Food and Chemical Toxicology 47 (2009) 2224–2229

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Food and Chemical Toxicology

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Preventive effect of Tinospora cordifolia against high-fructose diet-induced

insulin resistance and oxidative stress in male Wistar rats
Singareddy Sreenivasa Reddy a, Pasurla Ramatholisamma b, Rasineni Karuna a, Desireddy Saralakumari a,*
a
Department of Biochemistry, Sri Krishnadevaraya University, Anantapur 515 003, Andhra Pradesh, India
b
Department of Biochemistry, Sri Venkateswara University, Tirupati, Andhra Pradesh, India

a r t i c l e i n f o a b s t r a c t

Article history: High intake of dietary fructose exerts a number of adverse metabolic effects. The aim of the present study
Received 6 February 2009 was to investigate whether aqueous extract of Tinospora cordifolia stem (TCAE) alleviates high-fructose
Accepted 5 June 2009 diet-induced insulin resistance and oxidative stress in rats. High-fructose diet (66% of fructose) and TCAE
(400 mg/kg/day) were given simultaneously for a period of 60 days. Fructose fed rats showed hypergly-
cemia, hyperinsulinemia, hypertriglyceridemia, impaired glucose tolerance and impaired insulin sensi-
Keywords: tivity (P < 0.05). TCAE treatment prevented the rise in glucose levels by 21.3%, insulin by 51.5%,
Diabetes mellitus
triglycerides by 54.12% and glucose–insulin index by 59.8% of the fructose fed rats. Regarding liver anti-
Oral glucose tolerance test
Medicinal plants
oxidant status, fructose fed rats showed higher values of lipid peroxidation (91.3%), protein carbonyl
Antioxidants groups (44%) and lowered GSH levels (42.1%) and, lowered activities of enzymatic antioxidants, while
Sweetener TCAE treatment prevented all these observed abnormalities. In conclusion, our data indicate the preven-
Metabolic syndrome tive role of T. cordifolia against fructose-induced insulin resistance and oxidative stress; hence this plant
could be used as an adjuvant therapy for the prevention and/or management of chronic diseases charac-
terized by hyperinsulinemia, hypertriglyceridemia, insulin resistance and aggravated antioxidant status.
Ó 2009 Elsevier Ltd. All rights reserved.

1. Introduction
The last 25 years have witnessed a marked increase in total per
capita fructose intake as a sweetener in the food industry, primar-
ily in the form of sucrose (a disaccharide consisting of 50% fruc-
tose) and high-fructose corn syrup (HFCS; 55–90% fructose
content) (Bray et al., 2004). Processed-food manufacturers often
prefer HFCS to sucrose because it is inexpensive, sweeter and
mixes well in many foods. The increase in HFCS consumption far
exceeds the increases in intake of any other food or food group.
The disturbing fact is fructose consumption (excluding that which
occurs naturally in fruits and vegetables) increased from less than
0.5 g/day in 1970 to more than 40 g/day in 1997 (more than an
80-fold increase) (Gaby, 2005).
Concern has arisen because of the realization that fructose, at
elevated concentrations, can promote metabolic changes that are
actually or potentially deleterious, e.g., hyperlipidemia, hyperinsu-
linemia, insulin resistance, hyperuricemia, hypertension, glucose
intolerance and non-enzymatic fructosylation of proteins (Thor-
Abbreviations: AUC, area under the curve; CAT, catalase; GPx, glutathione
peroxidase; GR, glutathione reductase; GSH, reduced glutathione; GST, glutathione-
S-transferase; HFCS, high-fructose corn syrup; OGTT, oral glucose tolerance test;
ROS, reactive oxygen species; SOD, superoxide dismutase; TC, Tinospora cordifolia;
TCAE, aqueous extract of Tinospora cordifolia stem.
* Corresponding author. Tel.: +91 08554 255879; fax: +91 08554 255805.
E-mail address: skumari1@yahoo.co.in (D. Saralakumari).
burn et al., 1989; Hwang et al., 1987; Reddy et al., 2008; Dills,
1993). In addition, excessive fructose consumption may be respon-
sible in part for the increasing prevalence of obesity, diabetes mel-
litus, non-alcoholic fatty liver disease and cardiovascular diseases
(Jurgens et al., 2005; Reaven, 1988; Reiser, 1985).
Rats fed with a high-fructose diet form a model of diet-induced
insulin resistance, associated with hyperinsulinemia, hypertriglycer-
idemia and glucose intolerance (Thorburn et al., 1989). Recently, anti-
oxidants are found to be effective in preventing a majority of the
abnormalities induced by high-fructose diet (Faure et al., 1997, 1999).
Tinospora cordifolia (Menispermaceae) is a glabrous, succulent,
climbing shrub distributed throughout tropical Indian subcontinent.
The plant is commonly known as Guduchi, Giloy or Amritha. This
plant has been widely used in the Indian System of Medicine (Ayurv-
eda) as Rasayana for the treatment of diabetes, jaundice, rheumatoid
arthritis, gout, general weakness, skin diseases and infections (Deva-
sagayam and Sainis, 2002). It is known to have hepatoprotective
(Nagarkatti et al., 1994), immunostimulatory (Kapil and Sharma,
1997) and hyperlipidemic properties (Prince and Menon, 1999). A
variety of constituents have been isolated from this plant belong
to different classes such as alkaloids, diterpenoid lactones, glyco-
sides, steroids, sesquiterpenoid, phenolics, aliphatic compounds
and polysaccharides (Singh et al., 2003). Although, the antidiabetic
and antioxidant activities of this plant in experimentally diabetic
rats has been well documented in scientific literature (Prince and
Menon, 1999; Grover et al., 2001), studies regarding its efficacy in

0278-6915/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.fct.2009.06.008
 S.S. Reddy et al. / Food and Chemical Toxicology 47 (2009) 2224–2229
preventing insulin resistance which plays a role in pathophysiology
of type 2 diabetes mellitus have not been undertaken.
The anti-hyperglycemic, anti-hyperlipidemic and antioxidant
properties of T. cordifolia (TC) (in diabetic rat model) prompted
us to design the present study to investigate whether management
with TC has any preventive effect on plasma glucose, insulin, tri-
glycerides, hepatic lipid peroxidation and activities of both enzy-
matic and non-enzymatic antioxidant status in fructose fed rat
model of insulin resistance.
2. Materials and methods
2.1. Chemicals
Thiobarbituric acid and pyrogallol were obtained from the Sigma Chemical Co.,
St. Louis, MO, USA. All other chemicals and solvents were of analytical grade and
were from Sisco Research Laboratories (P) Ltd., Mumbai, India.
2.2. Plant extract
An aqueous extract of T. cordifolia stem (TCAE; brown, dry powder with Lot No.
L5111031) was procured from the manufacturers and exporters of herbal extracts,
Ms. Plantex Pvt. Ltd., Vijayawada, Andhra Pradesh, India. Procedure followed by the
firm for the preparation of extract is as follows: The plant was identified by Dr. K.
Narasimha Reddy, Taxonomist, Laila Impex R&D Center, Vijayawada. The collected
plant sample (stem) was washed thoroughly with tap water, dried at room temper-
ature away from sun light, cut into small pieces and then powdered. The aqueous
extract was prepared by cold maceration of stem powder in drinking water for
7 days. The extract was filtered, concentrated under reduced pressure and finally
dried in a vacuum desiccator. Herb-to-product ratio was 10:1. A voucher specimen
has been deposited in the Department of Biochemistry, Sri Krishnadevaraya Univer-
sity, Anantapur, under number SK-TC-08.
2.3. Control and fructose diet
The control diet for the rats contained 66% starch, 15% protein, 8% fat, 4% cellu-
lose, 1% of each mineral and vitamin mix. The fructose diet contained 66% of fruc-
tose instead of starch and remaining composition is same as that of the control diet.
Both the diets were obtained from National Centre for Laboratory Animal Sciences,
National Institute of Nutrition (Hyderabad, India).
2.4. Animals
Male Albino Wistar rats (140–160 g) used for the present study were procured
from Sri Venkateswara Enterprises (Bangalore, India). The animals were acclima-
tized for 7 days in our animal house (Regd. No. 470/01/a/CPCSEA) before dietary
manipulation. They were housed two per cage in an air-conditioned room
(22 ± 2 °C) with 12 h light/dark cycle and had free access to standard pellet diet
and water. All the procedures were performed in accordance with the Institutional
Animal Ethics Committee.
2.5. Experimental design
All the animals were 6 weeks of age, weighing around 200 g at the time of die-
tary manipulation. Animals were randomly assigned into four groups of eight each
as given below:
 Group-C: normal control rats, received tap water and control diet,
 Group-F: fructose fed rats, received tap water and fructose diet,
 Group-F+TC: TCAE treated fructose fed rats, received TCAE (400 mg/kg/day) and
fructose diet,
 Group-C+TC: TCAE treated normal rats, received TCAE (400 mg/kg/day) and con-
trol diet.
Vehicle (tap water for group-C and -F) and TCAE (dissolved in tap water) were
administered orally by gastric intubation. The animals were maintained in their
respective groups for 60 days. The dose of TCAE used in the current study was based
on the earlier report on the anti-hyperglycemic effect of this plant in experimental
diabetic rats (Grover et al., 2000) and our previous dose fixation studies for anti-
hyperglycemic effect of TCAE in alloxan-induced diabetic rats (data not shown).
The body weight, fasting plasma glucose, insulin and triglycerides of all animals
were measured on initial, 15, 30, 45 and 60th day of experiment.
2.6. Oral glucose tolerance test (OGTT)
At the end of experimental period (60 days), the 12-h fasted animals were sub-
jected to oral glucose tolerance test. For this, a 40% glucose solution was introduced
2225
directly into the stomach through a fine gastric catheter at a dose of 2 g/kg body
weight to conscious rats. Plasma glucose and insulin levels were determined at 0
(before glucose administration), 30, 60 and 120 min after glucose administration.
2.7. Measurement of glucose–insulin index
The action of insulin on glucose disposal rate was measured using the glucose–
insulin index, which is the product of the areas under the curve (AUC) of glucose
and insulin during the glucose tolerance test.
2.8. Sample collection
Blood was collected from 12-h fasted rats with capillary tube from retino-orbi-
tal plexus of the animals in fresh vials containing EDTA (10 mg/ml) as anticoagu-
lant. The samples were centrifuged at 3000 rpm for 5 min (MSE Micro Centaur,
UK) and the plasma obtained was aliquoted and frozen for insulin assay. Plasma
glucose and triglycerides were determined immediately. The blood sample col-
lected during oral glucose tolerance test was from the tail vein of animals.
After the experimental period the animals were fasted overnight and killed by
cervical decapitation. The body was cut open and liver was dissected out into ice-
cold saline and then thoroughly rinsed.
2.9. Biochemical measurements
The concentration of plasma glucose was measured by the glucose oxidase
method, using Span Diagnostic Kit (Surath, India). Plasma triglyceride level was
estimated by GPO–POD enzymatic method using the Monozyme Diagnostic kit
(Secunderabad, India). Insulin was determined by radioimmunoassay kit (RIAK-1)
provided by Bhabha Atomic Research Center (Mumbai, India) according to the
method of Yalow and Berson (1961). Human insulin was used for the preparation
of standard curve of insulin.
The concentration of lipid peroxidation intermediates, liver thiobarbituric acid
reactive substances (TBARS) was measured by following the method of Utley et al.
(1967), using 10% liver homogenate in 0.15 M KCl and expressed as nmol MDA
formed/15 min/mg protein. The extent of protein carbonyl groups (Levine et al.,
1990) and reduced glutathione (GSH) levels (Ellman, 1959) in liver were deter-
mined. Protein content in the liver homogenate was measured by the method of
Lowry et al. (1951).
2.10. Enzyme assays
Ten percent liver homogenate was prepared in ice-cold 0.15 M KCl, centrifuged
at 12,000 rpm for 45 min in Sigma Laboratory centrifuge 3K18 model, rotor No.
12150. The clear supernatant thus obtained was used for the assay of superoxide
dismutase (SOD; E.C.1.15.1.1; Soon and Tan, 2002), catalase (CAT; E.C.1.11.1.6;
Beers and Sizer, 1952), glutathione peroxidase (GPx; E.C.1.15.1.9; Rotsruck et al.,
1973), glutathione-S-transferase (GST; E.C.2.5.1.14; Habig et al., 1974) and glutathi-
one reductase (GR; E.C.1.8.1.7; Pinto and Bartley, 1969).
2.11. Statistical analysis
All results were expressed as means ± SEM for the number, n = 8 of animals in
the group as indicated in the figures and table. To determine the statistical signifi-
cance of clinical and laboratory findings Duncan Multiple Range test (DMRT) was
used. P values of less than 0.05 were regarded as significant.
3. Results
3.1. Effect of TCAE on body weight
The body weights of four groups of animals during experimen-
tal period are represented in Fig. 1A. No significant variation in
body weights of groups C+TC and F+TC was observed when com-
pared with group-C. Whereas, group-F also showed no significant
variation up to 15 days but a significant (P < 0.05) increase was
observed from 30 days onwards till the end of experimental period
when compared with group-C.
3.2. Fasting plasma glucose
There was no significant variation in the plasma glucose con-
centrations of group-C and C+TC throughout the experimental per-
iod (Fig. 1B). Group-F showed a gradual and significant increase in
plasma glucose levels from 30 days onwards till the end of exper-
2226 S.S. Reddy et al. / Food and Chemical Toxicology 47 (2009) 2224–2229

Fig. 1. Effect of TCAE on body weight, plasma glucose, insulin and triglycerides. Changes in body weight (A), fasting plasma glucose (B), fasting plasma insulin (C) and fasting
plasma triglycerides (D) of rats during the experimental period in group-C (j normal controls), group-F (d fructose fed rats), group-F+TC (N fructose fed rats treated with
TCAE at a dose of 400 mg/kg/day) and group-C+TC (. control diet fed rats treated with TCAE at a dose of 400 mg/kg/day). Values are means ± SEM. *Significantly different
(P < 0.05) from group-C. #Significantly different (P < 0.05) from group-F. 1Significantly different (P < 0.05) from group-C+TC. TCAE, aqueous extract of Tinospora cordifolia
stem.

imental period. The plasma glucose concentrations of group-F+TC
showed no significant variation throughout the experimental per-
iod except at 30 days.
3.3. Fasting plasma insulin
Group-C+TC showed statistically lower insulin levels at 45 and
60 days of experimental period when compared with group-C
(Fig. 1C). Group-F showed a gradual increase in plasma insulin dur-
ing the experimental period, showing significance at 30, 45 and
60 days with 1.4, 2.12 and 2.78-fold, respectively as compared to
group-C. The plasma insulin levels of group-F+TC were
45.52 ± 4.58, 64.12 ± 5.66 and 67.66 ± 6.34 l I.U/ml at 30, 45 and
60 days, respectively with 26%, 35% and 51% lower when compared
with group-F. The insulin levels of group-F+TC at the end of exper-
imental period were significantly lower than group-F but still sig-
nificantly higher than group-C.
3.4. Fasting plasma triglycerides
There was no significant change in plasma triglycerides
(Fig. 1D) of group-F up to 15 days, but from then there was a sig-
nificant increase till the end of experimental period. There was
28%, 123% and 133% increase in triglyceride levels in group-F at
30, 45 and 60 days, respectively when compared with group-C.
The triglyceride levels of group-F+TC showed no significant varia-
tion except at 45 days with 25% increase when compared with
group-C whereas significant lower triglyceride levels at 30, 45
and 60 days were obtained in group-F+TC compared with group-F.
3.5. Glucose tolerance and insulin sensitivity
The analysis of the glucose tolerance test and the comparison
between areas under the curve of glycemia during 120 min from
control and experimental groups show that fructose fed rats
 S.S. Reddy et al. / Food and Chemical Toxicology 47 (2009) 2224–2229 2227

developed glucose intolerance (Fig. 2A). The AUC of glucose during

OGTT of group-F was elevated to 1.3-fold when compared with
group-C. The AUC of glucose in group-F+TC was elevated by only
2% when compared with group-C. The AUC of glucose in group-
C+TC was significantly lower than group-C showing improved glu-
cose tolerance.
The AUC values of insulin during OGTT of group-F doubled the
AUC values of group-C. The AUC of insulin for group-F+TC were
lower by 96% compared to group-F and increased by only 4% when
compared with group-C (Fig. 2B).
The glucose–insulin index of four groups is represented in
Fig. 2C. The glucose–insulin index of groups C, C+TC and F+TC
has no significant difference. But glucose–insulin index of group-
F is significantly greater (166%) than group-C whereas group-
F+TC showed significantly lower by 60% when compared with
group-F.

3.6. Hepatic oxidative stress markers and antioxidants

Table 1 summarizes the levels of MDA, GSH, protein carbonyl

groups and activities of enzymatic antioxidants SOD, CAT, GST,
GPx and GR in the liver of control and experimental animals.
Group-F showed significantly higher levels of TBARS (91%) and
protein carbonyl groups (44%) as compared to group-C rats.
Group-F+TC showed significantly lower TBARS (34%) and protein
carbonyl groups (28%) when compared with group-F but still
significantly higher than group-C (27% and 4%, respectively).
Group-F showed depleted hepatic GSH levels (42%) as compared
to control, but treatment with TCAE limited the depletion to only
7%. Group-C+TC showed 1.5% increase in GSH levels when com-
pared to group-C rats.
The activities of enzymatic antioxidants SOD, CAT, GST, GPx and
GR were significantly lower (12%, 14%, 17%, 19% and 27%, respec-
tively) in group-F rats than in group-C rats. In group-F+TC, the
activities were significantly higher (15%, 13%, 16%, 20% and 23%,
respectively) as compared to group-F.

4. Discussion

Insulin resistance as a widespread feature of atherogenic dis-

eases predisposes the affected individuals to various diseases
including hypertension, dislipidemia, obesity, cardiovascular dis-
eases and type 2 diabetes mellitus (Zheng et al., 2005; Hotamisligil,
2000). Lowering endogenous insulin levels is a key step to success-
ful therapy directed at insulin resistance related diseases (Gold-
stein, 2002).
The development of insulin resistance in fructose fed rats is well
documented in the literature (Yagi et al., 1995; Thorburn et al.,
1989) and was also established in our laboratory (Reddy et al.,
2008). Results of the present study showed high-fructose feeding
in rats for 60 days leads to fasting hyperglycemia, hypertriglyceri-
demia, hyperinsulinemia, glucose intolerance and impaired antiox-
idant potential leading to the development of insulin resistance.
Further, as fructose-induced insulin-resistant animal model has
been recommended for assessing the therapeutic efficacy of insulin
sensitizers and drugs that are likely to have effect on insulin sensi-
tivity, we selected this model to study the efficacy of TCAE in pre-
venting insulin resistance. Fig. 2. Effect of TCAE on glucose tolerance and insulin sensitivity. Plasma
glucose (A) and plasma insulin (B) versus time during an oral glucose
The hyperglycemia, hypertriglyceridemia and hyperinsulinemia
tolerance test in group-C (j normal controls), group-F (d fructose fed rats),
observed in group-F at 30 days were further intensified by 45 and group-F+TC (N fructose fed rats treated with TCAE at a dose of 400 mg/kg/
60 days of fructose feeding. Hypertriglyceridemia after fructose day) and group-C+TC (. control diet fed rats treated with TCAE at a dose
feeding results from the enhanced rate of hepatic VLDL-triglyceride of 400 mg/kg/day). Insert area under the curve (AUC). Glucose–insulin index
synthesis (Thorburn et al., 1989; Zavaroni et al., 1982) and a (C). Values are means ± SEM. *Significantly different (P < 0.05) from group-C.
#
Significantly different (P < 0.05) from group-F. 1Significantly different
decrease in peripheral triglyceride clearance (Mayes, 1993). The (P < 0.05) from group-C+TC. TCAE, aqueous extract of Tinospora cordifolia
increased gene expression of several lipogenic enzymes, including stem.
2228 S.S. Reddy et al. / Food and Chemical Toxicology 47 (2009) 2224–2229
Table 1
Effect of TCAE on hepatic oxidative stress markers and antioxidants.
Parameter
TBARS (nmol of MDA/15 min/mg protein)
Reduced glutathione (lg/mg protein)
Superoxide dismutase (UA/mg protein)
Catalase (mmol of H2O2 decomposed/min/mg protein)
Glutathione-S-transferase (lmol of GSH–CDNB conjugate formed/min/mg protein)
Glutathione peroxidase (UB/mg protein)
Glutathione reductase (UC/mg protein)
Protein carbonyl groups (lmol/mg protein)
Values are given as mean ± SEM for eight rats in each group.
TCAE, aqueous extract of Tinospora cordifolia stem; MDA, Malondialdehyde; A, Amount of enzyme which gave 50% inhibition of pyrogallol autooxidation/min; B, lg of GSH
consumed/min; C, lmol of NADPH oxidized/min.
*
Significant as compared to group-C (P < 0.05 DMRT).
#
Significant as compared to group-F (P < 0.05 DMRT).
acetyl coenzyme-A carboxylase, fatty acid synthase and malic en-
zyme (Katsurada et al., 1990a,b) is also responsible for the en-
hanced synthesis of triglyceride in the liver of fructose fed rats.
Increased delivery of triglycerides to the muscle interferes with
the utilization of glucose, through the principles of Randle cycle
(Randle, 1998), impairing insulin action leading to hyperglycemia
and hyperinsulinemia. As insulin resistance and reduced insulin
binding have been reported in hypertriglycerolemic persons (Bie-
ger et al., 1984), this may be one mechanism by which fructose
diets promote insulin resistance. TCAE treatment completely pre-
vented fructose-induced hyperglycemia and partially prevented
hyperinsulinemia in group-F+TC. These positive effects can be
attributed to prevention of hypertriglyceridemia in these rats.
The plasma insulin levels of group-C+TC were significantly lower
from 30 days of treatment till the end of experimental period as
compared to group-C, without change in glucose levels indicating
the beneficial effect of TCAE in maintaining normoglycemia with
lower insulin levels.
The ability of insulin to stimulate glucose disposal is markedly
impaired as evidenced by increased glucose–insulin index in
group-F thus indicating decline in insulin sensitivity in peripheral
tissues associated with insulin resistance by fructose feeding. TCAE
treatment in fructose fed rats prevented glucose intolerance and
abnormal rise in glucose–insulin index indicating the ability of
TCAE in promoting the capability of insulin to stimulate glucose
disposal. However, the mechanism by which TCAE prevented insu-
lin resistance as induced by direct effect on target tissues or med-
iated by endogenous substances will be the subject of future
investigation.
The development of oxidative stress, an imbalance between
pro- and antioxidant status, has been shown to play an important
role in mediating insulin resistance, and therefore, we studied the
antioxidant potential. Decreased hepatic GSH levels, decreased
activities of antioxidant enzymes and increased lipid peroxidation
intermediates in group-F clearly indicates the development of oxi-
dative stress in these animals. Peroxidative deterioration of lipids
is evident from the increased levels of malondialdehyde, while
the increased protein carbonyl groups signify protein damage.
The increase in catabolism of fructose could be associated with
the cellular energy depletion that can increase the susceptibility
of cells to lipid peroxidation (Rajasekar et al., 2005). Furthermore,
it has been postulated that fructose can accelerate free radical pro-
duction similar to glucose. Reactive oxygen species (ROS) can
themselves reduce the activity of antioxidant enzymes such as
CAT and GPx (Datta et al., 2000). The decreased SOD activity in
fructose fed rats may be due to enhanced protein glycation by fruc-
tose as it is more reactive reducing sugar compared to others (glu-
cose and lactose) (McPherson et al., 1988; Oda et al., 1994). The
decreased GSH levels in group-F could be due to increased utiliza-
C F F+TC C+TC
* #
24.04 ± 1.89 45.99 ± 3.42 30.44 ± 2.66 22.61 ± 1.68
7.27 ± 0.36 4.21 ± 0.19* 6.78 ± 0.24# 7.38 ± 0.22
48.41 ± 1.34 42.45 ± 0.98* 49.72 ± 1.22# 47.9 ± 1.04
69.88 ± 1.92 60.01 ± 2.04* 68.92 ± 1.88# 69.95 ± 1.65
740.87 ± 12.56 609.75 ± 16.54* 722.5 ± 18.65# 744.25 ± 12.78
7.68 ± 0.24 6.24 ± 0.28* 7.79 ± 0.21# 7.84 ± 0.17
40.29 ± 0.84 29.30 ± 0.76* 38.22 ± 0.93# 41.43 ± 0.67
1.75 ± 0.08 2.52 ± 0.21* 1.82 ± 0.16# 1.72 ± 0.12
tion to trap free radicals, and/or decreased regeneration as evident
with the lower activity of glutathione reductase enzyme.
The antioxidant potential of TCAE against fructose diet-induced
oxidative stress is evident with lower levels of TBARS, higher GSH
levels and increased activities of antioxidant enzymes seen in
group-F+TC when compared with group-F. The antioxidant poten-
tial of this plant in alloxan-induced diabetic rats was well docu-
mented (Prince and Menon, 1999).
As oxidative stress has been suggested as one mechanism for
the detrimental effects of fructose, the antioxidant potential of
TCAE may be one among several mechanisms by which this plant
prevented insulin resistance. More recently, studies have linked
ROS production and oxidative stress to insulin resistance (Paolisso
and Giugliano, 1996; Ceriello, 2000). Through in vitro studies and
in animal models, it has been found that antioxidants improve
insulin sensitivity (Maddux et al., 2001; Rudich et al., 1999; Faure
et al., 1997). Several clinical trials, have also demonstrated that
treatment with vitamin E, vitamin C, or glutathione improves insu-
lin sensitivity in insulin-resistant individuals and/or patients with
type 2 diabetes (Evans and Goldfine, 2000; Jacob et al., 2000).
Further, it is important to confirm the safety of drug along with
its beneficial effects. T. cordifolia has been used for long periods in
Ayurveda, Siddha, Folkloric, Unani and other systems of medicine
for wide range of diseases without any evidence of adverse effects
or toxicity. In addition, studies conducted by Rege et al. (1999)
with the whole, aqueous, standardized extract of this plant did
not show any toxicity in both acute and sub acute toxicity studies.
5. Conclusion
Our data clearly indicate the beneficial effect of TCAE against
fructose-induced hyperglycemia, hyperinsulinemia, hypertriglyc-
eridemia and oxidative stress. These favorable effects might be
due to different types of active principles acting individually or
synergistically each with a single or a diverse range of biological
activities. Further studies are required to establish the mecha-
nism(s) underlying the defensive power of TCAE against fructose-
induced insulin resistance. The present study provides additional
evidence in support of use of TC in Ayurveda for diabetes mellitus
and other related metabolic disorders. In conclusion, aqueous
extract of T. cordifolia stem would seem useful as an adjuvant for
the prevention and/or management of pre-diabetic state of insulin
resistance and for subjects who need to increase insulin sensitivity.
Conflict of interest statement
The authors declare that there are no conflicts of interest.
 S.S. Reddy et al. / Food and Chemical Toxicology 47 (2009) 2224–2229
Acknowledgements
We appreciate National Centre for Laboratory Animal Sciences,
Hyderabad for the kind supply of fructose feed and control feed for
experimental animals. Thanks are also due to Dr. Appa Rao, Mr.
Vinay Kumar, Sri Venkateswara University, Tirupati for their help
in insulin assay.
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