Research Gate: Pharmaceutical Sciences, 1 (2013) 76-79
Antiulcerogenic Effects of Terminalia chebula (Retz.) in Pylorus Ligated Rats 1Department of Pharmaceutical Sciences, Doctor Hari Singh Gour Vishwavidyalaya Sagar-(M.P.) India2MES College of Pharmacy Sonai (M.H.) IndiaA R T I C L E I N F O A B S T R A C T Keywords:
The cause of ulceration in patients is mainly due to hyper secretion of gastric juice and also due
to hyper secretion of pepsin. In traditional system of medicine a number of herbal preparations
have been used for the treatment of peptic ulcers. The marketed poly-herbal formulation and
many synthetic drugs has been used for the treatment of gastric-ulcer diseases. In view of this, in
present study we have to evaluate the anti-ulcer activity of Terminalia chebula. Study wascarried out, on pyloric ligation induced ulcers in rats pretreated with the doses of 1ml/kgAbsolute ethanol, Extracts and fraction of Terminalia chebula, Omeprazole: 20 mg/kg body wt. After 4 hour in pyloric ligation method induced method rat was sacrificed and stomach wasremoved for observation of various biochemical and histo-pathological parameters such as pH ofgastric content, free acidity, total acidity and ulcer index were determined. The results ofpharmacological activity led to the conclusion that the Alcoholic extract and Ethyl acetatefraction of alcoholic extract exhibited activity that is more significant. Ethanol extract and its
Article History:
Ethyl acetate fraction significantly raised the pH of gastric content and thus reducing the level of
acidity in the stomach. It lowered the free and total acidity and ulcer index as compared to
control group. The antigastric-ulcer activity may be due to Gallic acid as the fraction contains
Introduction
effects and limitations Herbal medicine deals with plants
The acid-peptic diseases are those disorders in which
and plant extracts in treating diseases. These are
gastric acid and pepsin are necessary, but usually not
considered safer because of the natural ingredients with
sufficient, pathogenic factors. While inherently caustic,
no side effects. Gastric ulcer, one of the most widespread,
acid and pepsin in the stomach normally do not produce
is believed to be due to an imbalance between aggressive
damage or symptoms because of intrinsic defense
and protective factors.2 The gastric mucosa is
mechanisms. Barriers to the reflux of gastric contents into
continuously exposed to potentially injurious agents such
the esophagus comprise the primary esophageal defense.
as acid, pepsin, bile acids, food ingredients, bacterial
If these protective barriers fail and reflux occurs,
products (Helico bacter pylori) and drugs.3
dyspepsia and/or erosive esophagitis may result. 1 Ulcers
These agents have been implicated in the pathogenesis of
are sores on the lining of the digestive tract which consists
gastric ulcer, including enhanced gastric acid and pepsin
of the esophagus, stomach, duodenum and intestines. Most
secretion, inhibition of prostaglandin synthesis and cell
ulcers are located in the duodenum and they are called
proliferation growth, diminished gastric blood flow and
duodenal ulcers. Ulcers located in the stomach are called
gastric ulcers. Ulcers in the esophagus are called
Terminalia chebula (Family: Combretaceae) was one of
the traditional medicine used in many folk claims and it is
Stomach ulcer (peptic ulcer) is a breaking in the tissue
called as “King of medicine”. It is an middle-sized tree
lining of the stomach. Most are caused by infection with
leaves are ovate, or elliptic, flowers are yellowish white,
the Helicobacter pylori bacterium and not by stress or
fruits are yellowish brown in colour distributed
poor diet. Symptoms may include abdominal pain just
throughout in India.5 The plant has extensively used in
below the ribcage, indigestion, nausea, loss of appetite,
ayurveda and sidda for Constipation, diarrhea, ulcers,
vomiting, weight loss, blood in the vomit or bowel motions
gastroenteritis, asthma, cough, dyspnea, dyspepsia,
and symptoms of anaemia, such as light-headedness. The
hemorrhoids, candidiasis, parasites, malabsorption
existing treatment options include antibiotics and acid-
syndrome, hepatomegaly, vesicular and renal calculi,
urinary discharges, tumors, skin diseases, leprosy,
Although a number of antiulcer drugs such as H
antagonists, proton pump inhibitors and cytoprotectants
neuropathy, paralysis, memory loss, epilepsy, depression,
are available for ulceration all these drugs have side
diabetes, cardiovascular diseases, anorexia, wounds.
* Corresponding author: [email protected]
Research Gate: Pharmaceutical Sciences, 1 (2013)
A peptic ulcer, also known as ulcus pepticum or peptic
Pylorus Ligation induced gastric ulcers
ulcer disease, is an ulcer (defined as mucosal erosions
equal to or greater than 0.5 cm) of an area of the
Albino rats were divided into five groups of six animals
gastrointestinal tract that is usual y acidic and thus
each and fasted for 48h with free access to water. Pyloric
extremely painful. 70-90% of ulcers are associated with
ligation was performed under light ether anesthesia to
Helicobacter pylori, a spiral-shaped bacterium that lives in
each animal. Animals were given 1% Tween 80 solution or
the acidic environment of the stomach. Ulcers can also be
extracts 250mg/ kg or 20 mg/ kg omeprazole orally
caused or worsened by drugs such as aspirin and other
immediately after pylorus ligation. Animals were
sacrificed 4 h later. The stomach was carefully removed
and gastric contents were collected. The excised stomach
Therapies are directed at decreasing gastric acidity,
was studied histologically and the gastric juice was
enhancing the lower esophageal sphincter, or stimulating
centrifuged at 3000 rpm for 30 min and the supernatant
esophageal motility. In the stomach, mucus and
was used for analysis of various biochemical and
bicarbonate, stimulated by the local generation of
prostaglandins, protect the gastric mucosa. If these
defenses are disrupted, a gastric or duodenal ulcer may
Biochemical Parameters:
form. The treatment and prevention of these acid-related
Estimation of pH of gastric contents
disorders are accomplished either by decreasing the level
of gastric acidity or by enhancing mucosal protection.
The pH of Gastric contents were measured by pH meter,
Materials and Methods Table 1: pH Study of Rat Gastric juice treated with Plant Material
extracts/fraction of fruit of Terminalia chebula
Fruits of Terminalia chebula were purchased from local
Alcoholic Ethyl acetate
market of Sagar (M.P.) in the months of November and
Fraction
December 2011. Fruits were authenticated by Dr. P.
Tiwari (Herbarium no. Bot/1019) at the Department of
Mean ±SD (n=6 Significance * p<0.01); #std. Omeprazole
Botany Dr. H. S. Gour Vishwavidyalaya, Sagar (M.P.) and
preserved in the herbarium of the institute. Determination of free acidity and total acidity of Gastric contents Preparation of Extracts
The gastric contents were centrifuged at 1000 rpm for
Successive Solvent Extraction
10min. 1ml of supernatant was diluted with 9ml of
The powder of fruits (600g) were successively extracted
distilled water. A volume of 2ml diluted gastric juice was
with petroleum ether (60-80C), benzene, ethyl acetate,
titrated with 0.1N Sodium hydroxide run from a
chloroform, ethanol and water by soxhlet extraction
microburette using 3-4 drops of Topfer's reagent as
process. During the extraction, the test was performed for
indicator until canary yellow colour was observed. Volume
the completion of the process. The completion of the
of NaOH required was noted. This corresponds to free
extraction was confirmed by evaporating a few drops of
acidity. Further 2-3 drops of phenolphthalein was added
the extract on the watch glass and ensuring that no
and titrated with NaOH until pink colour was restored.
residue is found after evaporating the solvent.7 The marc
This gives total acidity. Free acidity and total acidity is
was dried in hot air oven below 40C before being
expressed in terms of ml of 0.1N HCl per 100 gms of
extracted with the next solvent. The solvent was
gastric contents. This is the same as mEq/lit. To obtain
recovered by distillation, and the last traces of solvent
this figure multiply the burette reading obtained from
were removed under vacuum. The extract obtained with
titration by 10 (Table 2 and 3). The total acidity is
each solvent was weighed to a constant weight and
expressed as m Eq/L by the following formula:
percentage on w/w basis was calculated.
Acidity = (Vol. of NaOH × N × 100)/0.1 mEq/L
Drugs and Chemicals Table 2: Free acidity of Rat Gastric contents of treated
Omeprazole drug sample was received from Sun Pharma
with fruit extracts/fraction of Terminalia chebula
and used during the experimental protocol. Alcoholic Alcoholic Fraction In vivo Study Animal selection-Wistar albino rats
Mean ±SD (n=6 Significance * p<0.01); #std. Omeprazole
Wistar albino rats weighing between 150 and 200 g were
Table 3: Estimation of Total acidity of Gastric contents
selected for the antiulcer studies. The animals were
acclimatized to standard laboratory conditions of
Alcoholic Alcoholic Fraction
temperature (22±30°C) and maintained on 12:12 h
light:dark cycle and distilled water. The animal care and
experimental protocol was in accordance with IAEC of
Department of Pharmaceutical sciences, Dr. H. S. Gour
Mean ±SD (n=6 Significance * p<0.01); #std. Omeprazole
Research Gate: Pharmaceutical Sciences, 1 (2013)
Histological Parameters
a known anti-ulcer drug, Ethyl acetate fraction of Alcoholic
Ulcer Index study and Percentage Inhibition
extract and alcoholic extract showed good results,
whereas Aqueous extract are less potent in decreasing
Rat stomach were excised and opened along the greater
gastric acidity. The results are tabulated in Table 2.
curvature, washed and stretched and studied for its
Estimation of total acidity of gastric content in terms
histology. The internal surface of stomach was examined
of ml of 0.1N HCl / 100ml of gastric content
for the presence of lesions and the extent of the lesions
was measured. The number of the lesions along the wall of
Gastric total acidity is increased to (44.67 mEq/litre) in
stomach was recorded as ulcer index and % inhibition was
control animals. Ethyl acetate fraction (34.87 mEq/litre)
also calculated (Basile, et al., 1990). Each stomach sample
showed significant decrease in total acidity (p<0.01) as
was examined grossly and the ulcers were graded using
compared to control. The decrease in total acidity by
the following system suggested by Kunchandy et al. 1985.
alcoholic extract and Aqueous extract is 41.4 and 42.56
mEq/litre) respectively. The results are tabulated in Table
Calculation of Ulcer index (UI) and percent inhibition (% I) Determination of ulcer index
Ulcer Index (UI) = (Number of ulcers in control- Number
The ulcer index in control animals was (5.18). Ethyl
acetate Fraction (3.85) significantly reduced the ulcer
Percent Inhibition (% I) =(UI of control – UI of test *100)/
index (p<0.01) as compared to control. The reduction in
ulcer index by alcoholic extract and aqueous extract is 4.4
and 4.85 respectively. Omeprazole, a standard anti-ulcer
Statistics -
drug showed ulcer index 2.19. The results are tabulated in
The ulcer index and percent ulcer inhibition and Statistical
analysis was performed by one-way analysis of variance
Table 4: Ulcer Index and ulcer Inhibition percentage
(ANOVA) followed by Dunnett’s t-test for multiple
profile of fruit extracts/fraction of Terminalia chebula
comparisons. The significance of difference was accepted
Parameters Alcoholic Alcoholic
at p < 0.05. An analysis was performed using the software
Fraction Observation
The prepared slides were viewed under light microscope
#std. Omeprazole
(100X). The observed stomach epithelium was Red
colored and severely eroded with perforations in case of
Conclusion
control group of animals. The number of ulcers was more
The pathogenesis of gastric ulcers remains still unclear,
in control group as compared to the treated groups. The
ranges from stressful lifestyle, alcohol consumption, use of
number of ulcers and severity of ulceration was less in
steroidal and non-steroidal anti inflammatory drugs
groups treated with extracts and Ethyl acetate fraction.
(NSAIDs) and drugs which stimulate gastric acid and
The presence of few ulcer sites and ulceration of moderate
pepsin secretion, Helicobacter pylori infections, smoking,
or negligible nature in case of Ethyl acetate fraction
lower socio-economic status and family history. All these
treated groups of animals gives significant indication of
represent significant risk factors that may contribute to
potent ulcer protective effects compared to control
increasing gastric damage. Now days, various approaches
have been made to study herbal drugs for treatment of
various gastrointestinal disorders. From the present
study, it was concluded that all extracts possesses
Estimation of pH of gastric content
significant anti-ulcerogenic effect at varied level, thus a
In control animals, without any drug the mean is 2.51.
specific chemical entity which is responsible for the anti-
Ethyl acetate Fraction showed significant rise in pH (3.37)
ulcerogenic effect is yet to be explored and suggested by
as compared to control. The rise in pH shown by Alcoholic
the authors at molecular level by altering protocols at
and aqueous extracts are 2.71 and 2.62 respectively.
Omeprazole, a standard drug raised the pH to 3.86, which
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Research Gate: Pharmaceutical Sciences, 1 (2013)
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Experimental Hyperlipidemia Causes an Increase in theDalia A. Hamdy, MSc and Dion R. Brocks, PhDbe caused by several factors such as genetic makeup orAbstract: To assess the influence of hyperlipidemia (HL) onsecondarily to other factors including diet and lifestyle habitsamiodarone (AM) effect in the heart, rats were pretreated with either 1and diseases or drugs.1–3 Plasma lipoproteins, w
Research Gate: Pharmaceutical Sciences, 1 (2013) 76-79
Antiulcerogenic Effects of Terminalia chebula (Retz.) in Pylorus Ligated Rats