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ISSN: 0148-0545 (print), 1525-6014 (electronic)
Drug Chem Toxicol, Early Online: 18
! 2013 Informa Healthcare USA, Inc. DOI: 10.3109/01480545.2013.866137

Ameliorative effects of Bacopa monniera on lead-induced oxidative

stress in different regions of rat brain
Manoj Kumar Velaga1, Charan Kumar Basuri1, Kendra S. Robinson Taylor2, Prabhakara Rao Yallapragada1,
Sharada Rajanna2, and Bettaiya Rajanna2
Department of Zoology, Andhra University, Visakhapatnam 530003, India, and 2Department of Biological Sciences, Alcorn State University,
Lorman, Mississippi, USA

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Abstract

Keywords

Bacopa monniera is a rejuvenating herb for brain cells enhancing learning and cognitive ability.
In the present investigation, the ameliorative effects of Bacopa monniera were examined
against lead-induced oxidative stress in different regions of rat brain. Male rats were divided
into five groups: control (1000 ppm sodium acetate) and exposed (1000 ppm lead acetate) for
4 weeks; DMSA (Meso-2,3-Dimercaptosuccinic acid)-treated (90 mg/kg body weight/day);
Bacopa monniera-treated (BM) (10 mg/kg body weight/day) and a combination of BM DMSA
for seven consecutive days after 4 weeks of lead exposure. After treatment, the whole brain was
isolated by sacrificing rats and four regions were separated namely cerebellum, hippocampus,
frontal cortex and brain stem. Results indicated a significant (p50.05) increase in reactive
oxygen species (ROS), lipid peroxidation products (LPP) and total protein carbonyl content
(TPCC) in association with tissue metal content in all the four regions of brain for exposed
group compared with their respective controls. However, the lead-induced ROS, LPP, TPCC and
tissue metal content were lowered on treatment with Bacopa monniera, almost reaching the
control group values in all the above brain regions compared to DMSA and a combination
therapy. Results suggest that Bacopa monniera can mitigate the lead induced-oxidative stress
tissue specifically by pharmacologic interventions which encompass both chelation as well as
antioxidant functions.

Antioxidant activity, Bacopa monniera, brain,

lead toxicity, oxidative stress

Introduction
Lead is one of the most widely studied neurotoxicant. It
adversely affects neuronal system (Cheng et al., 2012). Lead
has been reported as a potent neurotoxicant and has
widespread distribution including natural and anthropogenic
sources, such as e-waste which is an emerging problem
in developing and developed countries (Chen et al., 2011;
Patra et al., 2011). Exposure to lead has been recognized as
a major public health risk, particularly in developing
countries (Flora et al., 2012). Recent investigations have
established a link between lead contamination and memory
impairment in humans (Sadiq et al., 2012; Wijngaarden et al.,
2009). Lead exposure causes an increase in reactive oxygen
species (Bondy & Guo, 1996; Patra et al., 2011) and alters
nitrite and nitrate levels in rat brain (Chen et al., 2000).
Increase in serotoninergic neurotransmission, anxiety behavior (Sansar et al., 2011) and disruption of calcium homeostasis were reported after lead intoxication (Rajanna et al.,
Address for correspondence: Dr. Y. Prabhakara Rao, Ph. D., Professor,
Division of Animal Physiology & Toxicology, Co-ordinator, MHIRT
Program (USA), Andhra University, Department of Zoology,
Visakhapatnam-530003, India. Tel: +91 98480 42829 (mobile): +91
(891) 2573184 (residence). Fax: +91 (891) 2525611/2755324. E-mail:
yprabhakararao@yahoo.com

History
Received 2 May 2013
Revised 16 September 2013
Accepted 11 November 2013
Published online 12 December 2013

1996; Singh & Jiang, 1997). Lead mimics calcium and may
interfere with calcium signaling pathways, normal synaptic
signaling events and can even inhibit calcium channels (Neal
& Guilarte, 2010). Lead also induces astroglial changes which
may inhibit neuronal function leading to altered behavioral
abnormalities in locomotor activity, exploratory behavior,
learning and memory (Pachauri et al., 2009; Sansar et al.,
2011). Electron microscopic studies by Zhang et al. (2009)
revealed mitochondrial swelling, disruption, cristae loss and
nissil body dissolution on lead exposure. Lead was known to
alter the levels of lipid peroxidation products, antioxidant
enzymes and the apoptotic proteins in different brain regions
tissue-specifically (Bokara et al., 2008; Kiran et al., 2009; Xu
et al., 2008). Even its effects on the glutathione system and its
related enzymes were attributed to differences in the tissue
lead and susceptibility to oxidative stress (Bokara et al.,
2009).
At present Meso-2, 3-Dimercaptosuccinic acid (DMSA) is
the only known chelator approved by US FDA for treating
lead toxicity. DMSA plays a pivotal role in mobilization of
lead and recovery from oxidative stress (Saxena et al., 2005).
BAL (British anti-Lewisite) and DMPS (2,3-Dimercapto-1propanesulfonic acid) are also used for recovery of Deltaamino levulinic acid dehydratase (-ALAD) activity in
lead-induced mice (Santos et al., 2006). Recovery from lead

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M. K. Velaga et al.

induced biochemical and immunological alteration following

DMSA and calcium EDTA administration in rats was reported
by Flora et al. (1998). Flora et al. (2003) further studied
the beneficial effect of antioxidants and thiol chelators in
chronically lead exposed rats. However, the combined
administration of a chelator (DMSA and its analogue) and
antioxidant (n-acetylcysteine) was more effective in combating acute toxicity induced by lead (Pande et al., 2001).
Bacopa monniera Linn, also called as Brahmi, is an annual
creeping herb found throughout India, China, and Taiwan and
in some parts of the sub-continent (Nepal, Sri Lanka). It is
also found in USA in wet, damp and marshy regions (Barrett
& Strother, 1984). Bacopa is recommended as a nerve tonic,
brain tonic and memory, cognition enhancer, astringent, antipyretic, anti-epileptic and anti-inflammatory agent (Russo &
Borrelli, 2005; Singh & Dhawan, 1992). Its ethanolic extract
was found to inhibit the amnesic effects induced by scopolamine, electroshock and immobilization stress (Singh &
Dhawan, 1997). Superoxide dismutase, catalase and glutathione peroxidase activity showed an increase in different brain
regions on administration of Bacopa monniera extract
(Bhattacharya et al., 2000). The plant extract possesses
active principles for curing CNS (Central nervous system)
ailments and also for depressant activity. In addition, the
isolated bacosides from this plant extract have been extensively studied in several laboratories for their nootropic action
and neuro-pharmacological effects (Ganzera et al., 2004).
Bacopa monniera was known to ameliorate aluminiuminduced neurotoxicity in the cerebral cortex of the rat brain
(Jyoti et al., 2007). This was used to modulate the antioxidant
responses in brain and kidney of diabetic rats (Kapoor et al.,
2009). Plant extracts rich in antioxidants like curcuminoids,
curcumin have been used as a potent source to ameliorate
lead-induced oxidative stress in the brain regions (Dairam
et al., 2007). From the above literature, it is clear that many
attempts have been made to study the ameliorate effects of
plants against metal toxicity. In the present investigation, an
attempt has been made to test the efficacy of aqueous plant
extract (Bacopa monniera) alone and a combinatorial therapy
with DMSA against lead-induced oxidative stress. However,
the present study is a preliminary investigation to examine the
effect of crude extract of Bacopa against lead toxicity. Further
studies need to be conducted with a purified molecule to
study its ameliorative effect on lead toxicity.

Methods
Chemicals
Lead acetate (99.8%), Thiobarbituric acid (TBA), 2, 7
Dichlorofluorescein diacetate (DCF-DA), Meso-2, 3Dimercaptosuccinic acid (DMSA) and Guanidine hydrochloride were purchased from Sigma Chemical Co. (St. Louis,
MO). All other chemicals were obtained from SISCO
Research Laboratories Private Limited (Mumbai, India).

Drug Chem Toxicol, Early Online: 18

(Chemiloids) (Brindavan colony, Vijayawada, Andhra

Pradesh, India). This was prepared by following the method
adopted by Rehni et al. (2007). The powder of Bacopa
monniera was suspended in distilled water and was administered orally. Analysis of Bacopa monniera powder showed
no traces of heavy metals (Data not given).
Animals and treatment
The male rats of Wistar strain (100120 g) were purchased
from Mahaveera Enterprises (Hyderabad, India) and they
were maintained in the animal facility for four days before
they were used for experimentation. They were given free
access to feed (Pranav Agro Industries, India) and water
ad libitum. Analysis of feed and water showed no traces of
lead and other heavy metals. The study was approved by the
Institutional Animal Ethical Committee (IAEC). A total of
40 rats were treated with lead acetate (1000 mg/L) (Neal
et al., 1998) through drinking water for a period of 4 weeks
and parallel controls (10 rats) were maintained on sodium
acetate (1000 ppm). Both the solutions were prepared daily
with distilled water. Group-I (10 rats) control received sodium
acetate; Group-II (10 rats) exposed to lead acetate; Group-III
(10 rats) received individually DMSA at a dose of 90 mg/kg
body weight/day (i.p) (Saxena et al., 2005) for seven
consecutive days after four weeks of lead exposure. GroupIV (10 rats) received the aquatic plant extract of Bacopa
monniera (10 mg/kg body weight/day) (Jyoti et al., 2007) by
oral intubation for seven consecutive days after four weeks of
lead exposure. Group-V (10 rats) received DMSA (90 mg/kg
body weight/day; i.p) plus aquatic plant extract of Bacopa
monniera (10 mg/kg body weight/day; oral) for seven consecutive days after four weeks of lead exposure. The
treatments and supply of food to rats were stopped six
hours before sacrifice. The rats belonging to control and
exposed were sacrificed after four weeks of lead exposure
whereas the Group-III to V were sacrificed after one week
treatment with plant extract (Bacopa monniera) and DMSA.
Sacrifice was done by cervical dislocation and the whole
brains were isolated immediately on ice. The brains were
washed in cold normal saline (0.85% NaCl) solution and
different regions namely cerebellum, hippocampus, frontal
cortex and brain stem were separated on ice. Because of the
small amount of tissue, the hippocampus was pooled separately for all the groups. Reactive oxygen species, lipid
peroxidation products, protein carbonyl content were determined to study the oxidative damage and its recovery after
Bacopa administration (even along with DMSA). Metal
content was determined separately and this was done to
study the tissue lead burden and to know the chelating
property of Bacopa monniera extract.
Biochemical assays
Protein concentration in samples was determined following
Lowry et al. (1951) using bovine serum albumin as standard.

Bacopa monniera aqueous extract

The aqueous extract of Bacopa monniera (Brahmi) was
obtained as a gift sample in the form of greenish brown color
dried powder (Product code C/LMN/BAMO-01, Batch
No. L10121221) from M/S. Laila Impex Private Limited

Reactive Oxygen Species (ROS)

ROS levels in the tissues were determined using the method
of Bondy & Guo (1996). Fresh tissue homogenate (10%)
was prepared in 0.32 M sucrose solution. The contents were

DOI: 10.3109/01480545.2013.866137

Ameliorative effects of Bacopa monniera on lead-induced oxidative stress

centrifuged at 1800  g for 10 min. Pellet was discarded and

the supernatant was centrifuged at 31 500  g for 10 min to
obtain the crude synaptosomal pellet (P2). The P2 pellet was
suspended in HEPES buffer (120 mM NaCl, 2.5 mM KCl,
0.1 mM MgCl2, 6.0 mM glucose, 1.0 mM CaCl2, 5.0 mM
NaHCO3, 10 mM HEPES, pH 7.4) to a concentration of 0.1 g
equivalent/ml. Diluted fractions were incubated with 5 mM
2,7 Dichlorofluorescein diacetate (DCF-DA) (added from the
stock solution of 0.5 mM in 10 % ethanol) at 37  C for 15 min.
The obtained fluorescence was read at excitation wavelength
of 488 nm and emission wavelength of 525 nm in a spectroflurometer (Systronics, 152). ROS levels were expressed as
nanomoles of DCF-DA oxidized/15 min/mg protein.

supernatant was discarded. The pellet was re-suspended in

500 ml of 10 mM 2,4-dinitrophenyl hydrazine (DNPH) in 2 M
HCl and allowed to stand at room temperature for 1 hr, with
vortexing every 10 min at 4  C and the pellet was washed
three times with ethanol: ethyl acetate mixture. After the final
wash, the pellets were re-suspended in 500 ml of 6 M
guanidine hydrochloride (pH 2.3). The contents were
incubated at 37  C for 15 min and were centrifuged at
10 000  g for 10 min at 4  C. Carbonyl content was measured
in spectrophotometer (Rayleigh UV- 9200) at 360 nm against
a reagent blank. The results were expressed as nanomoles of
carbonyl/ml tissue.
Metal estimation

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Lipid peroxidation products (LPP)

Tissue LPP levels were estimated following the spectrophotometric method of Ohkawa et al. (1979). Fresh tissue of each
brain region was used to prepare 10 % homogenate in 1.5 %
KCl. To 1 ml of the homogenate, 2.5 ml of TCA (Trichloro
acetic acid) was added to precipitate the proteins. The
contents were centrifuged at 2500  g for 10 min at 4  C. The
supernatant was discarded and the pellet was dissolved in
2.5 ml of 0.05 M H2SO4 and to this, 3 ml of 2 M thiobarbituric
acid (TBA) was added. Whole contents were incubated in
boiling water bath at 100  C for 30 min. The contents were
cooled and color was extracted into 4 ml of n-butanol. The
color was read at 530 nm using a spectrometer (Rayleigh
UV-9200) against the blank. The results were presented as
micromoles of MDA (Malondialdehyde) formed/g weight
of tissue.
Total protein carbonyl content (TPCC)
TPCC levels were quantified using a slightly modified
method of Levine et al. (1990). 10% homogenate was
prepared with fresh tissue in cold buffer (50 mM Phosphate
buffer containing 1 mM EDTA, pH 6.7). The homogenate
was centrifuged at 10 000  g for 15 min at 4  C and the
supernatant was precipitated with equal amounts of 20%
trichloroacetic acid and incubated on ice for 5 min. The tubes
were centrifuged at 10 000  g for 10 min at 4  C and the

Metal concentration was measured following the method of

Zachariadis et al. (1995). The analysis of metal content was
carried out with the wet tissue. A known quantity of the tissue
was kept in muffle furnace at a temperature of 600  C for
about 45 hrs to make into ash. The ash obtained was digested
with HNO3 and dissolved in a known amount of 0.01 N
HNO3. The final clear and colorless solution was used for
metal estimation with atomic absorption spectrophotometer
(Perkin Elmer A Analyst 200). Lead concentration was given
as microgram lead/gm wet weight of tissue.
Statistical analysis
The mean values (n 10) with standard deviations were
calculated. The significant differences between control,
exposed and treated groups were determined using one-way
ANOVA at p50.05 followed by Bonferronis multiple comparison test. One-way ANOVA was performed using Graph
Pad Prism version 5.0 for Windows, GraphPad Software (San
Diego, CA, www.graphpad.com).

Results
Effect of Bacopa monniera on ROS levels in different
brain regions
Figure 1 represents the data on reactive oxygen species
(ROS) in four regions of control, exposed, DMSA, BM and

Figure 1. Reactive Oxygen Species (ROS) in different brain regions of control, exposed and treated rats. Bars represent mean (n 10) and vertical lines
represent standard deviation. *Significantly different from their respective control groups at p50.05. #Significantly different from their respective
exposed groups at p50.05.

M. K. Velaga et al.

Drug Chem Toxicol, Early Online: 18

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BM DMSA treated groups of the rat brain. There was a

significant (p50.05) (Table 1) increase in the ROS activity
in all the regions of the exposed group compared with
their respective controls. All the treated groups i.e. DMSA,
BM and BM DMSA showed a region specific recovery in
all the four regions of brain. There was a significant
(p50.05) decrease in ROS activity in the hippocampus
region for BM and BM DMSA treated groups compared
to the control group (Table 1). The ROS levels restored
back to control in cerebellum and frontal cortex for
BM DMSA group. When compared to exposed, all the
treated groups showed significant (p50.05) decrease in
ROS levels (Table 2).
Effect of Bacopa monniera on LPP levels in different
brain regions
The results on lipid peroxidation products (LPP) are
presented in Figure 2 for four regions of control, exposed,
DMSA, BM and BM DMSA treated groups of the rat brain.
A significant (p50.05) (Table 1) increase was observed in
LPP values of all the brain regions of the exposed group
compared to their respective controls. Though all the treated
groups showed recovery, it was not uniform in all the four
regions of brain. The cerebellum and the hippocampus
recorded decrease in LPP for the BM treated group when
compared to their respective controls. Maximum recovery
was noticed in frontal cortex on combination therapy of
Bacopa monniera and DMSA. However, the LPP values of
brain stem were almost nearer to the control group for all the
three treated groups. LPP levels showed significant (p50.05)
decrease in all the treated groups when compared to exposed
(Table 2).

Table 1. Percent change over their respective control groups and levels
of significance in different brain regions of exposed, DMSA, BM and
BM DMSA groups.

Cerebellum Hippocampus
ROS
Exposed
106.2***
DMSA
7.1
BM
34.1***
BM DMSA
8.9
LPP
Exposed
28.0***
DMSA
17.8**
BM
12.5
BM DMSA
2.3
TPCC
Exposed
37.3***
DMSA
7.1
BM
18.1***
BM DMSA 40.6***
Metal
Exposed
95.5***
DMSA
4.1
BM
25.7***
BM DMSA
3.6

83.3***
16.0***
20.0***
36.5***

Frontal
cortex

Brain
stem

223.5*** 146.8***
52.0*** 67.7***
2.6
3.2
0.5
33.2**

53.1***
3.0
15.7**
7.4

40.8***
16.9***
10.6
5.3

38.6***
4.6
3.2
1.2

36.6***
8.1*
4.8
7.1

54.4***
0.5
14.7**
3.75

84.2***
15.1*
0.7
20.8**

183.4***
85.9***
73.8***
33.3***

52.7***
7.6
23.7***
2.7

419.0***
194.6***
324.4***
179.5***

indicates increase and indicates decrease.

* significant at 0.05% level. ** significant at 0.001% level.
*** significant at 0.0001% level.

Effect of Bacopa monniera on TPCC levels in different

brain regions
Figure 3 represents the data on total protein carbonyl content
(TPCC) in the four regions of control, exposed, DMSA, BM
and BM DMSA treated groups of the rat brain. The TPCC
values increased significantly (p50.05) (Table 1) in all the
four brain regions exposed to lead. However, there was a
significant (p50.05) decrease in TPCC in the cerebellum of
BM and BM DMSA groups, and only in BM treated group
of the frontal cortex when compared to their controls
indicating recovery. In the case of hippocampus all the
groups showed similar recovery and brain stem showed
highest recovery in the case of BM DMSA group. When
compared to exposed, all the treated groups exhibited
significant (p50.05) decrease in TPCC levels (Table 2).
Effect of Bacopa monniera on lead content in different
brain regions
Lead content in the brain regions was presented in Figure 4.
Lead content was significantly (p50.05) (Table 1) high in all
the brain regions of exposed group indicating tissue accumulation. Maximum chelation of lead was shown in cerebellum
of BM DMSA group followed by DMSA. However, BM
treatment resulted in only a partial chelation of the metal. In
the hippocampal region, there was decrease in metal content
of treated groups and the order of chelation was high in
BM DMSA group followed by BM and DMSA treated
groups. The frontal cortex recorded less metal content
showing the best recovery on combination therapy of
BM DMSA followed by DMSA and partial recovery was
seen in BM. The chelation of metal in brain stem was also
partial and the order of recovery was highest in combination
therapy of BM DMSA followed by DMSA and BM treated
groups. Metal concentration decreased significantly (p50.05)
in all the groups when compared to exposed group (Table 2).

Table 2. Percent change over their respective exposed groups and levels
of significance in different brain regions of DMSA, BM and
BM DMSA groups.

ROS
DMSA
BM
BM DMSA
LPP
DMSA
BM
BM DMSA
TPCC
DMSA
BM
BM DMSA
Metal
DMSA
BM
BM DMSA

Cerebellum

Hippocampus

Frontal
cortex

Brain
stem

48.0***
34.9***
47.1***

54.2***
56.3***
65.3***

53.3***
69.9***
69.2***

32.0***
58.1***
46.0***

35.7***
31.6***
23.7***

32.7***
44.9***
39.5***

17.0***
21.5***
25.3***

31.2***
25.5***
27.6***

32.3***
40.3***
56.7***

20.8***
22.5***
21.7***

34.9***
44.8***
37.7***

37.6***
46.2***
57.0***

46.7***
35.7***
50.7***

34.4***
38.7***
52.9***

29.5***
18.9***
32.8***

43.2***
18.3***
46.2***

indicates increase and indicates decrease.

* significant at 0.05% level. ** significant at 0.001% level.
*** significant at 0.0001% level.

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DOI: 10.3109/01480545.2013.866137

Ameliorative effects of Bacopa monniera on lead-induced oxidative stress

Figure 2. Lipid Peroxidation Products (LPP) in different brain regions of control, exposed and treated rats. Bars represent mean (n 10) and vertical
lines represent standard deviation. *Significantly different from their respective control groups at p50.05. #Significantly different from their respective
exposed groups at p50.05.

Figure 3. Total Protein Carbonyl Content (TPCC) in different brain regions of control, exposed and treated rats. Bars represent mean (n 10) and
vertical lines represent standard deviation. *Significantly different from their respective control groups at p50.05. #Significantly different from their
respective exposed groups at p50.05.

Figure 4. Metal content (Lead) in different brain regions of control, exposed and treated rats. Bars represent mean (n 10) and vertical lines represent
standard deviation. *Significantly different from their respective control groups at p50.05. #Significantly different from their respective exposed
groups at p50.05.

M. K. Velaga et al.

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Discussion
Lead is a potentially harmful and toxic metal, which
accelerates the imbalance between pro and antioxidants
levels. Lead causes severe pathologic effects and degenerative
changes to the vital organs of the body especially brain. Brain
is susceptible to oxidative damage due to low levels of
antioxidant enzymes and high concentration of unsaturated
fatty acids (Halliwell & Gutteridge, 1985). Increased oxidative damage in brain can lead to cognitive and sensory
impairment depending on the region of the brain susceptible
to damage. In the present study, results indicate a significant
(p50.05) increase in ROS, LPP and TPCC in all the four
brain regions exposed to lead in comparison to their
respective controls (Table 1). These results are in correlation
with tissue accumulation of lead in different brain regions of
exposed rats. Lead increases the formation of ROS (Bondy &
Guo, 1996). However, treatment with BM individually or
in combination with DMSA resulted in restoration of ROS,
LPP and TPCC almost reaching the control values in all the
above brain regions (Table 1). In correlation with these
changes, the tissue metal content also showed a decrease in
the above BM treated and combination therapy of BM
DMSA in comparison to exposed individuals. The overall
data indicate that lead-induced oxidative stress can be
ameliorated by administration of Bacopa monniera (BM)
individually or in combination with DMSA.
ROS are free radicals formed from molecular oxygen
during cellular respiration or aerobic metabolism. These
contain unpaired or odd number of electrons (Halliwell &
Gutteridge, 1989; Harman, 1992). Increase in ROS causes
oxidative damage to tissues by oxidizing lipids and proteins.
ROS are known to increase the oxidative damage directly and
indirectly by increasing the levels of LPP and these in turn
increase the concentration of protein carbonyl content thus
enhancing oxidative damage (Burcham & Kuhan, 1996).
Generally the levels of ROS in the cells are kept under control
by endogenous antioxidants and minimize the damage caused
by ROS. Bacopa monniera treated neurons expressed lower
level of reactive oxygen species, prolonged lifespan and
increase in antioxidant enzyme (SOD, CAT and GSH-PX)
activities (Bhattacharya et al., 2000; Limpeanchob et al.,
2008). Bacopa monniera showed a dose-dependent free
radical scavenging capacity and a protective effect on DNA
cleavage (Russo et al., 2003). In the present investigation ROS
showed a significant increase in exposed brain regions but
Bacopa monniera treatment resulted in reduction of ROS.
However the decrease in ROS might be due to the increased
activity of endogenous antioxidant enzymes after Bacopa
monniera treatment.
ROS formed during oxidative stress react with polyunsaturated fatty acids (present in cell membranes) to form lipid
peroxides which decompose and form malondialdehyde
(Lippman, 1983). Lipid damage was estimated by TBA-RS
(Thiobarbituric acid reactive species) assay. The increased
concentration of LPP indicated lead induced oxidative
damage. Increased lipid peroxidation can in turn increase
protein oxidation. Burcham & Kuhan (1996) reported a time
and concentration dependent increase in carbonyl contents of
proteins when incubated with toxic lipid peroxidation product

Drug Chem Toxicol, Early Online: 18

malondialdehyde. Antioxidant activity of Bacopa can be

speculated to that of increase in antioxidant enzymes
(Bhattacharya et al., 2000).
Treatment with Bacopa monniera extract significantly
increased the antioxidant enzymes such as CAT, SOD and the
levels of GSH, inhibited lipid peroxidation and reduced the
tumor markers (Rohini et al., 2004). Antioxidants are known
to delay or prevent the oxidation of cellular oxidizable
substrates (lipid and protein) (Mates, 2000). Nemiche et al.
(2007) reported a decrease in lipid peroxidation in rats treated
with a-tocopherol. Treatment with Bacoside-A significantly
restored towards normalization of the decreased levels of
vitamin-C and vitamin-E (Sumathi & Nongbri, 2008) which
are well known non-enzymatic antioxidants. Bacopa monniera extract exhibited both reducing and lipid peroxidation
inhibitory activities (Limpeanchob et al., 2008). In the present
study, LPP levels were significantly increased in the lead
exposed group and were restored back to control values
following Bacopa monniera treatment. The results of the
present investigation are in good correlation with the above
investigations suggesting the role of Bacopa monniera in
reducing the levels of LPP by increasing the activity of
endogenous antioxidants and also levels of non-enzymatic
antioxidants.
Oxidative damage to proteins leads to introduction of
carbonyl groups and this reduces the function of proteins
(Gibson et al., 1999; Stadman, 1992). Assessment of
oxidative damage to proteins is generally studied by
measuring the protein carbonyl content (Beal, 2002;
Berlett & Stadman, 1997; Chevion et al., 2000; Shacter,
2000). Treatment with Bacoside-A resulted in restoration of
non-enzymatic antioxidants such as vitamin-C and vitamin-E
(Sumathi & Nongbri, 2008). However, Bacopa monniera
administration showed an increase in antioxidant enzymes,
protein synthesis and protein activity in brain cells
(Bhattacharya et al., 2000). Non enzymatic antioxidants
such as vitamin-C, vitamin-E (Nemiche et al., 2007) and
antioxidant enzymes (Pocernich et al., 2000) are known to
decrease lipid peroxidation and protein carbonylation (oxidation). Limpeanchob et al. (2008) reported that treating
patients with Bacopa monniera extract may be an alternative
direction for ameliorating neurodegenerative disorders associated with the overwhelming oxidative stress as well as
Alzheimers disease. It is evident from the present investigation that oxidative damage occurs to proteins with lead
administration resulting increased total protein carbonyl
content (TPCC). However, significant (p50.05) reduction
in TPCC levels was found in the cerebellum after combination therapy followed by Bacopa monniera treatment.
Bacopa monniera prevented accumulation of lipid and
protein damage significantly, which resulted from aluminium
intake (Jyoti et al., 2007). These findings support that
Bacopa monniera is efficacious in protecting the brain from
oxidative damage as resulting from metal toxicity.
Similar results on neuro-protection by plant extracts have
been reported earlier. Supplementation of Centella asiatica in
aged rats was effective in reducing brain regional lipid
peroxidation products and protein carbonyl levels and in
increasing the antioxidant status (Subathra et al., 2005).
Polyphenols have been reported to exert their neuroprotective

DOI: 10.3109/01480545.2013.866137

Ameliorative effects of Bacopa monniera on lead-induced oxidative stress

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activity by enhancing memory, learning and cognitive function (Vanzour, 2012). In addition some of the plant products
like curcumin, quercetin, green tea catechins, balcalein and
luteolin are found to increase the neurotrophic factors
essential for good health of nervous system (Blaylock &
Maroon, 2012). Allium sativum also could give protection
against metal toxicity by chelating mercury, cadmium and
lead (Nwokocha et al., 2012). Li et al. (2013) studied
the protective effect of Mangiferin (a xanthone from
Mangifera indica) against subchronic developmental lead
toxicity in rats.
Metal chelation property of Bacopa monniera was found to
be partial in the present study. The current study obviously
shows recovery from the oxidative damage with decrease in
metal content.

Conclusion
Overall these results suggest that Bacopa monniera can
mitigate the lead induced oxidative stress tissue specifically
by pharmacologic interventions which encompass both chelation as well as antioxidant functions. From the above results
and discussion it can also thus be concluded that administration of aqueous extract of Bacopa monniera has many
beneficial effects in protecting the neurons from oxidative
stress caused by lead toxicity. This is justified based on the
biochemical parameters selected in the study which are good
indicators of oxidative stress. Bacopa monniera plays protective role against lead-induced oxidative impairment in
brain tissue. The exact pathways utilized by Bacopa monniera
to exhibit its neuroprotective role against lead-induced
neurotoxicity are not completely known and require more
studies in this context. Future studies also include dosedependent changes of Bacopa extract on lead-induced oxidative stress in rat brain.

Acknowledgements
The authors thank the administration of Andhra University for
providing research facilities for this international collaboration of undergraduate student research training program.

Declaration of interest
The authors declare that there is no conflict of interest. This
work was supported by NIH/NCMHD/MHIRT, Grant #9
T37MD001532 awarded to Bettaiya Rajanna at Alcorn State
University, Mississippi, USA. Kendra S. Robinson Taylor was
MHIRT (Minority Health International Research Training)
undergraduate student participant.

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