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Pharmacological Evaluation of Lavandula stoechas L. for Ethanol-induced Gastric Mucosal Ulce

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Background: The constituents of Lavandula stoechas L. possess antioxidant properties that help in protecting the mucosal cells from oxidative damage and speed up the healing process however, its role in the treatment of ethanol-induced peptic ulcers is not clear. Objectives: We aimed to evaluate the pharmacological potential of Lavandula stoechas L. extracts for anti-ulcer activity, and compare with the standard drugs and to explore novel treatment for peptic ulcer. Methods: We evaluated anti-ulcer potential of plant extract in ethanol-induced ulcer model in rats. Omeprazole and ranitidine were standard drugs. After 5 h of disease induction, animals were sacrificed to get tissues for histological evaluation and ulcer index was measured. While the antimicrobial potential of Lavandula stoechas L. aqueous and methanolic extracts was evaluated against different bacterial stains using standard antibiotic discs. Qualitative phytochemical and GCMS analysis were performed to identify novel constituents. Results: The methanolic extract of Lavandula stoechas L. showed antimicrobial activity against Proteus Mirabilis while the GCMS based analysis revealed the presence of 10 phytochemicals including camphor (antimicrobial agent). The aqueous extract showed significant anti-ulcer activity in ethanol-induced gastric (P < 0.001) and duodenal (P < 0.01) ulcers when compared with controls. Aqueous and methanolic Lavandula stoechas L. extracts showed strong free radical scavenging activity. Conclusion: Lavandula stoechas L. extract possess antimicrobial and anti-ulcer activity in alcohol-induced ulcer model in experimental animals.t
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Pharmacological Evaluation of Lavandula stoechas L. for Ethanol-induced Gastric Mucosal Ulce
ISSN (Print): 2521-8514 ISSN (Online): 2521-8484
RADS J. Pharm. Pharm. Sci.
47
ORIGI N A L A R T I C L E
Pharmacological Evaluation of Lavandula stoechas L.
for Ethanol-induced Gastric Mucosal Ulcer
Muhammad Ahsan Javed1, Mahtab Ahmad Khan2,*, Haroon Arshad3, Nazia Aslam4, Azeem Ahmed Iqbal3,
Asif Ali5, Mulazim Hussain Bukhari6
1Faculty of Allied Health Sciences, The University of Lahore, Lahore, Pakistan,
2Faculty of Pharmacy, University of Central Punjab, Lahore, Pakistan,
3Government of the Punjab, Health Department, Lahore, Pakistan.
4Faculty of Pharmacy, The University of Lahore, Lahore, Pakistan,
5Institute of Molecular Biology, The University of Lahore, Lahore, Pakistan.
6Department of Histopathology, University College of Medicine and Dentistry, Lahore, Pakistan.
A BS TR A CT
Authors Contributions
1, 2 designed the Study. Ahsan
1 experiment and compiled the data.
7 histopathological results were descripted
3, 4 analyzed the data,
3 manuscript was written
1, 5, 6 coordinated the project,
2, 3 final version of the manuscript was
prepared
Article info.
Received: January 09, 2020
Accepted: July 30, 2020
Funding Source: Nil
Conflict of Interest: Nil
Cite this article: Javed MA, Khan MA, Arshad
H, Aslam N, Iqbal AA, Ali A, Bukhari MH.
Pharmacological Evaluation of Lavandula
stoechas L. for Ethanol-induced Gastric
Mucosal Ulce. RADS J Pharm Pharm Sci.
2020; 8(1):47-57.
*Address of Correspondence Author:
raomahtab@yahoo.com,
mahtab.ahmad@ucp.edu.pk
Background: The constituents of Lavandula stoechas L. possess antioxidant
properties that help in protecting the mucosal cells from oxidative damage
and speed up the healing process however, its role in the treatment of
ethanol-induced peptic ulcers is not clear.
Objectives: We aimed to evaluate the pharmacological potential of
Lavandula stoechas L. extracts for anti-ulcer activity, and compare with the
standard drugs and to explore novel treatment for peptic ulcer.
Methods: We evaluated anti-ulcer potential of plant extract in ethanol-
induced ulcer model in rats. Omeprazole and ranitidine were standard drugs.
After 5 h of disease induction, animals were sacrificed to get tissues for
histological evaluation and ulcer index was measured. While the antimicrobial
potential of Lavandula stoechas L. aqueous and methanolic extracts was
evaluated against different bacterial stains using standard antibiotic discs.
Qualitative phytochemical and GCMS analysis were performed to identify
novel constituents.
Results: The methanolic extract of Lavandula stoechas L. showed
antimicrobial activity against Proteus Mirabilis while the GCMS based analysis
revealed the presence of 10 phytochemicals including camphor (antimicrobial
agent). The aqueous extract showed significant anti-ulcer activity in ethanol-
induced gastric (P < 0.001) and duodenal (P < 0.01) ulcers when compared
with controls. Aqueous and methanolic Lavandula stoechas L. extracts
showed strong free radical scavenging activity.
Conclusion: Lavandula stoechas L. extract possess antimicrobial and anti-
ulcer activity in alcohol-induced ulcer model in experimental animals.
Keywords: Antimicrobial and anti-ulcer activity, Bacterial spectrum, Free
radicals, Lavandula stoechas L., Gas chromatography
Pharmacological Evaluation of Lavandula stoechas L. for Ethanol-induced Gastric Mucosal Ulce
ISSN (Print): 2521-8514 ISSN (Online): 2521-8484 48
RADS J. Pharm. Pharm. Sci.
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INT ROD UCT IO N
Gastric ulcer is a common gastrointestinal disease
that affects appoximately 5% of people globally [1].
An imbalance between the potential harmful (acid,
pepsin, bile, drugs) and the protective (mucus &
bicarbonate secretion, prostaglandins, nitric oxide)
factors is the main cause of gastric ulcer [2].
Excessive alcohol intake, caffeine, tobacco, as well
as stress may also lead to gastric ulcer [3]. The
motility and the absorption capacity of gastrointestinal
tract may be compromised upon prolong exposure to
alcohol. The severity of ulcer can be accessed on the
basis of ulcer index or score calculated as percentage
of the ratio of ulcerated area to total surface area of
the glandular stomach [4]. Pharmacological treatment
options for gastric ulcer include H2-receptor blockers
(ranitidine), proton-pump inhibitors (PPIs, i.e.
omeprazole) and mucosal protecting agents (i.e.
sucralfate) [5]. The adverse effects such as drug
allergy, collageneous colitis and interstitial nephritis
associated with commonly used anti-ulcer agents,
urge the need of new therapeutic strategies for peptic
ulcer [6].
A number of studies have been conducted on
naturally occurring medicinal plants containing
essential ingredients that may be useful ailment for
gastric ulcer [7]. Genus Lavandula belongs to
Lamiaceae family with most important species
including Lavandula stoechas L. and Spike Lavender
(Lavandula Spica L.). Francesca Algierion and her
colleagues evaluated the anti-inflammatory potential
of Lavandula stoechas L. extracts in colitis model in
rats. The results showed that Lavandula stoechas L.
hydroalcoholic extract possessed antioxidant and
intestinal anti-inflammatory activities and therefore,
can improve the healing process as well as intestinal
epithelial barrier [8].
Oral administration of ethanol to experimental animals
induced gastrointestinal lesions [9, 10]. The process
of ethanol-induced injury is not fully known however; it
causes a disturbance in gastric mucosal integrity
through exfoliation of cells and thus, may lead to
gastric mucosal bleeding [11, 12]. Neutrophils are
recruited to the site of injury lead to ROS production
that results in oxidative stress to mucosal cells and
ulcer. Therefore, the use of antioxidants might be
helpful in protecting the mucosal cells from oxidative
damage and speeding up the healing process [12].
The constituents of Lavandula stoechas L. include
camphor, linalool, linalyl acetate, 1-8 cineole and γ-
terpinene while its antioxidant potential is found to be
comparable with vitamin E as reported in previous
studies [13]. In addition, literature showed that
methanolic extract of Lavandula stoechas L. (800
mg/kg/p.o.) significantly (P < 0.001) reduced MDA
(malondialdehyde) levels while increased SOD
(superoxide dismutase) and catalase levels in
scopolamine-induced-amnesic animal model [14].
Many studies has been conducted on antioxidant
enzymes like catalase, MDA, SOD and 1,1-diphenyl-
2-picryl-hydrazyl (DPPH)) for anti-ulcer activity of
various plants [15].
To the best of our knowledge, Lavandula stoechas L.
has not been evaluated for its possible role in the
treatment of ethanol-induced peptic ulcers. Therefore,
we aimed to evaluate and compare the
pharmacological potential of Lavandula stoechas L.
extracts for anti-ulcer activity with the standard drugs
and to explore novel treatment for peptic ulcer.
MAT ERI AL AND ME THO DS
Preparation of Lavandula stoechas L. extracts
Maceration method [16] was used to prepare
Lavandula stoechas L. extracts. Lavandula stoechas
L. powder (500 grams from arial parts) was
separately added to 1 L of each ethanol and distilled
water. The mixtures were preserved at research
laboratory for 14 days and the soaked powder was
stirred and degassed, twice daily. After maceration,
we filtered the mixtures through linen cloth, followed
by filtration with Whatman filter paper (0.45µm). The
solvents were evaportaed at 50 oC using a hot-air
oven until the extract dried.
Preparation of agar plates and microbial
suspensions for antimicrobial activity
Muller Hinton Agar (Biomark® Laboratories India, lot
# 0419/0087) media was autoclaved, 20 ml was
added equally in each petry dish and then cooled at
room temperature. The cultures of bacteria were
donated by the Microbiology laboratory, The
University of Lahore. These cultures were previously
characterized as Proteus, E.coli, Klebsiella,
Micrococcus luteus, Bacillus cereus and Salmonella
typhi. Agar plates were innoculated with different
bacterial strains and antimicrobial activity was
performed using solutions (1%, 0.5%, 0.25%, &
0.125%) of dry extracts powder dissolved in Dimethyl
Sulfoxide (DMSO). The antimicrobial activity was
Pharmacological Evaluation of Lavandula stoechas L. for Ethanol-induced Gastric Mucosal Ulce
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performed by the well diffusion method [17]. Solutions
of extracts were introduced into the wells and the
agar plates were kept at 37 °C overnight in a laminar
safety hood.
Qualitative phytochemical analysis
The reagents (Wagner, Hager, Dragendorff, Mayer,
Godine) were prepared for the qualitative
phytochemical analysis of Lavandula stoechas L. [18].
Tests were performed for glycosides (Borntrager &
Keller kiliani test) [19], alkaloids [20], tannins [21],
flavonoids [20], saponins [21], carbohydrates [21],
proteins and terpenoids [21] by following previously
published methods.
GCMS analysis
We used GCMS based method for qualitative
analysis of phytochemicals present in ethanolic and
water extracts of Lavandula stoechas L. [22]. The
sample was washed with de-ionized water, shade-
dried for 10 days, powdered and stored in zipper
plastic bags for further processing. The dried powder
(100 g) was dipped in ethanol (1 L, 95%) for 3 days
followed by filtration using Whatmann filter paper. The
filtration process was repeated again and the filtrate
was concentrated at 40°C and the resulting extracts
were kept at 4°C.
The GCMS, consisted of pumps (Agilent Technology,
USA, Model; 7890A), spectrophotometer (Agilent
Technology, USA, Model; 5975C) and column (HP-
5MS, 30 m x 250 µm x 0.25 µm, Agilent Technology,
USA). Ethanol was used as solvent and reagent in
this procedure (BDH, UK, 10107 7Y) Following
GCMS parameters were used: oven program; 60°C
for 0 minute then 10°C/minute to 300°C for 4 minutes,
run time; 29 minute, heater; on, temperature program;
280°C for 0 minute, inert gas; helium (99.99%), flow;
1 ml/minute, acquisition mode; scan, scan mass
range; 50-650, solvent delay; 4 minute, EMV mode;
relative, relative voltage; 59 and resulting EM voltage;
1306. The identification of compounds was done on
the bases of Wiley & NIST libraries and the
comparison of retention time peaks.
Experimental animals
Male albino rats were randomly grouped (5 in each
group), kept under controlled environment
(temperature 24 ± 2 °C, humidity 45-55% and 12 h
light/dark cycle) and fed with ad libitum diet [23]. All
protocols were followed according to the agreement
of the Institutional Animal Ethics Committee,
University of Lahore, Lahore, Pakistan (IREC-2018-
FEB-49).
Gastroprotective potential of Lavandula stoechas
L. extract
Solutions of omeprazole (4 mg/ml) and ranitidine (25
mg/ml) were made in distilled water. Animals were
kept on fasting overnight and orally received following
different treatments : group I and II (normal and
disease control respectively), group III; omeprazole
(20 mg/kg) [24], group IV; ranitidine (30 mg/kg) [25]
and group V; Lavandula stoechas L. extract (300
mg/kg) [26]. After 1 h, disease (gastric mucosal ulcer)
was induced in animals (group II-V) by oral
administration of 1 ml of ethanol (80%) [27]. After 5 h,
animals were anesthetized using xylazine (10 mg/kg,
intraperitoneal) & ketamine (100 mg/kg,
intraperitoneal) [28] and sacrificed to obtain the
stomach. Ulcerative lesions were measured by
planimetry the ulcer area was calculated [29]. Gastric
contents were collected, stomachs were washed with
normal saline and preserved for morphological
examination. For histological examination, excised
stomachs were fixed with formalin (10 %), dehydrated
and cleared using paraffin & xylene. Then the tissues
were cut into slices (4-5 mm), stained using
hematoxylin & eosin (H & E) dyes and examined
under microscope [29].
Antioxidant activity of Lavandula stoechas L.
extract
In-vivo antioxidant potential of Lavandula stoechas L.
extract has already been reported in literature [29].
For In-vitro antioxidant activity, the extract and
ascorbic acid was separately dissolved in DMSO (1
mg/mL). DPPH solution (1 ml, 0.1 mM in DMSO) was
added to 3 ml of test samples in various
concentrations (10 - 100 µg/ml). The mixture was
incubated (30 min, room temperature) and
absorbance was recorded at 517 nm.
Statistical analysis
As the data was normally distributed with a bell
shaped histogram so, the sum results were
expressed by means ± SEM (standard error of mean).
Parametric data were assessed by one-way ANOVA
(analysis of variance) followed by Dunnett’s test.
Graphics and statistical hypothesis testing were done
using Graph Pad Prism version 5.0 and IBM SPSS
version 19. Values P < 0.05 were considered as
statistically significant, P < 0.01 were considered as
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very significant and those with P < 0.001 were
regarded as highly significant.
RES ULT S
Antimicrobial activity with ethanolic extract
Ethanolic extract (1%) of Lavandula stoechas L.
showed antimicrobial activity against Proteus mirabilis
as evident by 3 mm inhibitory zone using augmentin
(amoxicillin 20 µg + clavulanic acid 10 µg) as
standard disk (Figure 1a, Table 1). The ethanolic
extract did not show any antimicrobial activity against
M. luteus and E.coli (using gentamycin 10 µg as
standard disk) (Fig 1b-c). No antimicrobial activity
was found against Klebsiella using augmentin
(amoxicillin 20µg + clavulanic acid 10 µg) as standard
disk (Fig. 1d) and against B. cereus & S. typhi using
Ciprofloxacin 5 µg as standard disk (Fig. 1e-f).
Antimicrobial activity of Lavandula stoechas L.
aqueous extract was determined against different
bacterial species using standard disks in same
experimental settings as used for ethanolic extracts
(Fig. 2a-f). No antimicrobial activity was seen against
any of the bacterial species used in the experiment
(Fig. 2a-f).
Table 1. Antimicrobial activity of Lavandula stoechas L. ethanolic and water extracts at 1%, 0.5%, 0.25%
and 0.125% dilutions in Dimethyl Sulfoxide (DMSO).
Microorganism
Ethanolic extract
Aqueous extract
Crude
Extract
1%
0.5%
0.25%
0.125%
Crude
Extract
1%
0.5%
0.25%
0.125%
Proteus
+ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
M. Luteus
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
E. coli
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
Klebsiella
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
B. Cereus
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
S. typhi
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve Antimicrobial activity is present, -ve Antimicrobial activity is not present
Figure 1. Antimicrobial activity of ethanolic extract from Lavandula stoechas L. against different bacterial
species: a; using augmentin, antimicrobial activity was found positive against Proteus Mirabilis, b & c; using
gentamycin, no antimicrobial activity was found positive against M. Luteus and E.coli, d; using augmentin, no
antimicrobial activity was found positive against Klebsella, e & f; using ciprofloxacin, no antimicrobial activity
was found positive against B. Cereus and Salmonella Typhi.
Pharmacological Evaluation of Lavandula stoechas L. for Ethanol-induced Gastric Mucosal Ulce
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Figure 2. Antimicrobial activity of Lavandula stoechas L. aqueous extract against different bacterial species: a;
using augmentin, no antimicrobial activity was found positive against Proteus Mirabilis, b & c; using gentamycin,
no antimicrobial activity was found positive against M. Luteus and E.coli, d; using augmentin, no antimicrobial
activity was found positive against Klebsella, e & f; using ciprofloxacin, no antimicrobial activity was found
positive against B. Cereus and Salmonella Typhi.
Table 2. Phytochemicals identified in GCMS analysis.
Sr #
Retention time
(min)
Identified compound
Mol. formula
Mol. wt
1
6.39
Cyclohexanone,2-methyl-5-(1methylethenyl) also known as
camphor
C10H16O
152
2
11.025
4,7,10-Hexadecatrienoic acid, methyl ester
C17H28O2
264
3
12.247
9-octadecenoic acid , 2-hydroxy-3-[(1-
oxooctadecyl)oxy]propyl ester
C39H74O5
622
4
12.498
9-octadecenoic acid-2-(9-octadecenyloxy)ethyl ester
C38H72O3
576
5
13.170
6,9,12,15-docosatetraenoic acid, methyl ester
C23H38O2
346
6
16.177
2,4,6,8,10-tetradecapentaenoic acid,9a-acetoxy-
1a,1b,4,4a,5,7a,7b,8,9,9a-decahydro-4a,7b-dihydroxy-3-
(hydroxymethyl)-1,1,6,8tetramethyl-5-oxo-
1Hcyclopropa[3,4]benz[1,2-e]azulen-9yl ester(limonoic acid)
C36H46O8
606
7
19.429
9,12-octadecadienoic acid, methyl ester(methyl linoleate)
C19H34O2
294
8
19.538
10-octadecenoic acid, methyl ester(methyl octadec-9-enoate)
C19H36O2
296
9
19.959
Heptadecanoic acid,16-methyl-,methyl ester(methyl stearate)
C19H38O2
298
10
20.991
9-octadecenoic acid, 2-hydroxy-1,3-propanediyl ester
C39H72O5
620
Qualitative phytochemical analysis of Lavandula
stoechas L.
The results of chemical tests of Lavandula stoechas
L. ethanolic extracts revealed that it contained
saponins, glycosides, phenols, tannins and
terpenoids. The results of GCMS based qualitative
analysis showed that 10 different phytochemicals
were identified in the ethanolic extract as summarized
in Table 2. Fig. 3 depicts the GCMS spectrum of
phytochemicals identified in the qualitative analysis.
Of the 10 identified molecules, one was
cyclohexanone, 2-methyl-5-(1methylethenyl) which is
commonly known as camphor.
Pharmacological Evaluation of Lavandula stoechas L. for Ethanol-induced Gastric Mucosal Ulce
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Figure 3. GCMS spectrum of phytochemicals identified in the qualitative analysis. The retension time mentioned
on each peak corresponds to one phytpchemical.
Figure 4. Effects of Lavandula stoechas L. aqueous extract on ulcer index: a; gastric ulcers, b; duodenal ulcers
in ethanol-induced ulcer model of experimental rats. Values are expressed as Means ± SEM, n=5; One way
ANOVA followed by Dunnett’s multiple comparison test was used; *** = p < 0.001 (highly significant results); ** =
p < 0.01 (very significant) & * = p < 0.05 (significant).
Gastroprotective effects of Lavandula stoechas L.
extract
Oral administration of ethanol successfully induced
the peptic ulcer in animals. Oral administration of
Lavandula stoechas L. aqueous extract caused a
significant reduction in ulcer index for both gastric (P
< 0.001) and duodenal ulcers (P < 0.01) as compared
to disease control as shown in Fig. 4a-b.
Administration of standard drugs; omeprazole and
ranitidine also caused marked reduction in gastric and
duodenal ulcers (P < 0.05).
The histological examination of rat stomach and
duodenum revealed that the intraperitoneal
administration of ethanol caused disruption in the
mucosal epithelium as shown Fig. 5c-d. Oral
administration of plant extract markedly improved the
mucosal integrity both in animal stomach and
duodenum (Fig. 5e-f). Moreover the standard drugs;
omeprazole and ranitidine also caused significant
improvement in gastrointestinal epithelial structure as
depicted in Fig. 5i-j.
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Figure 5. Histological examination of stomach and
duodenum respectively from: a-b; Normal control
group, c-d; Ethanol treated rats resulting in disruption
of epithelium, e-f; Omeprazole treated rats resulting in
normalization of epithelium, g-h; Ranitidine treated
rats resulting in normalization of epithelium, i-j; Rats
treated with Lavandula stoechas L. aqueous extract
showing complete normalization of epithelial
architecture.
Figure 6. Showing mean % inhibition of ascorbic
acid and Lavandula stoechas L. aqueous &
methanolic extracts by using DPPH method at
various concentrations. X-axis shows different
concentration (µg/mL) and y-axis shows %
inhibition. Values are expressed as Means ± SEM;
n=5.
In-vitro antioxidant activity of Lavandula stoechas
L. extract
In-vitro free radical scavenging activity of Lavandula
stoechas L. aqueous and methanolic extracts was
determined in context of their ability to reduce the
DPPH, a stable free radical using ascorbic acid as
reference standard. Both the aqueous and methanolic
extracts showed strong concentration dependent
DPPH reducing activity (Fig. 6).
DIS CUS S ION
Peptic ulcer disease is characterized by the erosion of
gastric and duodenal mucosa. It affects 5% of the
world population with continuously increasing disease
incidence every year. The etiology of the peptic ulcer
disease is not completely understood however;
alcohol, smoking, stress and nutritional deficiencies
are major contributors to the disease [30]. The use of
H2 receptor antagonist (H2RAs) and PPIs have
significantly decreased the incidence of ulcerative
complications [31] however; both may cause
adverse effects (drug allergy, collageneous colitis and
interstitial nephritis) [32]. A number of studies have
been conducted on naturally occurring medicinal
plants containing essential ingredients that can be
used in the treatment of peptic ulcer. Lavandula
stoechas L. is a medicinal plant and its hydroalcoholic
extract possessed antioxidant and intestinal anti-
inflammatory activity thus, can improve the healing
process as well as intestinal epithelial barrier [33].
The present study was conducted to investigate the
anti-ulcer potential of Lavandula stoechas L. extracts
Pharmacological Evaluation of Lavandula stoechas L. for Ethanol-induced Gastric Mucosal Ulce
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in animal model of ethanol-induced peptic ulcer and
to explore novel treatment strategies for peptic ulcer.
Phytochemical analysis, antimicrobial and free radical
scavenging activity of Lavandula stoechas L. extracts
were also evaluated.
Antimicrobial activity of Lavandula stoechas L. extract
against various periodontal bacterial strains was
evaluated by well diffusion method. The ethanolic
extract of Lavandula stoechas L. showed
antimicrobial activity against Proteus mirabilis as
confirmed by (3 mm) zone of inhibition while it was
completely ineffective against B. cereus, E.coli,
Klebsiella, M. luteus, and S. typhi (Fig. 1). Moreover,
the water extract of the Lavandula stoechas L. did not
show any antimicrobial activity against any of the
bacterial species used (Fig. 2). The results of
antimicrobial activity, obtained in the present study,
were consistent with existing literature for instance
the bacteriostatic activity of essential oil of the
Lavandula stoechas L. has already been reported
against periodontal bacteria [34].
The phytochemical analysis of Lavandula stoechas L.
ethanolic extract performed in the present study
revealed that it contains saponins, glycosides,
steroids, phenols, tannins, and terpenoids (Table 1).
In order to investigate the possible phytochemicals
responsible for antimicrobial activity of ethanolic
extract, GCMS based assay was performed which
showed that ethanolic extract contained 10 different
novel constituents (Table 2). Of these 10 constituents,
one was cyclohexanone,2-methyl-5-(1methylethenyl)
which is commonly known as camphor. The results of
phytochemical analysis correlated well with previously
reported studies such as gas chromatographic
analysis of Lavandula stoechas L. extract revealed
the presence linalyl acetate, γ-terpinene and camphor
[35]. According to literature, antipruritic and
antimicrobial activity of camphor has been well
documented. So, it is conceivable the antimicrobial
activity of Lavandula stoechas L. extract against
Proteus mirabilis might be due to camphor.
In the present investigation, oral administration of
ethanol successfully induced the peptic ulcer in
experimental rats [35]. However, oral treatment of rats
with Lavandula stoechas L. prior to induction of
disease protected the animals against both ethanol-
induced gastric (P < 0.001) and duodenal (P < 0.01)
ulcers as compared to disease control (Fig. 4a-b). In
addition, the standard drugs; i.e. omeprazole and
ranitidine also protected the animals against peptic
ulcer. The exact mechanism how ethanol causes
gastric lesions is not fully understood [9, 10] however,
ethanol is reported to produce a disturbance in gastric
mucosal barrier via exfoliation of cells [11, 35]
followed by the recruitment of inflammatory cells to
the site of injury and production of reactive oxygen
species (ROS). As ROS and other mediators of
inflammation may lead to oxidative damage [12, 36]
therefore, by scavenging free radicals, antioxidants
might prove as useful gastroprotective agents. In the
present study, both the aqueous and methanolic
Lavandula stoechas L. extracts showed strong free
radical scavenging activity as determined by DPPH
method (Fig. 6) which was comparable with that of
ascorbic acid (reference standard). It is conceivable
that the gastroprotective effects of Lavandula
stoechas L. aqueous extract were due to its free
radical scavenging activity.
The above discussed gastroprotective effects were
also supported by the histological examination of
stomach and duodenum derived from animals.
Ethanol treated rats showed necrosis of gastric
epithelium, manifested by disappearance of nuclei
and aggregation of inflammatory cells as compared to
control group, that might be due to the formation of
free radicals and oxidative stress induced by ethanol
(Fig. 5c-d) and this was quite consistent with the
available literature [37]. In addition, treatment of the
animals with Lavandula stoechas L. aqueous extract
lead to restoration of gastric epithelium integrity (Fig.
5i-j) which further confirmed the gastroprotective
effects which were comparable with standard drugs
(omeprazole & ranitidine). Our results were also
comparable with previous reports of other natural
gastroprotective agents i.e. Acanthus ilicifolius,
Anogeissus latifolia, Berberis vulgaris & Argyreia
speciose etc [38]. Based upon the findings of the
present investigation, it is quite evident that
Lavandula stoechas L. extract possessed
antimicrobial activity against Proteus mirabilis and
also proved effective against ethanol-induced ulcers
in animal model.
CON CL U SI ON
It is concluded from the study that Lavandula
stoechas L. methanolic extract posses antimicrobial
activity against periodontal bacteria like Proteus
mirabilis which might be due to the presence of
camphor as confirmed by the GCMS based
Pharmacological Evaluation of Lavandula stoechas L. for Ethanol-induced Gastric Mucosal Ulce
ISSN (Print): 2521-8514 ISSN (Online): 2521-8484 55
RADS J. Pharm. Pharm. Sci.
55
phytochemical analysis. The aqueous extract of
Lavandula stoechas L. also showed gastroprotective
potential in ethanol-induced gastric and duodenal
ulcer model of experimental animals as confirmed by
histomorphological changes, those might be due to its
strong free radical scavenging activity. In traditional
medicine, bacterial spectrum is a step towards the
utilization of plant extracts against specific bacterial
infections. So, Lavandula stoechas L. containing
pharmaceutical dosage forms need to be developed
and tested against Proteus infection and peptic ulcer
disease in animal models. Moreover, the efficacy of
Lavandula stoechas L. against other bacterial strains
still needs to be tested in future studies.
LIM ITA TIO N
Due to the limited financial resources and
geographical area, the findings of this study could not
be generalized. Therefore, there is a need to validate
these findings on a larger scale before entering into
preclinical trials. The literature available about other
phytochemicals, identified in the GCMS spectrum, is
very limited so, these compounds need further testing
and evaluation.
ACK NO W LED GEM ENT
We are greatly thankful to Dr. Farheen Ansari,
Ph.D Microbiology, Assistant Prof. for platform
support at Institute of Molecular Biology. We are
grateful to Ms. Naureen Zahra M. Phil
Microbiology, Lecturer, Institute of Molecular
Biology for technical expertise for the preparation
of agar plates. We also thank Mr. Ibrar Hussain
Lab Assistant, Microbiology, Institute of Molecular
Biology. We are also thankful to Ali Imran,
Department of Pharmacy and Adnan Yaqoob,
Lahore School of Nursing, University of Lahore for
technical assistance.
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the original work is properly cited.
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