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Authors' contributions This work was carried out in collaboration between all authors. Authors MI and QA designed the study, performed the statistical analysis. Authors MRK and MAR wrote the protocol and wrote the first draft of the manuscript. Authors AK and MI managed the analyses of the study. Author RBR managed the literature searches. Author MAR revised manuscript. All authors read and approved the final manuscript. ABSTRACT Aims: The crude methanol extract of whole plant of Blumea lacera (Burn.f.) DC. has been investigated for anti-diarrheal, antimicrobial, anxiolytic, anti-atherothrombosis, membrane stabilizing and alpha-amylase inhibitory activities. Place and Duration of Study: The study was carried out in 2013 in the Department of Pharmacy, Southern University Bangladesh, Chittagong, Bangladesh. Methodology: Test for anti-diarrheal activity was carried out by castor oil-induced diarrhea in mice. The preliminary antimicrobial activity was determined by the agar disc diffusion method. The anxiolytic activity was examined in mice by using the hole board test and open field test (OFT). The anti-atherothrombosis activity was evaluated using standard streptokinase. The membrane stabilizing activity was assessed by using hypotonic solution induced hemolysis of human erythrocyte. The plant extract was also assessed for anti-diabetic ability using In vitro α-amylase inhibitory potential. The α-amylase inhibitory activity of B. lacera was measured using the starch-iodine method. 1678 Results: The crude extract of B. lacera showed anti-diarrheal activity in dose-dependent manner. In antimicrobial assay, this extract showed better activity against the tested fungi compared to the bacteria used in the screening. Significant anxiolytic activity was found for this plant extract. In the In vitro anti-atherothrombosis test, the extract exhibited 46.17% clot lysis as compared to the standard, streptokinase (81.53%). In membrane stabilizing activity test, the plant extract at 1.0mg/ml inhibited the heat-induced hemolysis of RBCs by 52.27% whereas the standard acetyl salicylic acid (ASA) demonstrated 81.72% inhibition of hemolysis. Our results revealed that the extract had dose dependent prevention of digestion of carbohydrates by inhibiting α-amylase. The ability of B. lacera to inhibit thermal-and hypotonic-enzyme activity was found to be statistically significant (p=0.05). Conclusion: These results demonstrated that B. lacera may be used in pharmaceutical applications because of its effective pharmacological properties.
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*Corresponding author: Email: rashidma@du.ac.bd;
British Journal of Pharmaceutical Research
4(13): 1677-1687, 2014
SCIENCEDOMAIN international
www.sciencedomain.org
Pharmacological Activities of Blumea lacera
(Burm. f) DC: A Medicinal Plant of Bangladesh
Abul Khair
1
, Mohammed Ibrahim
1
, Qamrul Ahsan
1
, Zilly Homa
1
,
Md. Ruhul Kuddus
2
, Ridwan Bin Rashid
3
and Mohammad Abdur Rashid
2*
1
Department of Pharmacy, Southern University Bangladesh, Chittagong, Bangladesh.
2
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Dhaka,
Dhaka-1000, Bangladesh.
3
Department of Microbiology, University of Dhaka, Dhaka-1000, Bangladesh.
Authors’ contributions
This work was carried out in collaboration between all authors. Authors MI and QA
designed the study, performed the statistical analysis. Authors MRK and MAR wrote the
protocol and wrote the first draft of the manuscript. Authors AK and MI managed the
analyses of the study. Author RBR managed the literature searches. Author MAR revised
manuscript. All authors read and approved the final manuscript.
Received 11
th
March 2014
Accepted 19
th
April 2014
Published 7
th
July 2014
ABSTRACT
Aims:
The crude methanol extract of whole plant of Blumea lacera (Burn.f.) DC. has been
investigated for anti-diarrheal, antimicrobial, anxiolytic, anti-atherothrombosis, membrane
stabilizing and alpha-amylase inhibitory activities.
Place and Duration of Study: The study was carried out in 2013 in the Department of
Pharmacy, Southern University Bangladesh, Chittagong, Bangladesh.
Methodology: Test for anti-diarrheal activity was carried out by castor oil-induced
diarrhea in mice. The preliminary antimicrobial activity was determined by the agar disc
diffusion method. The anxiolytic activity was examined in mice by using the hole board
test and open field test (OFT). The anti-atherothrombosis activity was evaluated using
standard streptokinase. The membrane stabilizing activity was assessed by using
hypotonic solution induced hemolysis of human erythrocyte. The plant extract was also
assessed for anti-diabetic ability using In vitro α-amylase inhibitory potential. The α-
amylase inhibitory activity of B. lacera was measured using the starch-iodine method.
Original Research Article
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1678
Results:
The crude extract of B. lacera showed anti-diarrheal activity in dose-dependent
manner. In antimicrobial assay, this extract showed better activity against the tested fungi
compared to the bacteria used in the screening. Significant anxiolytic activity was found
for this plant extract. In the In vitro anti-atherothrombosis test, the extract exhibited
46.17% clot lysis as compared to the standard, streptokinase (81.53%). In membrane
stabilizing activity test, the plant extract at 1.0mg/ml inhibited the heat-induced hemolysis
of RBCs by 52.27% whereas the standard acetyl salicylic acid (ASA) demonstrated
81.72% inhibition of hemolysis. Our results revealed that the extract had dose dependent
prevention of digestion of carbohydrates by inhibiting α-amylase. The ability of B. lacera to
inhibit thermal-and hypotonic-enzyme activity was found to be statistically significant
(p=0.05).
Conclusion: These results demonstrated that B. lacera may be used in pharmaceutical
applications because of its effective pharmacological properties.
Keywords: Blumea lacera; anti-diarrheal; antimicrobial; anxiolytic; anti-atherothrombosis;
membrane stabilizing; alpha-amylase.
1. INTRODUCTION
The use of natural products or natural product-based medicine is increasing all over the
world especially in the developing countries such as Bangladesh, India, China, and the
Middle East. About 25% of the prescribed drugs in the world are of plant origin [1].
Approximately 80% people rely on traditional plant-based drugs for their primary health care
needs in the developing countries [2]. Since ancient times, different parts of medicinal plants
have been used for ailments caused by microorganisms. There is wide range of medicinal
plant parts possessing a variety of pharmacological activities; such are used as powerful raw
drug. Recent widespread interest in plant-derived drugs reflect its recognition of the validity
of many traditional claims regarding the values of natural products in health care [3]. For
quality control of traditional medicines, phytochemical screenings are mainly applied. Now a
days, secondary plant metabolites with previously unknown pharmacological activities have
been extensively investigated as source of medicinal agents. Thus, it is anticipated that
phytochemicals with enough antibacterial efficacy will be used for the treatment of bacterial
infections [4]. According to WHO, medicinal plants are the best sources to obtain a variety of
new herbal drugs. Therefore, in order to determine the potential use of herbal medicine, it is
important to emphasize the study of medicinal plants that found in folklore [5].
Blumea lacera (Burn.f.) DC. (Bengali name: Kukursunga; Family-Asteraceae) is an erect
herb which grows as a weed in uncultivated lands all over Bangladesh. The alcoholic extract
of the herb exhibited marked anti-inflammatory activity against carrageenin- and bradykinin-
induced inflammation in rats. Essential oil from leaves have analgesic, hypothermic and
tranquillizing activities [6]. The plant also exhibited anti-leukemic, antiviral [7] and cytotoxic
[8] activities against breast cancer cells.
As part of our ongoing efforts to study medicinal plants of Bangladesh [9-11], we evaluated
the anti-diarrheal, antimicrobial, anxiolytic, anti-atherothrombosis, membrane stabilizing and
alpha-amylase inhibitory activities of B. lacera as well as to find out the logical evidence for
its folk uses of this plant as described in Ayurveda. Although in an earlier experiment, the
ethanol extract of root of B. lacrera has been reported to exhibit the anti-diarrheal activity
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1679
[12], but the present study was designed to evaluate the pharmacological activities of crude
methanol extract of whole plant of this plant.
2. MATERIALS AND METHODS
2.1 Collection of Plant Materials
Leaves of B. lacera were collected from Chittagong Pahartoli, Bangladesh in June 2012 and
were identified at the Forest Research Institute, Chittagong, Bangladesh where a voucher
specimen has been maintained for future reference.
2.1.1 Drying and grinding
After collection, the leaves were washed with running tap water. These clean leaves were
dried at a temperature not exceeding 50ºC. The dry materials were ground to a coarse
powder with the help of a grinder and kept in an airtight container. The container was then
stored in a cool and dark place until extraction was commenced.
2.1.2 Hot extraction by soxhlet extractor
Exactly 140gm of powdered leaf was extracted with 750ml of methanol (99.98%) with a
Soxhlet apparatus (Quickfit, England). The extract was concentrated with a rotary evaporator
(Heidolph, Germany) under reduced temperature and pressure to provide a gummy residue
(yield 18.70%).
2.1.3 Chemicals
All chemicals and solvents used in this study were of analytical grade and purchased from
Merck, Germany. Standard drugs such as loperamide, ciprofloxacin, fluconazole, diazepam,
acetyl salicylic acid and acarbose were obtained from Square Pharmaceuticals Ltd as gift
samples.
2.1.4 Experimental animals
For the experiment Swiss albino mice of either sex, 6-7 weeks of age, weighing between 25-
30g, were collected from the Animal Resources Branch of the International Centre for
Diarrheal Disease and Research, Bangladesh (ICDDR,B). The mice were maintained under
standard environmental conditions of temperature: (27.0±1.0ºC), relative humidity: 55-65%
and 12h light/12hr dark cycle and had free access to ICDDR,B formulated diet and water ad
libitum. Appropriate measures were taken to minimize the pain or discomfort of animals and
the mice were acclimatized to laboratory condition for one week prior to experiments.
All protocols for animal experiment were approved by the institutional animal ethical
committee [13].
2.2 Test for Anti-diarrheal Activity
Anti-diarrheal activity of crude methanol extract of B. lacera was assessed by castor oil-
induced diarrhea in mice [14]. The animals were divided into control, positive control and two
test groups containing seven mice in each. Control group received 1% Tween-80 (10ml/kg,
p.o). The positive control group received loperamide (25mg/kg, p.o.) while the test groups
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1680
received the methanol extract (100 and 200mg/kg b.w.) orally. Acute diarrhea was produced
by oral administration of 0.4ml of castor oil in each mouse. The latency period and diarrheic
secretion were counted for 4 hours.
2.3 Test for Antimicrobial Activity
The preliminary antimicrobial activity of the extractive was determined at 500µg/disc by the
agar disc diffusion method [15] against a number of Gram-positive and Gram-negative
bacteria and fungi. The bacterial and fungal strains used in this experiment were collected
from the Microbiology Lab., Chittagong University, Chittagong, Bangladesh. Standard
ciprofloxacin (30µg) and fluconazole (50µg) disc were used as reference.
2.4 Test for Anxiolytic Activity
2.4.1 Treatment schedule
The anxiolytic activity of B. lacera was examined by using the hole board test and open field
test (OFT). The animals were divided in to four groups, with each group consisting of seven
mice. First group received normal saline, second group received diazepam (1mg/kg b.w.,
p.o.), while the third and fourth groups received plant extract at 200 and 400mg/kg b.w.
respectively.
2.4.2 Hole board test
The hole board is a white painted wooden board (30cm×20cm×14cm) with 16 holes (each of
diameter 3cm) evenly distributed on the base of box. The test groups orally received crude
extract of B. lacera at the dose of 200 and 400mg/kg b.w. respectively. The control group
received saline and positive control received diazepam (2mg/kg-i.p.) respectively. The
number of passages of a mouse through the hole from one chamber to the other
was counted for a period of 5min at 30min after oral administration of both doses of the test
drug [16].
2.4.3 Open field test
The open field test is used to observe general motor activity, exploratory behavior and
measures of anxiety. The open field area was made of plain wood and consisted of a square
area (45cm×45cm×20cm). The floor had a square sheet of wood (45cm×45cm) with the
surface divided into sixteen small squares. Mice were divided into four groups of 7 mice and
treated similarly as described in hole cross test. About 30min after treatment, mice of both
the control and treated groups were placed individually at the center of the open field and
behavioral activities were recorded for 5min. Subsequently, hand operated counters and
stopwatches were used to score the following behavioral parameters for a period of 5min: (1)
the number of entries and time spent in the centre, (2) periphery and corners of the field, (3)
the number of crossings (number of square floor units entered) as a measure of distance
traveled, (4) rearing (number of times the animal stood on hind legs) and (5) assisted rearing
(forepaws touching the walls of the apparatus) [16].
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1681
2.5 Test for Anti-atherothrombosis Activity
The anti-atherothrombosis activity of the crude extract was evaluated using streptokinase as
standard [17]. For this study, 4ml venous blood was drawn from healthy volunteers and
distributed in three (for extract, reference standard and for negative control) pre-weighed
sterile micro-centrifuge tubes (0.5ml/tube). The tubes were incubated at 37ºC for 45min.
After clot formation, serum was completely removed without disturbing the clot and each
tube was weighed again to determine the weight of clot (clot weight=weight of clot containing
tube–weight of tube alone). Then, 100µl of methanol extract at a dose of 5µg/µl, 100µl of
streptokinase and 100µl of methanol were separately added to the pre-marked tubes
containing the clot. The tubes were then incubated at 37ºC for 90min and observed for clot
lysis. Afterwards, the fluid released was removed and tubes were again weighed to observe
the difference in weight after clot disruption. Difference obtained in weight taken before and
after clot lysis was expressed as percentage of clot lysis. The experiment was repeated for
three times in different days with fresh blood samples collected from 10 healthy volunteers
(male and female) having no history of contraceptives and anticoagulants.
2.6 Test for Membrane Stabilizing Activity
For this experiment, three clean centrifuge tubes were taken for positive control (acetyl
salicylic acid), three for negative control (99.8% methanol) and six for crude methanol
extract. One milliliter of 10% RBC suspension was added to each tube. Then 1.0ml methanol
and 1.0ml acetyl salicylic acid were added to the negative control and positive control tubes,
respectively. On the other hand, for the test group, 1.0ml of methanol extract (1000mg/kg)
was added. The pH (7.4±0.2) of the reaction mixtures was adjusted by phosphate buffer.
The tubes were incubated in water bath and after cooling, these were centrifuged at
2500rpm for 5min. After filtration the absorbance of the supernatants were measured at
556nm. The total inhibition of hemolysis was then calculated by determining the % inhibition
of hemolysis [18].
2.7 Test For Alpha-amylase Inhibitory Activity
The α-amylase inhibitory activity of B. lacera was measured using the starch-iodine method
[19]. Twenty microliter of α-amylase solution (0.030mg/ml) was mixed with 1.3ml of Tris-HCl
buffer (0.01M containing 0.006M NaCl, pH 6.8) and the crude extract (80, 160, 320µl). After
incubation at 37ºC for 20min, 100µl of the starch solution (0.1%) was added, and the mixture
re-incubated for 20min, after which 2ml of 0.01% acidic iodine solution was added. The
absorbance of the sample was measured at 565 nm and percentage inhibition was
calculated as % Inhibition of enzyme activity =(A-C)×100/ (B-C) where, A=absorbance of the
sample, B=absorbance of blank (no extract), and C=absorbance of control (no starch).
2.8 Statistical Analysis
Results are expressed as the mean±SEM (SEM=Standard Error of Mean). Statistical
analysis for animal experiment was carried out using one-way ANOVA followed by Dunnett’s
multiple comparisons. The results obtained were compared with the vehicle control group;
p=0.05 was considered as statistical significant.
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1682
3. RESULTS
3.1 Anti-diarrheal activity
In the castor oil-induced diarrheal experiment in mice, the crude methanol extract of B.
lacera at 100 and 200mg/kg b.w. significantly reduced the total number of episodes of
defecation as well as delayed the onset of diarrhea in a dose dependent manner Table 1.
These results were shown to be statistically significant (p=0.05).
3.2 Antimicrobial Activity
In the anti-bacterial screening by disc diffusion method, the zones of inhibition were found
within the range of 9.0 to 14.0mm. The highest zone of inhibition (14.0mm) was observed
against Shigella dysenteriae Table 2. But the plant extract also showed mild antibacterial
activity against Salmonella Typhi, Bacillus cereus, Staphylococcus aureus. During the anti-
fungal test, the zones of inhibition were found within the range of 11.0 to 18.17mm with the
highest zone (18.17mm) observed for Blastomyces dermatitidis Table 2.
3.3 Anxiolytic Activity
3.3.1 Hole board test
The number of line crossings was found to increase significantly in case of diazepam treated
animals as compared to control animals. The plant extracts at the 200 and 400mg/kg b.w.
(p.o) dose showed significant increase in the number of line crossing as compared to control
animals as shown in Table 3.
3.3.2 Open field test
Significant anxiolytic activity was observed for diazepam as well as plant extracts when
compared to control. In the open field test, administration of plant extract in mice showed
significant increase in the number of squares crossed during 5min intervals of test as
compared with control as show in Table 3.
3.4 Anti-atherothrombosis Activity
In the anti-atherothrombosis activity test, streptokinase, a positive control (30,000IU) showed
81.53% clot lysis. On the other hand, clots when treated with 100µl methanol (negative
control) showed only negligible lysis (2.87%). In the same time, by treating clots with 500µl
(µg/ml) of the extract, 46.17% clot lysis was obtained. Statistical representation of the
effective clot lysis percentage has been shown in Table 4.
3.5 Membrane Stabilizing Activity
The extract at 500, 1000µg/ml inhibited the heat induced hemolysis of RBCs by 35.90,
52.27%, respectively whereas standard acetyl salicylic acid showed 71.36% inhibition of
hemolysis Table 4. The stabilization of cell membrane for crude methanol extract was found
to be moderate. Although the precise mechanism of this membrane stabilization is yet to be
elucidated, it is thought that the plant might inhibit the release of lysosomal content of
neutrophils at the site of inflammation.
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1683
Table 1. Effect of crude methanol extract of B. lacera on castor oil-induced diarrhea in
mice
Mean latent
period (min)
%
defecation
% Inhibition of
defecation
TNF (240
min)
Control 27.67 100 0 68±1.78
Loperamide (3mg/kg) 73.67 19.12 80.88 13±0.41
BLME (200mg/kg) 58.26 47.06 52.94 46.98±0.81
BLME (100mg/kg) 44.0 69.12 30.88 31.98±0.41
TNF=Total number of feces; Values are mean±SEM (n=7); **p=0.05 by Dunnett’s T test for values
between the sample and vehicle treated group; SEM=Standard Error of Mean; BLME=B. lacera
methanol extract
Table 2. Antimicrobial activity of crude methanol extract of B. lacera at 500µg/disc
Microorganisms
Zone of inhibition (MZI±SD) mm
BLME (500
µg/disc)
Standard
(30
µg/disc)
Gram p
ositive bacteri
a
c
iprofloxacin
Bacillus cereus 11.66±0.68
15.33±0.41
B. megaterium 10.66±0.40
20.67±0.41
B. subtilis 11.33±0.82
15.00±0.71
Staphylococcus aureus 11.67±0.41 18.00±1.41
Gram negative bacteria
Escherichia coli 9.0±0.70 16.67±0.41
Pseudomonas aeruginosa 9.66±0.40 17.00±1.22
Salmonella Typhi 13.66±1.08 15.67±0.41
Shigella dysenteriae 14.0±0.71
15.33±0.82
Fungi
f
luconazole
Aspergillus niger 12.62±0.41
16.5±0.35
Blastomyces dermatitidis 18.17±1.74
19.67±0.54
Cryptococcus neoformans 15.16±1.816
18.5±1.06
Protozoa
fluconazole
Plasmodium ovale 11.00±0.70
15.33±0.74
MZI: Mean zone of inhibition (mm); zone of inhibition under 7mm were considered as less active and
were discarded. Values are expressed as mean±SD (n=3). Means with different letters are significantly
different (p=0.05). BLME=B. lacera methanol extract
Table 3. Anxiolytic effect of crude methanol extract of B. lacera in mice by hole cross
test and open field test
Treatment
Dose
(mg/kg)
Hole cross test
Open field test
Number of
hole crossing
% Inhibition
of hole cross
Number of
square
crossed
% Inhibition
of square
cross
Saline 1ml 25.67±1.08 0.0 251.67±2.94 0.0
Diazepam 1 3.33±0.71 87.01 39.67±2.54 84.24
BLME 200 8.67±0.41 66.23 118.33±1.42 32.67
BLME 400 6.0±0.41 76.62 67.31±1.78 52.99
All values are mean±SEM (n=7). SEM=Standard Error of Mean; BLME=B. lacera methanol extract
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1684
3.6 Alpha-amylase Inhibitory Activity
The extract of B. lacera displayed concentration dependent inhibitory effect on the starch
breakdown In vitro as shown in Table 5. In the present study, the crude extract showed
29.82%, 46.49%, 63.15% inhibition of α-amylase enzyme activity at 80, 180 and 320µg/ml,
respectively whereas the standard acarbose (50µg/ml) produced 79.80% inhibition. The
ability of B. lacera to inhibit and hypotonic and thermal enzyme activity was found to be
statistically significant (p=0.05).
Table 4. Anti-atherothrombosis and membrane stabilization activity of crude methanol
extract of B. lacera
Anti
-
atherothrombosis activity
Membrane stabilizing activity
% Clot lysis
Total inhibition of
hemolysis
Control 2.87±0.94 00.00±0.018
Streptokinase
(positive control) 81.53±3.7 ND
Positive control
(ASA, 0.1mg/ml) ND 71.36±0.021
*
BLME
(1.0mg/ml) ND 52.27±0.0212
*
BLME
(0.5mg/ml) 46.17±3.7 35.90±0.01414
*
Values are mean±SEM,*p= 0.05; ND=Not determined; SEM=Standard Error of Mean; BLME=B. lacera
methanol extract
Table 5. Inhibition (%) of amylase activities of crude methanol extract of B. lacera
Test
g
roup
Total inhibit
ion of
α
-
amylase activity
Control (DW) 0
Positive Control (50µg/ml) 79.80±0.00141
*
BLME (320µg/ml) 63.15±0.0014
*
BLME (160µg/ml) 46.49±0.00147
*
BLME (80µg/ml) 29.82±0.00147
*
DW=Distilled water, Values are mean±SEM,*p=0.05; SEM=Standard Error of Mean; BLME=B. lacera
methanol extract
4. DISCUSSION
Diarrhea results from an imbalance between the absorptive and secretory mechanisms in
the intestinal tract, which is accompanied by an excess loss of fluid in the faeces. In some
types of diarrhea, the secretory component predominates, while other types of diarrhea are
characterized by hyper motility. Castor oil causes diarrhea due to its active metabolite,
ricilonic acid [20], which stimulates the peristaltic activity in the small intestine, leading to the
changes in the electrolyte permeability of the intestinal mucosa. Its action stimulates the
release of endogenous prostaglandins [21]. The results of the present study show that there
has been a statistically significant reduction in the incident and severity of diarrhea with the
crude extract of B. lacera in experimental animals. The plant extract at 100, 200mg/kg b.w.
doses significantly lowered several typical parameters of diarrhea. Further studies are
required to confirm the underlying mechanism of the observed activity of the plants.
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1685
Results obtained from the present study revealed that the extract exhibited a relatively high
inhibitory activity against the tested fungal strains, with Blastomyces dermatitidis being the
most sensitive fungus. Amongst Gram-positive bacteria, B. cereus and Staphylococcus
aureus were the most sensitive (widest inhibition zones), whereas Shigella dysenteriae was
the most sensitive Gram-negative bacteria. The antibacterial activity may be due to the
presence of flavone type compounds in this plant [22].
The adrenergic and dopaminergic system have been shown to play a role in anxiety.
Benzodiazepine have been extensively, used for the last 40 years to treat several forms of
anxiety, but due to their unwanted side effects, alternative treatment strategies were sought
with favorable side effect profiles. Medicinal plants are a good source to find new remedies
for these disorders. Despite the wide spread traditional use of B. lacera for treating various
disorders, there are no reports of scientific evaluation of its anxiolytic activity. The present
work demonstrates that B. lacera extract had anxiolytic activity in mice as evident by open
field and hole cross models [23].
The red blood cell stability test is based on the result that a number of non-steroidal anti-
inflammatory agents inhibit heat-induced rupture of erythrocytes, most probably by
stabilizing the membrane of the cell. The erythrocyte membrane may be considered as a
model of the lysosomal membrane. Agents that can prevent the rupture of the latter, and
thereby prevent damage to the tissue caused by the release of the hydrolytic enzymes
contained within the lysosome maybe expected to improve some symptoms of inflammation.
It has been demonstrated that certain herbal preparations were capable of stabilizing the red
blood cell membrane and this may be indicative of their ability to exert anti-inflammatory
activity
[24].
The anti-diabetic activity of medicinal plants could be evaluated using several methods; in
vitro α-amylase inhibitory assay is one of such techniques. The extract of B. lacera inhibited
α-amylase but its activity was significantly less than that of positive control acarbose
(p=0.05). The inhibitory effect of methanol extract was comparable to than that of the
positive control, indicating that the α-amylase inhibitory ability resides in this extract. Alpha-
amylase is an enzyme responsible for breaking down of α-1,4-glycosidic bonds in starch.
Therefore, the enzyme increases the availability of glucose in the blood. B. lacera extract
could be useful in post-prandial hyperglycemia by reducing the hydrolysis of carbohydrates.
The observed activity may be due to the presence of chemical constituents such as phenolic
compounds (tannins and flavonoids) and terpenoids in the extract [25,26]. Phenolics have
been reported to inhibit α-amylase activities. They also have anti-hyperglycemic activity and
inhibit the development of diabetes [27,28].
5. CONCLUSION
These primary findings suggest the presence of bioactive secondary metabolites in this plant
extract that are responsible for anti-diarrheal, antimicrobial, anxiolytic, anti-atherothrombosis,
membrane stabilizing and alpha-amylase inhibitory activities.
CONSENT
Not applicable.
British Journal of Pharmaceutical Research, 4(13): 1677-1687, 2014
1686
ETHICAL APPROVAL
Not applicable.
COMPETING INTERESTS
Authors have declared that no competing interests exist.
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Chapter
Q.1 Regulations for the care and use of laboratory animals in various countriesQ.2 Techniques of blood collection in laboratory animalsQ.2.1 IntroductionBlood is collected from laboratory animals for various scientific purposes, for example, to study the effects of a test drug on various constituents, such as hormones, substrates, or blood cells. In the field of pharmacokinetics and drug metabolism, blood samples are necessary for analytical determination of the drug and its metabolites. Blood is also needed for some in vitro assays using blood cells or defined plasma protein fractions.The techniques for blood collection depend on specific factors which differ from one experiment to the other. There is a difference between terminal and non-terminal blood collection techniques. The conditions of blood collection at the end of an experiment which includes death of the animal (terminal experiment) are completely different (anesthesia, volume of blood) from those of single or repeated bloo ...
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