ± SD for six animals
in each group and figure results were expressed in mean ± SEM. All the
grouped data were statistically evaluated with SPSS\10.0 software.
Hypothesis testing methods included one way analysis of variance (ANOVA)
followed by least significant differ-ence (LSD) test; significance levels at P <
0.01, 0.05 and 0.001 were considered to indicate statistical significance.
0.05.
**
P < 0.01.
***
P < 0.001.
Fig. 3 summarizes the concentration of GSSG and the level of GSH/GSSG
ratio in control and experimental rats. There was a significant (P < 0.05)
increased level of GSSG and concomitant decreased in the level of
GSH/GSSG in diabetic rats when compared to control rats. Oral treatment of
SAC and glyclazide tended to bring GSSG and GSH/GSSG ratio towards near
normal levels.
Tables 3 and 4 summarize the activities of superoxide dismutase (SOD),
catalase (CAT) and glutathione peroxidase (GPx) in the liver and kidneys of
control and experimental groups of rats. The activ-ities of SOD, CAT and
GPx in liver and kidneys were significantly (P < 0.05) lower in diabetic
control rats compared to diabetic rats. Treatment with SAC and glyclazide
showed a significant increase in activities of SOD, CAT and GPx in liver and
kidneys of diabetic rats.

Histopathological studies revealed normal central vein and radi-ating

hepatocytes in control liver (Fig. 4A) whereas diabetic control rats showed
3. Results
vascular congestion and mononuclear cellular infil-tration (Fig. 4B). SAC and
glyclazide treated rats revealed normal
Fig. 1 shows the levels of glucose and plasma insulin in control and
experimental animals. There was a significant (P < 0.05) eleva-tion in blood
glucose level with a significant decrease in plasma insulin levels in STZ
induced diabetic rats, compared with control rats. Administration of SAC and
glyclazide tended to bring blood glucose and plasma insulin towards near
normal levels.
Tables 1 and 2 show the concentration of TBARS, hydroperox-ides in
liver and kidneys of control and experimental groups of rats. The levels of
TBARS and hydroperoxides in diabetic rats were sig-nificantly (P < 0.05)
higher than control rats, whereas diabetic rats treated with SAC and
glyclazide restored the altered values to the near normal.

Fig. 2 shows the levels of reduced glutathione in liver and kid-neys of

control and experimental groups of rats. The decreased concentration (P <
0.05) of GSH was observed in diabetic rats. Administration of SAC and Fig. 2. Levels of GSH in liver and kidneys of control and experimental groups of rats. Values
glyclazide tends to bring the GSH level to near normal. are mean ± SEM, n = 6. a Significantly different from control. b Significantly dif-ferent from
diabetic control. c Significantly different from diabetic control. *P < 0.05.
 G. Saravanan, P. Ponmurugan / Chemico-Biological Interactions 189 (2011) 100–106 103

Table 3
Activities of superoxide dismutase, catalase and glutathione peroxidase in the liver of control
and experimental groups of rats.

Groups SOD CAT GPx

Control 7.78 ± 1.06 13.4 ± 0.72 7.10 ± 0.82

Diabetic 4.88 ± 1.08a , ** 9.33 ± 1.20a , *** 3.79 ± 1.17a , ***
Diabetic + SAC 6.82 ± 0.38b , * 11.37 ± 0.93b , * 5.54 ± 0.94b , *
c,* c , ** c , **
Diabetic + glyclazide 7.16 ± 1.57 12.28 ± 1.67 5.95 ± 0.99
Values are mean ± SD, n = 6.
Activity is expressed as: 50% of inhibition of epinephrine auto oxidation per min for SOD;
mole of hydrogen peroxide decomposed per min per mg of protein for catalase; mole of
glutathione oxidized per min per mg of protein for GPx.
a Significantly different from control.
Fig. 3. Level of GSSG and GSH/GSSG ratio in control and experimental rats. Values are mean
b Significantly different from diabetic control. c
± SEM, n = 6. a Significantly different from control. b Significantly different from diabetic
*
Significantly different from diabetic control. P < control. c Significantly different from diabetic control. *P < 0.05.
0.05.
**
P < 0.01.
***
P < 0.001. architecture with regenerating hepatocytes (Fig. 4C and D). Kid-ney of control
rat showed normal tubules and glomerulus (Fig. 5A), whereas kidney of
Table 4 diabetic control rat revealed intertubular haem-orrhage and mononuclear
Activities of superoxide dismutase, catalase and glutathione peroxidase in kidneys of control cellular infiltration (Fig. 5B). Kidney of SAC and glyclazide treated rats
and experimental groups of rats. revealed normal tubules and glomerulus (Fig. 5C and D).
Groups SOD CAT GPx

Control 10.15 ± 1.61 9.76 ± 0.55 7.54 ± 0.86

Diabetic 5.97 ± 1.47a , *** 4.38 ± 1.46a , *** 3.18 ± 0.90a , *** 4. Discussion
Diabetic + SAC 8.76 ± 0.96b , * 7.98 ± 0.83b , *** 5.56 ± 1.77b , *
Diabetic + glyclazide 9.24 ± 1.94
c , **
8.26 ± 2.01
c , ***
6.04 ± 0.77
c , ** Streptozotocin (STZ)-induced hyperglycemia in animals is con-sidered to
Values are mean ± SD, n = 6. be a good model for the preliminary screening of agents active against
Activity is expressed as: 50% of inhibition of epinephrine auto oxidation per min for SOD; diabetes [24]. In experiment with many animal species [25], streptozotocin
mole of hydrogen peroxide decomposed per min per mg of protein for catalase; mole of produces permanent diabetes that impersonates the pathological status found
glutathione oxidized per min per mg of protein for GPx.
a Significantly different from control. in human diabetes [26]. In the present study, the intraperitoneal administration
b Significantly different from diabetic control. c of strepto-zotocin effectively induced diabetes mellitus in rats. STZ-induced
Significantly different from diabetic control. * P < experimental diabetes may exhibit most of the diabetic compli-cations
0.05. mediated through oxidative stress [27]. During diabetic state, increased
**
P < 0.01. generation of ROS occur and cause an imbalance
***
P < 0.001.

Fig. 4. (A) Liver section of normal control showing normal central vein and radiating hepatocytes. H&E 400×. (B) Liver section of diabetic rats showing vascular congestion and mononuclear cellular
infiltration (arrow mark shows mononuclear cellular infiltration). H&E 400×. (C) Liver section from SAC treated animals revealing normal architecture with regenerating hepatocytes. H&E 100×. (D)
Liver section from glyclazide treated animals revealing normal architecture with regenerating hepatocytes. H&E 400×.
104 G. Saravanan, P. Ponmurugan / Chemico-Biological Interactions 189 (2011) 100–106
Fig. 5. (A) Kidney section from control showing normal tubules and glomerulus. H&E 400×. (B) Kidney section diabetic control revealing intertubular haemorrhage and mononuclear cellular
infiltration (arrow mark shows intertubular haemorrhage). H&E 400×. (C) Kidney section SAC treated animals revealing normal tubules and glomerulus. H&E 400×. (D) Kidney section glyclazide
treated animals revealing normal tubules and glomerulus. H&E 400×.
between the oxidant and antioxidant status [28]. Administration of STZ
resulted in a significant increase in the blood glucose level and reduction in
plasma insulin level. Sustained hyperglycemia has been identified as a
principle mediator of increased reactive oxy-gen species generation in
diabetes [29]. Oral administration of SAC decreased the blood glucose level
and increased plasma insulin level in diabetic rats. This may be due to the
insulin like effect of SAC on peripheral tissues, either by promoting glucose
uptake and metabolism, or by inhibiting hepatic gluconeogenesis [12].
Increased free radical observed in diabetic rats is attributed to chronic
hyperglycemia that damage antioxidant defense system [30]. Free radicals
may also be formed via the auto-oxidation of unsaturated lipids in plasma and
membrane lipids. The free rad-ical produced may react with polyunsaturated
fatty acids in cell membrane leading to lipid peroxidation. Lipid peroxidation
will in turn result in elevated production of free radicals [31]. Increased lipid
peroxidation impairs membrane functions by decreasing membrane fluidity
and changing the activity of membrane-bound enzymes and receptors [32]. Its
products are harmful to most of the cells in the body and are associated with a
variety of diseases [33]. Our present study showed a significant increase of
tissue thio-barbituric acid reactive substances (TBARS) and hydroperoxides
(HP) level in diabetic rats. The increased TBARS content of dia-betic rats
suggests that peroxidative injury may be involved in the development of
diabetic complications. TBARS and hydroperoxides levels in liver and
kidneys were significantly lower in the SAC and glyclazide-treated group
compared to the diabetic control rats. The above result suggests that the SAC
may exert antioxidant activities and protect the tissues from lipid
peroxidation.
GSH is a major endogenous antioxidant which counterbalance free radical
mediated damage. Studies have shown that the tissue GSH concentrations of
STZ-induced diabetic rats are significantly lower when compared with the
control rats [34]. It is well known that GSH is involved in the protection of
normal cell structure and function by maintaining the redox homeostasis,
quenching of free
radicals and participating in detoxification reactions. It is a direct scavenger of
free radicals as well as a co-substrate for peroxide detoxification by
glutathione peroxidases [35]. Loven et al. [36] had suggested that the decrease
in tissue GSH could be the result of decreased synthesis or increased
degradation of GSH by oxida-tive stress in diabetes. Increased oxidative
stress, resulting from a significant increase in aldehydic products of lipid
peroxidation has probably decreased the tissue GSH content [35]. In the
present study, the elevation of GSH levels in liver and kidneys was observed
in the SAC and glyclazide treated diabetic rats. This indicates that the SAC
and glyclazide can either increase the biosynthesis of GSH or reduce the
oxidative stress leading to less degradation of GSH, or have both effects.
GSH and GSSG levels are commonly used markers for oxida-tive stress.
The decrease in GSH content, which is shown in the results of the present
work, seems to be due to its oxidation to GSSG (oxidized glutathione) [37].
GSSG appears to be released from most cells as a consequence of oxidative
stress so that an oxida-tion of the cellular pool could shift the balance of GSH
and GSSG efflux and change the extra cellular redox state. This indicates that
GSH/GSSG redox system plays a significant role in diabetes. In other words,
a low GSH/GSSG ratio suggests increased oxidative stress. The GSH/GSSG
ratio seems to play a major role in the modulations of glucose homeostasis in
diabetes [38]. GSH infusion has been shown to increase GSH/GSSG ratio and
potentiate the insulin action in diabetic patients [33]. Thus, any drug that
replenishes glutathione may be able to reverse the oxidative damage caused in
diabetes mellitus and prevent the associated disorders. SAC, a known antiox-
idant probably strengthened the antioxidant status as evidenced by increase in
the GSH levels.
In diabetic state, insulin deficiency caused the impairment of glucose
utilisation leading to an increased generation of oxygen free radicals [35].
Studies have reported on the reduction of tissue SOD activities in STZ-
induced diabetic rats [39]. SOD is an important defense enzyme which
catalyses the dismutation of superoxide
 G. Saravanan, P. Ponmurugan / Chemico-Biological Interactions 189 (2011) 100–106
radicals to produce H2 O2 and molecular oxygen [40], hence dimin-ishing the
toxic effects caused by their radical. Wohaieb et al. [41] had suggested that
the reactive oxygen free radicals could inacti-vate and reduce the tissue SOD
activities. The observed decrease in SOD activity could result from
inactivation by H2 O2 or by gly-cation of enzymes [42]. Oral treatment of
SAC caused a significant increase in SOD activities in diabetic rats. This
means that the SAC can reduce reactive oxygen free radicals and improve the
activities of the tissue antioxidant enzymes.
Catalase (CAT) is a hemeprotein which catalyses the reduction of
hydrogen peroxides and protects the tissues from highly reac-tive hydroxyl
radicals [43]. The decrease in CAT activity could result from inactivation by
glycation of enzyme [44]. CAT reduces hydro-gen peroxide produced by
dismutation reaction. It also prevents the generation of hydroxyl radicals
thereby protecting the cellular constituents from oxidative damage in
peroxisomes. The reduced activity of CAT in STZ-treated rats results in the
accumulation of H2 O2 , which produces deleterious effects. In the present
study, it was observed that the SAC caused a significant increase in the activ-
ity of CAT in diabetic rats. This action is predominantly due to the
antioxidant nature of SAC and could involve a mechanism related to
scavenging activity.
Glutathione peroxidase (GPx), a selenium containing enzyme present in
significant concentrations, detoxifies H2 O2 to H2 O through the oxidation of
reduced glutathione [34]. GPx works together with glutathione in the
decomposition of H2 O2 or other organic hydroperoxides to non-toxic
products at the expense of reduced glutathione [35]. Reduced activities of
GPx may result from radical-induced inactivation and glycation of the
enzyme [34]. The low activity of GPx could be directly explained by the low
content of glutathione found in diabetic state, since glutathione is a substrate
and cofactor of GPx. Reduced activities of GPx in the liver and kidney have
been observed during diabetes and this may result in a number of deleterious
effects due to the accu-mulation of toxic products. In this context, other
workers also reported a decrease in the activity of GPx in the liver and kid-
neys of diabetic rats [45]. In the present study, administration of SAC and
glyclazide increased the activities of GPx in diabetic rats.
In the present study, the histopathological examinations of the liver and
kidneys of diabetic rats showed vascular conges-tion and mononuclear
cellular infiltration of the hepatocytes, as well as areas of intertubular
haemorrhage and mononuclear cellular infiltration. The reaction is provoked
by the increased production of highly reactive intermediates of STZ, which
are nor-mally detoxified by endogenous GSH [46]. The above pathological
changes were reduced in diabetic rats treated with SAC and gly-clazide.
In conclusion, the present investigation showed that SAC pos-sesses an
antioxidant activity and also protects lipid peroxidation and enhances its
effect on cellular antioxidant defense. This activity contributes to the
protection against oxidative damage in STZ-induced diabetes.
Conflict of interest statement
None.
Acknowledgements
The authors thank the management of K.S.R. College of Arts and Science,
Tiruchengode, India and Dr. N. Kannan, principal of this college for their
encouragement and also the management of Sastra University, Thanjavore,
India for providing facilities to do animal studies.
105
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