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41 ReferencesAnti-Bacterial and Anti-Oxidant Activity of Achyranthes Aspera Leaf Extract and Its Effect on Gall Bladder Stones
Abstract
In recent years, bacterial infections are increasing due to emergence of antibiotic resistance. Oxidative stress and gall bladder stones also pose a great health threat to millions. This study attempted the assessment of antibacterial, anti-oxidant and anti-gall bladder stones activity of an important medicinal plant Achyranthes aspera methanolic leaf extract. Anti-bacterial activity was performed by disc diffusion method. Significant susceptibility was observed against gram positive bacteria than gram negative strains. Anti-oxidant activity was studied in terms of 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging potential with ascorbic acid as standard. Highest DPPH scavenging activity for A. aspera and ascorbic acid was 59.21% and 92.41%, respectively. IC50 value was 472.93 µg/ml for A. aspera and 1.4965 µg/ml for ascorbic acid. Effects of A. aspera leaf extract on gall bladder stones were also investigated. 2.0 mg/ml extract concentration showed the highest amount of cholesterol release (39.69 mg/dl) and the highest amount of dry weight reduction (2.3 mg) from gall stones. Morphological changes like color change and fragility in gall stones were also observed at concentrations higher than 1.5 mg/ml.
Keywords: Medicinal Plant, Achyranthes aspera, Anti-bacterial, Anti-oxidant, Gall bladder stone, DPPH, IC50
Journal of Medicinal Plants Studies
Year: 2013, Volume: 1, Issue: 3
First page: (105) Last page: (117)
ISSN: 2320-3862
Online Available at www.plantsjournal.com
Journal of Medicinal Plants Studies
Vol. 1 Issue. 3 2013 www.plantsjournal.com Page | 105
Anti-Bacterial and Anti-Oxidant Activity of Achyranthes
Aspera Leaf Extract and Its Effect on Gall Bladder Stones
Md. Jakir Hossain1 , Laila Khaleda1, A.M. Masudul Azad Chowdhury1, Md. Arifuzzaman2, Mohammad Al-
Forkan*1
1. Department of Genetic Engineering and Biotechnology, University of Chittagong, Chittagong-4331,
Bangladesh.
[E-mail: alforkancu@hotmail.com; Tele: +88-01819383213]
2. Department of Biochemistry and Biotechnology, University of Science and Technology, Chittagong
(USTC), Foy’s Lake, Chittagong-4202, Bangladesh.
In recent years, bacterial infections are increasing due to emergence of antibiotic resistance.
Oxidative stress and
gall bladder stones also pose a great health threat to millions. This study attempted the assessment of anti-
bacterial, an
ti-oxidant and anti-gall bladder stones activity of an important medicinal plant Achyranthes aspera
methanolic leaf extract. Anti-bacterial activity was performed by disc diffusion method. Significant susceptibility
was observed against gram positive bacteria than gram negative strains. Anti-oxidant activity was studied in terms of
1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging potential with ascorbic acid as standard. Highest
DPPH scavenging activity for A. aspera and ascorbic acid was 59.21% and 92.41%, respectively. IC50 value was
472.93 µg/ml for A. aspera and 1.4965 µg/ml for ascorbic acid. Effects of A. aspera leaf extract on gall bladder
stones were also investigated. 2.0 mg/ml extract concentration showed the highest amount of cholesterol release
(39.69 mg/dl) and the highest amount of dry weight reduction (2.3 mg) from gall stones. Morphological changes like
color change and fragility in gall stones were also observed at concentrations higher than 1.5 mg/ml.
Keyword: Medicinal Plant, Achyranthes aspera, Anti-bacterial, Anti-oxidant, Gall bladder stone, DPPH, IC50
1. Introduction
Bangladesh harbors a large number of
medicinal plants but therapeutic potential of
many of these important medicinal plants
are yet to be revealed. Medicinal plants and
their formulations are used enormously for
treating a range of illness in ethnic medical
practices as well as traditional system of
medicine in India[1]. Since the beginning of
world history, nearly all culture and
civilization depended either fully or partially
on plant derived medicines because these
drugs are effective, low-cost, readily
available and believed to be safer and less
toxic[2]. For these reasons, herbal medicines
and medicinal plants are not only meeting
treatment needs in developing countries but
also getting popular in developed world[3].
Achyranthes aspera is a perennial herb
belonging to the family of Amaranthaceae.
It grows throughout the tropical and warmer
regions of the world[4]. It was reported as an
invasive alien species in northern
Bangladesh[5]. Achyranthes aspera was
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reported to contain many phytochemicals
like alkaloids, flavonoids, tannins,
terpenoids, saponins, glycosides, steroids
etc. Efficacy of Achyranthes aspera was
also established by many scientific
investigations as anti-microbial[6,7],
hypoglycemic[8], cancer chemo-
preventive[9], hepatoprotective[10],
analgesic[11], anti-pyretic, anti-inflammatory
and anti-arthritic[12], hypolipidemic[13],
nephroprotective[14], diuretic[15] and
immunomodulatory[16] etc.
Bacterial infections are great health concern
worldwide. Emergence of antibiotic
resistance and multidrug resistance in recent
years is increasing the incidence of
infections. Infectious diseases are the
world's leading cause of premature
deaths[17]. Therefore, there is a continuous
and urgent need to discover new
antimicrobial compounds with diverse
chemical structures and novel mechanisms
of action. Medicinal plants represent a rich
source of antimicrobial agents. In recent
years, there is a growing interest to evaluate
plants possessing antibacterial activity[18]
and it has also been proved that various
medicinal plants extracts possess
bacteriostatic and bactericidal effects[19].
Natural compounds in plants such as
tannins, terpenoids, alkaloids, and flavonoids
have been found to have antimicrobial
properties established through in vitro
studies.
Antioxidants are type of molecules that
neutralize harmful free radicals, produced
through a chain of reactions[20], that damage
living cells, spoil foods, degrade materials
such as rubber, gasoline, lubricating oil.
Antioxidants terminate these chain reactions
through the removal of free radical
intermediates and inhibition of other
oxidation reactions[21]. The use of
antioxidants in pharmacology is intensively
studied as oxidative stress might be an
important part of many human diseases
particularly stroke and neurodegenerative
incidents[20]. Recently, there has been an
increasing interest in the therapeutic
potentials of medicinal plants as
antioxidants in reducing such free radical
induced tissue injury. Many new plant
species have been investigated in the search
for novel antioxidants[22,23] but there is still a
demand to find more information on the
antioxidant potential of plant species.
Gall bladder is an important organ in human
body exclusively employed in storing biles
secreted from the liver and passes this bile in
response to a fat rich diet. When this bile
contains high level of cholesterol it becomes
hardened, crystalline and doesn’t move from
gall bladder to other parts that is then termed
as gall stones[24]. Gall bladder stones are
mainly cholesterol stones, while pigment
stones and mixed stones composed of bile
pigments and bile salts are also seen[25].
Gallstones incidence is more common in
western society[24]. An estimated twenty
million Americans have gallbladder
problems and approximately 750,000 of
them will have their gallbladder removed
each year[26]. About 80% of all gallstones
are cholesterol stones that vary in color from
light-yellow to dark-green or brown and are
oval 2 to 3 cm in length, contain at least
80% cholesterol by weight (or 70%,
according to the Japanese classification
system) and remaining 20% is bilirubin27.
Many believe that it is possible to treat
gallstones with herbal products and they find
it low cost, safe and painless. Medicinal
plants with anti-oxidant, anti-inflammatory,
anti-bacterial, diuretic and sedative
properties are a good candidate for gall
stone natural treatments.
In this study, Achyranthes aspera
methanolic leaf extract were tested for anti-
bacterial and anti-oxidant activity while
efficacy of this plant extract against gall
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bladder stones were also examined under in
vitro conditions.
2. Materials and Methods:
2.1Collection of Plant and Preparation of
Plant Extract
The fresh and healthy leaves of Achyranthes
aspera were collected from Chittagong
University Campus area. The plant was
taxonomically classified and authenticated
by standard taxonomical method. The fresh
leaves of plant of Achyranthes aspera was
washed with clean water immediately after
collection. The collected leaves were
chopped into small pieces, sun dried for
about 5 days and grinded into coarse powder
with a mechanical grinder and stored in an
airtight container. 166 gm powder was
macerated in 700ml 95% pure methanol
(Sigma Chemicals Co., USA) for 5 days at
room temperature 25±20 C with occasional
stirring. After 5 days, methanol extract was
filtered with Whatman No.1 filter paper. The
extract was concentrated under reduced
pressure below 500C through rotary vacuum
evaporator[28]. The concentrated extract was
collected in a Petri dish and allowed to air
dry for complete evaporation of methanol.
The whole process was repeated three times
and finally, 14.77gm blackish-green colored,
concentrated plant extract was obtained
(yield 8.9% w/w) which was kept in
refrigerator at 40C. The extract thus obtained
is ready for subsequent therapeutic
assessments.
100
X
powder coarse ofamount Total
extract particular ofWeight
ncalculatio Yield
2.2 Anti-Bacterial Activity of A. Aspera
Leaf Methanol Extract
2.2.1 Bacterial strains used in the study
The microorganisms employed in the
current study (Table 1) were procured from
the Laboratory of Molecular Genetics, Dept.
of Genetic Engineering and Biotechnology,
University of Chittagong.
2.2.2 Preparation of discs
The discs of about 4 mm diameter were cut
by punching machine from What man No.1
filter paper. The discs were taken in a Petri
dish and sterilized by autoclave, dried in
oven at 800C.
2.2.3 Preparation of Plant Extracts
Solution
1.0 gm of Achyranthes aspera extract was
accurately weighed and dissolved in 10 ml
of DMSO to give solution of known
concentration (100µg/µl). DMSO was
chosen as solvent because, in addition to that
it can be used in dissolving the crude extract
completely and it has no inhibitory effect on
the cultures.
Table 1: List of Gram (+) ve and Gram (-) ve
bacteria used for the study
Bacterial type
Test organism
Gram (+)ve
Bacillus cereus
Bacillus subtilis
Bacillus megaterium
Staphylococcus sp.
Gram (-)ve
Vibrio cholerae
Shigella sonnei
Shigella dysenteriae
E. coli
Salmonella typhi
Pseudomonas
sp.
2.3 Standard and Media Used
Cefixime antibiotic disc (HiMedia Pvt. Ltd,
Mumbai India) with concentration of
5μg/disc was used as standard in the present
study. Mueller Hinton agar media no. 173
was purchased from Hi media Pvt. Ltd.,
Mumbai, India and the solvents used was
analytical reagent (AR) grade and they were
distilled before use.
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2.4 Disc diffusion Method
The in vitro sensitivity of the bacteria to the
test materials was done by disc diffusion
method well established by Iennette
(1985)[29]. It is essentially a quantitative and
qualitative test indicating the sensitivity or
resistance of the micro-organisms to the test
materials and the extent of sensitivity as
well.
2.5 Determination of Anti-oxidant activity
of A. aspera leaf extract
2.5.1 Working procedures
The antioxidant activity of the plant extracts
and the standard was assessed on the basis
of the scavenging potential of the stable 1,
1-diphenyl-2-picrylhydrazyl (DPPH)-free
radical by modified method[30]. Ascorbic
acid (Sigma Aldrich chemical co. USA) was
used as standard anti-oxidant. Seven
different concentrations (10, 20, 40, 80, 160,
320 and 640 µg/ml respectively) of plant
extract and ascorbic acid were used in this
study. 100 mg of A. aspera leaf extract was
dissolved in 20 ml of pure (95%) methanol
to give final concentration of 5 mg/ml. Then
further dilutions from this stock were also
prepared in methanol for each concentration
of extract tested. Ascorbic acid solution was
prepared in the same way and same
concentration that of the plant extrac. The
diluted working solutions of the test extracts
and ascorbic acid were prepared in
methanol. 0.002% of DPPH was prepared in
methanol. For each concentration, 1 ml of
DPPH solution was mixed with 1 ml of
sample solution. For standard, 1 ml of
DPPH solution was mixed with 1 ml of
ascorbic acid. For, negative control or blank,
one ml of DPPH solution was mixed with 1
ml of methanol to see whether the solvent
used has any anti-oxidant activity or not.
These solution mixtures were kept in dark
for 30 min and shaken vigorously, and
optical density was measured at 517 nm
using Cecil-Elect UV Spectrophotometer.
For each concentration, the optical density
of both the sample and standard was
recorded against the blank and % inhibition
of DPPH free radical was calculated using
the formula given below[31]. All experiments
were performed in triplicate.
Percent (%) inhibition of DPPH = 100 X
A
B-A
Where A = optical density of the blank and B =
optical density of the sample.
Then, log C for each concentration of plant
extract and ascorbic acid was calculated and
plotted in the graph against corresponding %
of DPPH scavenging activity. Then, IC50
(Inhibitory concentration at which 50%
DPPH are neutralized) value for both plant
extract and ascorbic acid was calculated by
regression analysis and using statistical
software “Biostat 2009”.
2.6 Assessment of A. Aspera Leaf Extract
Activity on Gall Bladder Stones
2.6.1 Working protocol
The gall bladder stones were dried at 450C
in an oven and dry weight of the stones was
accurately measured in an air tight 4-digit
electronic balance. Then cholesterol content
in plant extract (blank) was estimated at all
ten concentrations before treating stones.
For this purpose, 0.1 ml from each extract
concentration was added to 9.9 ml of the
FeCl3-CH3COOH solution into a glass
stoppered centrifuge tube. Then it was
mixed well and centrifuged at 4000 rpm for
15 minutes. After centrifugation, it was
stood for 10 minutes for any proteins to
flocculate (if any). Then from the
supernatant 5 ml of clear solution was taken
in a test tube. For the standard, 5 ml of
cholesterol solution was taken in another test
tube. As blank 5 ml of the FeCl3-CH3COOH
solution was taken in a separate test tube.
Then 3 ml of concentrated H2SO4 were
added to all test tubes, mixed by shaking
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carefully. After 30 minutes, the optical
density (O.D) was recorded in UV
spectrophotometer for all the extract
concentrations and standard against the
blank at 560 nm. There were two test tubes
for each sample concentrations and standard.
Afterwards, 5 ml of each extract
concentration was taken in a test tube and
pre-weighed stones were put in the sample.
As blank, 5 ml of distilled water was taken
in a separate test tube. There were 3 test
tubes for each concentration and one stone
in each tube. Then, these tubes with stones
were incubated at 370C for 7 days. After
incubation, the stones were picked up, dried
and again dry weight were taken. Difference
in this dry weight and dry weight before
treatment were calculated to see weight
reduction due to extract effect on stones.
Then, the cholesterol content of the extracts
after treating stones was again measured in
the same procedure as described earlier for
measuring cholesterol level in plant extract
before treating stones. From the difference
in the cholesterol content after and before
treatment were calculated to see the amount
of cholesterol released from the stones due
to effect of the extract.
3. Results and Discussions
3.1 Antibacterial assay of Achyranthes
aspera leaf methanol extract
Antibacterial activity of Achyranthes aspera
leaf methanol extract was studied against
four Gram positive and six Gram negative
bacteria by disc diffusion method and
compared with the standard antibiotic disc
of Cefixime (5μg/disc). DMSO 2mg/disc
was used as negative control. Antibacterial
activity of Achyranthes aspera leaf
methanol extract was tested at three different
concentrations such as 1 mg/disc, 2 mg/disc
and 3 mg/disc. All three concentrations
produced zone of inhibition and thus showed
different degree of antibacterial activity
against all ten strains of the bacteria. It was
observed that gram positive bacteria showed
slightly greater susceptibility than gram
negative bacteria to the plant extract. A dose
dependent antibacterial activity was also
found. With the increase in extract
concentration, the zone of inhibition was
also increased. However, the highest zone
of inhibition was observed in 3 mg/disc
extract for all the strains. For 3 mg/disc,
zone of inhibition was the highest (13.5 mm)
in Bacillus cereus and Bacillus megaterium
and the lowest (7.5 mm) in Pseudomonas sp.
For 2 mg/disc, zone of inhibition was
highest (10.5 mm) in Bacillus cereus and the
lowest (6.5 mm) in Salmonella typhae. For
1 mg/disc, zone of inhibition was the highest
(8.5 mm) in Bacillus megaterium and the
lowest (5.0 mm) in three strains namely,
Staphylococcus aureus, E. coli and
Salmonella typhae. Disc with DMSO
showed no zone of inhibition at all. Standard
antibiotic disc of cefixime also exhibited no
susceptibility to the strains tested. (Table 2).
An inhibition zone of 10mm or greater was
considered to indicate good antibacterial
activities[32]. Even low concentration of
plant extract (1 mg/ml) also inhibited
bacterial growth in Bacillus megaterium and
Bacillus cereus (8.5 and 7.5 mm
respectively). This result is proved to be
better in compared to previous study
of[32,33,34]. Antibacterial study in other
species of this genera also showed poor
response and required high dose[35,36,37].
Table 2: In vitro antibacterial activity of A. aspera leaf methanol extract
Type of
organism Bacterial strain
Diameter of zone of inhibition (mm)
Different conc. of plant extract Cefixime
(standard)
DMSO
(Negative
control)
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1mg/disc 2mg/disc 3mg/disc 5µg/disc 2mg/disc
Gram (+)ve
Bacillus cereus
7.5
10.5
13.5
0
0
Bacillus subtilis
6.5
8.5
9.0
0
0
Bacillus
megaterium 8.5 9.0 13.5 0 0
Staphylococcus
aureus 5.0 7.0 8.5 0 0
Gram
(-) ve
Vibrio cholerae
6.5
8.5
10.0
0
0
Shigella sonnei
6.0
8.0
9.5
0
0
Shigella dysenteriae
6.0
8.0
9.5
0
0
E. coli
5.0
8.0
8.5
0
0
Salmonella typhae
5.0
6.5
8.5
0
0
Pseudomonas sp.
6.0
7.0
7.5
0
0
3.2 Determination of in vitro Anti-Oxidant
Activity of A. aspera leaf Methanol
Extract
The scavanging capability of DPPH was
determined by the decrease in its absorbance
at 517 nm and also by the degree of colour
change from purple to yellow. Both ascorbic
acid and A. aspera methanol extract showed
dose dependent activity. Among the seven
different concentrations tested in this study
(10, 20, 40, 80, 160, 320 and 640 μg/ml)
ascorbic acid showed 60.40, 69.80, 75.44,
80.18, 83.56, 87.30 and 92.41% DPPH free
radical scavenging activity respectively,
where 640μg/ml concentration produced
highest scavenging activity (92.41%) . For
all the seven concentrations, A. aspera
methanolic leaf extract showed 17.86, 25.41,
32.59, 34.56, 35.79, 42.0 and 59.21%
scavenging activity respectively, where the
highest DPPH scavenging activity was
59.21% for A. aspera methanol extract at the
concentration of 640μg/ml. It was observed
that % of DPPH free radical scavenging
activity linearly increased with the increase
in concentrations for both ascorbic acid and
A. aspera leaf extract. The anti-oxidant
activity in terms of % of DPPH free radical
scavenging activity or % of inhibition was
plotted against log concentrations for both
ascorbic acid and A. aspera leaf extract and
from the graph IC50 (Inhibition
concentration 50) value was calculated by
linear regression analysis. IC50 value of
ascorbic acid and A. aspera leaf methanol
extract was found to be 1.4965 and 472.93
μg/ml, respectively (Fig.1 and 2). The graph
in Fig. 3 shows comparison of anti-oxidant
activity between ascorbic acid and A. aspera
leaf extract.
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Fig. 1: Regression line for determining the IC50 value of Ascorbic acid
Fig. 2: Regression line for determining the IC50 value of Achyranthes aspera leaf methanol extract.
y = 16.45x + 47.12
R² = 0.970
Log IC 50 = 0.1751…
0
20
40
60
80
100
0 1 2 3
% of DPPH Scavanging Activity
Log Concentration
% of DPPH Scavanging Activity of Ascorbic Acid
DPPH Scavanging Activity
-Liner (Regression Line)
y = 18.99x - 0.794
R² = 0.896
IC 50 = 472.93 µg/ml)
Log IC 50 = 2.6748
0
10
20
30
40
50
60
70
0123
% of DPPH scavanging activity
Log Concentration
% of DPPH scavanging activity of A. aspera leaf extract
DPPH scavanging activity
-Liner (Regression Line)
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Fig.3: Comparison of anti-oxidant activity of A. aspera and Ascorbic acid
A good linearity was observed for regression
line where R2 value was 0.970 and 0.896 for
ascorbic acid and A. aspera extract
respectively (Fig. 1 and 2). The reduction
capability of DPPH was determined by the
decrease in its absorbance at 517 nm and
also by the colour change from purple to
yellow, which is induced by anti-
oxidants[38]. The degree of discoloration
indicates the scavenging potential of the
antioxidant compound in the extracts. This
result is in conformity with that previously
obtained by other studies[33,39]. However, the
chemical constituents present in the extract,
which are responsible for anti-oxidant
activity, need to be investigated. The
phytochemical tests indicated the presence
of alkaloids, glycosides, tannins, and
flavonoids in the crude methanolic extract.
Several of such compounds are known to
possess potent antioxidant activity[40]. The
majority of the antioxidant activity is due to
the flavones, isoflavones, flavonoids,
anthocyanin, coumarin lignans, catechins
and isocatechins[41]. In in vitro condition,
such information may be of potential value
in the design of further studies to unravel
novel strategies for disorders associated with
free radicals-induced tissue damage[39].
3.3 Assessment of Gall Bladder Stones
Treatment
Gall bladder stones collected from hospitals
(operation theatre) were treated with ten
different concentrations of A. aspera extract
(0.0625, 0.125, 0.25, 0.5, 0.75, 1.0, 1.25,
1.50, 1.75 and 2.0 mg/ml). Distilled water
without extract was used as negative control.
Stones were categorized as cholesterol
stones (yellowish or brown color) and
pigment stones (dark, blackish colored).
First, the cholesterol content of the extract
concentrations before treating stones were
estimated. Very low level of cholesterol was
found in the extract. It was observed that,
with the increase in concentrations the
amount of cholesterol was also found to be
increased. The highest amount of cholesterol
was obtained in 2.0 mg/ml concentration
and lowest amount was obtained in 0.0625
mg/ml.
Then, amount of cholesterol in different
concentrations of extract were again
measured after treating gall bladder stones in
extract for 7 days period at 370C. It was
observed that, the amount of cholesterol was
higher in all concentrations after treatment
than that obtained before treatment which
0
10
20
30
40
50
60
70
80
90
100
0 100 200 300 400 500 600 700
% 0f DPPH Scavanging
Activity
Concentrations (µg/ml)
Comparative anti-oxidant activity of A. aspera and Ascorbic Acid
A. asperaLeaf extract
Ascorbic Acid
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indicates that some cholesterol must have
been released in the extract from stones.
From the difference between amount of
cholesterol in extract before treatment and
after treatment, the amount of cholesterol
released was calculated for each
concentration and presented in Fig. 4. With
the increase in extract concentrations, the
amount of cholesterol release also increased.
The highest amount of cholesterol was
released (39.69 mg/dl) from stone treated
with 2.0 mg/ml extract concentration and
lowest amount of cholesterol released
(23.55mg/dl) was observed in 0.0625 mg/ml
extract concentration whereas in the control
or distilled water only 7.80 mg/dl
cholesterol released.
The effect of the A. aspera extract on the dry
weight of the stones before and after
treatment was also examined. It was
observed that the plant extract has negligible
effect on the dry weight. Very minute
amount of weight reduced after treatment.
However, the amount of weight reduced was
increased with increasing concentrations of
extract. The maximum weight reduction was
2.3 mg observed in 2.0 mg/ml extract
concentration while the control or distilled
water reduced only 0.3 mg. This result of
weight reduction is presented in column
chart (Fig. 5). Morphological changes were
also observed in stones after treating with
plant extract. The color of cholesterol stones
was changed from yellowish brown to
white. Concentrations higher than 1mg/ml
induced color change. No color change
observed in dark or pigment stones. The
stones were much harder before treatment
and fragility was observed after treating with
extract. Only the stones treated with extract
concentrations of 1.50, 1.75 and 2.0 mg/ml
showed fragility in a lesser extent.
Fig.4: Column chart showing amount of cholesterol released from gall bladder stones after treating with A. aspera
extract at different concentrations
0
5
10
15
20
25
30
35
40
45
Amount of cholesterol released
(mg/dl)
A. aspera leaf extract concentrations (mg/ml)
Amount of cholesterol
released from stones
(mg/dl)
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Fig.5: Column chart showing the weight reduction of gall bladder stones after treating with A. aspera extract at
different concentrations.
Achyranthes aspera is the plant, which is
commonly used in Ayurveda for the
treatment of stomachache, digestive
problems, piles, boils and skin erruptions[42].
A. aspera was reported to inhibit
mineralization of urinary stones (calculi)
like calcium oxalate, calcium carbonate and
calcium phosphate[43]. Methanolic extracts
were found to prevent lead induced
nephrotoxicity in albino rats[14]. Efficacy of
the roots of the plant was tested on
nucleation of calcium oxalate crystal and
growth in vitro and on oxalate induced
injury in NRK-52E (rat renal tubular
epithelial) cells[44]. As an approach to anti-
lithiasis, inhibitory effect of hydroalcoholic
extract of the plant on crystallization of
calcium oxalate in synthetic urine was
investigated[45]. This plant was reported to
contain saponins. Saponins can emulsify
cholesterol and fat present in the stones.
Diuretic property of the plant also supposed
to contribute in gall bladder stone treatment
as it can increase the volume of bile thus
preventing stone formation[15]. Antibacterial
activity of the plant is also important in
applying it for gall stone treatment to
prevent infection. Anti-oxidant activity of
the plant is also helpful in preventing
oxidative stress to gall bladder[46]. Anti-
inflammatory activity of A. aspera was
reported earlier[47,12] supposed to reduce pain
caused by cholicystitis. However, in vitro
treatment of gall stones with medicinal plant
extract was not previously attempted. No,
established report was available in this
regard. This protocol was a very preliminary
approach which needs more in vitro and in
vivo studies to reach a final conclusion.
4. Conclusion
This study observed an appreciable degree
of anti-bacterial and anti-oxidant activity of
Achyranthes aspera methanolic leaf extract
as well as it’s efficacy against gall bladder
stones was also promising. Its effect on gall
stones provided totally new findings and
opened new window to think about the
potentials of this plant. However, the
identification, isolation and purification of
active phytochemical constituents
responsible for these therapeutic properties
may lead to new drug development from this
plant. The therapeutic activities tested in this
study on A. aspera leaf extract had shed
more light on pharmacological importance
Journal of Medicinal Plants Studies www.plantsjournal.com
Vol. 1 Issue. 3 2013 www.plantsjournal.com Page | 115
of this plant and thus further in vitro and in
vivo studies can help large number of people
to utilize the natural medication from this
plant.
5. Acknowledgements
The authors acknowledge to Dr. Dwaipayan
Sikdar, Associate Professor, Dept. of
Biochemistry and Molecular Biology,
University of Chittagong for his help in
extract preparation, to Dr. Md. Nur Hossain
Bhuiyan (Shaheen), Assistant Professor,
Dept. of Surgery, Chittagong Medical
College, Chittagong, Bangladesh for
supplying gall bladder stones and finally to
the Ministry of Science and Technology,
Government of the People’s Republic of
Bangladesh for financial support.
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- CitationsCitations0
- ReferencesReferences41
- [Show abstract] [Hide abstract] ABSTRACT: Solvent leaf extracts of Achyranthes aspera Linn. was tested for its antibacterial and antifungal activities against the organisms, E. coli, Enterobacter sp. P. aeruginosa, P. vulgaris, S. aureus, Klebsiella sp. Salmonella sp. Shigella sp. Trichophyton mentagrophytes, T. rubrum, T. tonsurans, Aspergillus sp. E. floccosum. C. neoformans and Candida albicans. In the present study, maximum inhibitory activity was observed in diethyl ether extract against all the bacterial and fungal species.
- [Show abstract] [Hide abstract] ABSTRACT: The methanolic crude extracts of 12 traditionally used Indian medicinal plants were screened for their antioxidant and free radical scavenging properties using α-tocopherol and butylated hydroxy toluene (BHT) as standard antioxidants. Antioxidant activity was measured by ferric thiocyanate (FTC) assay and compared with the thiobarbituric acid (TBA) method. Free radical scavenging activity was evaluated using diphenyl picryl hydrazyl (DPPH) radicals. The overall antioxidant activity of Lawsonia inermis was the strongest, followed in descending order by Ocimum sanctum, Cichorium intybus, Piper cubeba, Punica granatum, Allium sativum, Delonix regia, Terminalia chebula, Terminalia bellerica, Mangifera indica, Camellia sinensis, and Trigonella foenum-graecum. Seven plants, namely Terminalia chebula, Mangifera indica, Terminalia bellerica, Punica granatum, Ocimum sanctum, Cichorium intybus, and Camellia sinensis, showed strong free radical scavenging activity with the DPPH method. Phytochemical analysis of plant extracts indicated the presence of major phytocompounds, including phenolics, alkaloids, glycosides, flavonoids, and tannins. The phenolic concentrations in the above plants ranged from 28.66 to 169.67 mg/g of dry plant extract. A fair correlation between antioxidant/free radical scavenging activity and phenolic content was observed among 9 plants; however, in 3 plants (Piper cubeba, Lawsonia inermis and Trigonella foenum-graecum), no such relationship was observed. The tested plant extracts showed promising antioxidant and free radical scavenging activity, thus justifying their traditional use.
- [Show abstract] [Hide abstract] ABSTRACT: The inhibition of mineralization of urinary stone forming minerals by medicinal plants i.e . Achyranthes aspera Linn, Passiflora leschenaultii DC, Solena amplexicaulis (Lam.) Gandhi, Scoparia dulcis Linn and Aerva lanata (Linn.) been investigated. The inhibition efficiency was studied. Increased intake of fruits juice and seed extract of our plants would be helpful in urinary stone prophylaxis .
- [Show abstract] [Hide abstract] ABSTRACT: The present study was designed to evaluate the nephroprotective role of methanolic extract of Achyranthes aspera (A.aspera) an important herb in the Indian system of medicine against lead acetate-induced nephrotoxicity in rats. Toxicity was induced in male albino rats (Wistar strain) by administering lead acetate (0.2%) in drinking water for 6 weeks, followed by extract of A. aspera (200 mg/kg body weight). Changes in kidney weights encountered upon lead administration improved after extract with A. aspera. Lead damage to the urine was evident from increase in the activity of γ-glutamyltranspeptidase (γ-GT), Cathespin D, alkaline phosphatase (ALP), acid phosphatase (ACP), β-glucuronidase lactate dehydrogenase (LDH) and N-acetyl-β-D-glucosaminidase (NAG) in urine along with some urinary constituents (urea, uric acid, creatinine, protein and phosphorous). The effects of lead were also studied in kidney (γ-GT, β-glucuronidase, NAG, Cathespin D and LDH) and showed a decline upon extract administration. Increased activities of urinary enzymes were accompanied by increase in the urinary constituents. Treatment with methanolic extract of A. aspera after lead induction completely ameliorated the lead-induced renal damage.
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