Shirisha Bongu / Vol 2 / Issue 2 / 2012 / 82-87.

e-ISSN 2249 - 7749

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International Journal of

Pharmacological Screening Methods

www.ijpsmjournal.com

ANIMAL MODELS IN EXPERIMENTAL GASTRIC ULCER

SCREENING-A REVIEW
Shirisha Bongu1*, Subash Vijayakumar2
1

Department of Pharmacology, Vaagdevi College of Pharmacy, Hanamkonda, Warangal, Andhra Pradesh, India.
Department of Pharmacy Practice, Vaagdevi College of Pharmacy, MGM Hospital, Warangal, Andhra Pradesh, India.

ABSTRACT
Gastric ulcer is a very common global problem now days. Among various causes of gastric ulceration lesions caused
by stress, alcohol consumption, H.pylori infection use of NSAIDs, Uneven food habits, Gastric irritants. Hyper secretion of
gastric acid is a pathological condition, which occurs due to uncontrolled secretion of hydrochloric acid from the parietal cells
of gastric mucosa through the proton pumping H+ K+ ATPase. The modern approach to control gastric ulcer is to inhibit
gastric acid secretion, to promote gastro protection.
key words: Gastric ulcer, Stress, NSAIDs, Gastric acid.
INTRODUCTION
Ulcers are the areas of degeneration and necrosis
of gastro intestinal mucosa exposed to acid of the
alimentary tract that is exposed to hydrochloric acid and
pepsin they occur most commonly (98-99%) in either the
duodenum or the stomach in the ratio 4:1 [1]. Ulcers can
occur in the stomach, where they are called gastric ulcers
or they can occur in the first portion of the intestine called
as duodenal ulcers. "Peptic Ulcer" is the term used to
describe either or both of these two types of ulcer [2].
Ulcers cause gnawing, burning pain in the upper
abdomen. These symptoms frequently occur several hours
following a meal, after the food leaves the stomach but
while acid production is still high. Instead of pain, some
patients experience intense hunger or bloating. Other
patients have no pain but have black stools, indicating that
the ulcer is bleeding. Bleeding is a very serious
complication of ulcers [2].
Ulcers occurs due to imbalance between the
offensive (gastric acid secretion ) and defensive (gastric
mucosal integrity) factors. The aggressive and protective
factors in the stomach are acid pepsin secretion, mucosal

barrier, blood flow, cellular regeneration, prostaglandins

and epidermal growth factors. Sometimes the gastric
mucosa is continuously exposed to potentially injurious
agents such as pepsin, bile acids, food ingredients,
bacterial products (H.pylori) and drugs. Factors such as
stress, smoking, nutritional deficiency and ingestion of
NSAIDS all can increase the incidence of gastric ulcers. It
is reported that prolonged anxiety, emotional stress,
haemorrhagic surgical shock, burns and trauma are known
to cause severe gastric irritations [3].
THE REGULATION OF ACID SECRETION BY
PARIETAL CELLS
The regulation of acid secretion by parietal cells is
especially important in the pathogenesis of peptic ulcer,
and constitutes a particular target for drug action. The
secretion of the parietal cells is an isotonic solution of HCl
(150 m mol/l) with a pH less than 1, the concentration of
hydrogen ions being more than a million times higher than
that of the plasma. The Cl- is actively transported into
canaliculi in the cells that communicate with the lumen of
the gastric glands and thus with the stomach itself. This Cl -

Corresponding Author:- Shirisha Bongu Email:- shirisha46@gmail.com

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secretion is accompanied by K+, which is then exchanged

for H+ from within the cell by a K+/H+ ATPase (+ and
bicarbonate ions. The later exchanges across the basal
membrane of the parietal cell for Cl-. The principal stimuli
acting on the parietal cells are:
Gastrin (a stimulatory hormone)
Acetylcholine (a stimulatory neurotransmitter)
Histamine (a stimulatory local hormone)
Prostaglandins E2 and I2 (local hormones that inhibit
acid secretion).
Gastrin
Gastrin is a peptide hormone synthesised in
endocrine cells of the mucosa of the gastric antrum and
duodenum, and secreted into the portal blood. Its main
action is stimulation of the secretion of acid by the parietal
cells. Gastrin also indirectly increases pepsinogen secretion,
stimulates blood flow and increases gastric motility.
Release of this hormone is controlled both by neuronal
transmitters and blood- borne mediators, as well as the
chemistry of the stomach contents. Amino acids and small
peptides directly stimulate the gastrin-secreting cells [4].
Acetylcholine
Acetylcholine is released from (e.g.vagal) neurons
and stimulates specific muscarinic receptors on the surface
of the parietal cells and on the surface of histaminecontaining cells .
Histamine
Within the stomach, mast cells (or histaminecontaining cells similar to mast cells) lying close to the
parietal cell release a steady basal release of histamine,
which is further increased by gastrin and acetylcholine. The
hormone acts on parietal cell H2 receptors, which are
responsive to histamine concentrations that are below the
threshold required for vascular H2 receptor activation.
Prostaglandins
Prostaglandins (mainly E2 and I2), synthesised in
the gastric mucosa mainly by cyclo-oxygenase-1, stimulate
mucus and bicarbonate secretion, decrease acid secretion
and cause vasodilatation, all of which serve to protect the
stomach against damage [5].
Drug therapy
Different groups of drugs are used in the treatment
of the ulcers those includes [6]
Proton pump inhibitors (omeprazole, lansaprazole,
pantaprazole, robeprazole)
H2
receptor
antagonists
(cimetidine,
ranitidine,famotidine,nizatidine)
Prostaglandin analogue (mesoprostol)
Acid neutralizers (antacids) magnesium trisillicate,
aluminium hydroxide, alginates.

Ulcer protectants (sucralfate, bismuth chelate)

Animal models used in the screening of anti ulcer

activity
Various screening models are used for the screening
of the anti ulcer activity .it helps to understanding the
aetiology of the ulcer and screening of anti ulcer agents.
Aspirin induced ulcers
Ethanol induced ulcers
Pylorus ligation induced ulcers
Water immersion stress induced ulcers
Indomethacin induced ulcers
Histamine induced ulcers
Reserpine induced ulcers
Serotonin induced ulcers
Acetic acid induced ulcers
Hydrochloric acid induced ulcers
Aspirin induced ulcers
Principle
Aspirin is a NSAID which inhibit the synthesis of
prostaglandins. Prostaglandins protect the gastric mucosa
by producing leukotrienes and bicarbonate ions. Aspirin
also inhibit the gastric peroxidase and may increase
mucosal hydrogen peroxide and hydroxyl ions level to
cause oxidative mucosal damage [5].
Procedure
Albino rats of either sex weighing between 150-200
gms are divided into five groups of six animals in group.
The animals are fasted for 24 hours. The test drug in
varying concentrations based on the design of the
experiment is administered orally in 2% gum acacia
solution 30 minute prior to aspirin at dose of 200 mg/kg. 4
hours later the rats are sacrificed by using anaesthetic ether
and their stomachs dissected and they were opened along
greater curvature for the determination of gastric lesions.
Ulcer index calculated by noting the number of ulcers per
animal and severity scored by observing the ulcers
microscopically with the help of 10x lens and scoring is
done as below [7]
0 - Normal stomach
0.5 - Red coloration
1 - Spot ulcers
1.5 - Haemorrhagic streaks
2 - Ulcer > 3 mm but 5 mm
3 - Ulcers > 5 mm
Calculation of ulcer Index
UI = UN + US + UP x 10-1
UI = Ulcer Index
UN = Average of number of ulcer per animal
US = Average of severity score
UP = Percentage of animal with ulcer

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And percentage protection was observed by using the

formula:

acidic medium in stomach for longer time and produces the

ulcer [10].

%protection=

Procedure
Albino Wistar rats of either sex weighing between
(150-200 gms) are divided into groups of a animal. sIn
this method albino rats are fasted in individual cages for 24
hours. Test drug or standard drug or control vehicle is
administered 30 minute prior to pyloric ligation. Under
light ether anaesthesia, the abdomen is opened and the
pylorus was ligated. The abdomen is then sutured. At the
end of 4 hours after ligation the animals are sacrificed with
excess of anaesthetic ether , and the stomach is dissected
out gastric juice is collected were drained into tubes and
were centrifuged at 1000 rpm for 10 minutes and the
volume is noted. The pH of gastric juice is recorded by pH
meter. Then the contents are subjected to analysis for free
and total acidity. The stomachs are then washed with
running water to see for ulcers in the glandular portion of
the stomach. The numbers of ulcers per stomach are noted
and severity of the ulcers scored microscopically with the
help of 10x lens.
Histopathological studies were conducted by fixing
stomach tissues in 10% formalin for 24 h. The formalin
fixed specimens are embedded in paraffin and section (35m) and stained with haematoxylin and eosin dye. The
histochemical sections are evaluated by light microscopy
[11,12].
0 = Normal stomach
0.5 = Red coloration
1 = Spot ulcers
1.5 = Haemorrhagic streaks
2 = Ulcer > 3 mm but > 5 mm
3 = ulcers > 5 mm

Ethanol induced ulcers

Principle
Alcohol causes secretion of gastric juice and
decrease mucosal resistance due to which protein content of
gastric juice is significantly increased by ethanol. This
could be leakage because of plasma protein in the gastric
juice with weakening of mucosal resistance barrier of
gastric mucosa, this leading to peptic ulcer [8].
Procedure
Albino rats of either sex weighing between (150-200
gms) are divided into group. The animals are fasted for 24
hours with free access water. Animals are given test drugs
or standard drug. 1 hour later 1ml/200gm of 99.80%
alcohol is administered orally to each animal. The animals
were anaesthetized 1 h latter with ether and stomach was
incised along the greater curvature and ulceration was
scored. The number of ulcers and the length of each ulcer
were measured. Ulcer index was calculated using severity
scores and avg no of ulcers per animal. Severity scores as
below [9]
0 - Normal stomach
0.5 - Red coloration
1 - Spot ulcers
1.5 - Haemorrhagic streaks
2 - Ulcer > 3 mm but 5 mm
3 - ulcers > 5 mm
Calculation of ulcer Index
UI = UN + US + UP x 10-1
UI = Ulcer Index
UN = Average of number of ulcer per animal
US = Average of severity score
UP- Percentage of animal with ulcer
Percentage inhibition was calculated by the formula
X 100
Histopathological studies were conducted by
fixing stomach tissues in 10% formalin for 24 h. The
formalin fixed specimens are embedded in paraffin and
section (3-5m) and stained with haematoxylin and eosin
dye. The histochemical sections are evaluated by light
microscopy.
Pylorus ligation induced ulcers
Principle
In Pylorus ligation pylorus part of the stomach was
ligated it helpful to produce the ulcers in rats by stopping
passage of gastric contents from stomach it creates the

Calculation of ulcer Index

UI = UN + US + UP x 10-1
UI = Ulcer Index
UN = Average of number of ulcer per animal
US = Average of severity score
UP = Percentage of animal with ulcer
%protection=
Determination of free acidity total acidity
One ml of gastric juice is pipette into 100 ml
conical flask, added 2 to 3 drops of topfers reagent and
treated with 0.01 N sodium hydroxide until all traces of red
colour disappears and the colour of the solution turns to
yellowish orange. The volume of the alkali added was
noted. This volume corresponds to free acidity. Then 2 to 3
drops of phenolphthalein solution is added and titration is
continued until a definite red tinge reappears. Again the
total volume of alkali added is noted. Acidity is calculated
by using the formula

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Acidity = volume of NaOH X Normality of

NaOHx100/0.1 m.eq/lit/100gms.
Water immersion stress induced model
Principle
Stress can arise from prolonged anxiety, tension,
and emotion, severe physical discomfort, haemorrhage and
surgical shock, burns and trauma, thereby resulting in
severe gastric ulceration.
Procedure
Stress induced ulcers were induced by force
swimming in the glass cylinder (height 45cm diameter
35cm) containing water up to 35cm maintained at 35 oc for
3 hrs. Animals were fasted 24 hrs prior to the experiment
.After the drug treatment (standard/test) animals were
allowed to swim for 3hrs then animal were dissected
stomachs were removed. All stomachs were opened along
the greater curvature ulcer index and % inhibition was
calculated and Histopathological studies conducted [13].
Indomethacin induced ulcers
Principle
Indomethacin is a non steroidal anti inflammatory
drug that inhibits the synthesis of prostaglandins which are
protective agents for gastric mucosa high doses of the
NSAIDs causes ulceration [5].
Procedure
All the animals were fasted 36 hours before
administration of Indomethacin. The animals were divided
into groups. Each rat was administered with the 20mg/kg
Indomethacin orally.30 min prior to the administration of
the Indomethacin standard/test drug was administered. The
rats were anaesthetized with ether 1 hour latter the stomach
was incised through the greater curvature and examined for
the number of lesion under the dissecting microscope by
titrating with 0.01N NaOH using phenolphthalein as an
indicator. gastric juice estimated for pepsin. Ulcer index
and % inhibition were calculated and histopathological
studies conducted for the stomach tissues [14].
Histamine induced ulcers
Principle
Histamine causes gastric ulceration by enhancing
the gastric acid secretion by directly acting on histamine
receptors of parietal cells and it also having the vasoplastic
activity [4].
Procedure
Guinea pigs weighing (300-400 gm) were divided
into groups of six each and the animals were fasted for 36
hours (water allowed) before experiment. One ml of
histamine acid phosphate (50 mg base) was administered
intraperitoneally. Promethazine hydrochloride 5 mg was
injected intraperitoneally 15 min before and 15 min after

histamine administration. The standard/test drugs were

administered by gavage 45 minutes before histamine. Four
hours after administration of histamine, the guinea pigs
were sacrificed by stunning. The anterior abdominal wall
was opened and the stomach dissected out. Stomach was
opened along the greater curvature ulcers were identified.
Severity scores were calculated. Histopathological studies
were conducted on the stomach tissues [15].
Reserpine induced ulcers
Principle
Reserpine-induced gastric ulceration has been
attributed to the degranulation of gastric mast cells and
consequent liberation of histamine which is believed to be a
cholinergically mediated [16].
Procedure
Adult albino rats weighing 150-180gms were
fasted for 24 hr. Animals were divided into different groups
following water ad libitum. Reserpine (5mg/kg)
administered intramuscularly rats. 30 min after the
administration of the standard or test drug or control
vehicle (Distilled water) intraperitoneally. All the animals
were sacrificed after 18 hr, their stomachs were removed,
opened along the greater curvature and sum of lengths
(mm) of all lesions for each rat was used as ulcer index and
percentage protection of ulcers were calculated.
Histopathological studies were performed on the stomachs
tissues [17].
Serotonin induced ulcers
Principle
Serotonin-induced gastric ulceration is believed to
arise from a disturbance of gastric mucosal microcirculation
.The development of ulcers by serotonin usually takes
about 18 hr10.
Procedure
Rats weighing 120-150gms were taken and they
were randomly divided into groups. Animals kept fasting
for 24hrs and water withdrawn 2hr before the experiment.
Serotonin creatinine sulphate (20mg/kg) was administered
subcutaneously to rats. The standard drug/test drug/ control
vehicle (Distilled water) was administered intraperitoneally
after 30 min prior to the serotonin injection. The animals
were sacrificed after 18 hr, their stomachs were removed
and opened along the greater curvature, and the ulcer index
was determined. Percentage protection of the ulcers
calculated. Histopathological studies were conducted
[18,19].
Acetic acid induced ulcers
Principle
Acetic acid enhances the ulceration in stomach by
increasing the acidity of stomach contents and acetic acid
also causes gastric obstruction leads to the ulceration.

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Procedure
Rats were anaesthetized with pentobarbitone (35
mg/kg, ip). The abdomen was opened and the stomach was
visualized. A cylindrical glass tube (6 mm in diameter) was
tightly placed upon the anterior serosal surface of the
glandular portion of stomach 1 cm away from the pyloric
end. 50% acetic acid (0.06 ml/animal) was instilled into the
tube and allowed to remain for 60 sec on the gastric wall .
After removal of acid solution, the abdomen was closed in
two layers and animals were caged and fed normally.
Standard and test drug administered orally 4 h after the
application of acetic acid and continued up to 9 days after
induction of ulcer. The animals were then sacrificed after
18 h of the last dose of drug on 10th day of experiment to
assess the ulcer size and healing. Ulcer index was
calculated based upon the product of length and width
(mm2/ rat) of ulcers [20].
Hydrochloric acid induced ulcers
Principle
Hydrochloric acid induces the ulcer by enhancing
the acidity of the stomach contents.
Procedure

Rats weighing 150-180gms are taken and they

were divided into groups. Thirty minutes after the test or
reference drug or the control vehicle treatment, 0.6 M HCl
was orally administered to each rat. After 1 h the rats were
anaesthetised with excess of anaesthetic ether and stomach
was cut open along the greater curvature, cleared of
residual matter with saline and the inner surface was
examined for ulceration. Ulcer index and % ulcer
protection. Histopathological were conducted by Gastric
tissue samples from each group were fixed in 10% formalin
for 24 h. The formalin fixed specimens are embedded in
paraffin and section (3-5m) and stained with haematoxylin
and eosin dye. The histochemical sections are evaluated by
light microscopy [21].
CONCLUSION
Several methods are there to evaluate the anti
ulcerogenic activity of the synthetic and natural products
like aspirin induced ulcers, alcohol induced ulcers, pyloric
ligation induced ulcers, indomethacin induced ulcers,
histamine induced ulcer, reserpine induced ulcers, serotonin
induced ulcer, acetic acid induced ulcers, Hydrochloric acid
induced ulcers. Most of the researcher could evaluate the
efficacy of various anti ulcer drugs by these methods.

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