Study of
Antiulcer Activity of Leaves of Sesbania grandiflora Linn. (Fabaceae)
Himgauri V. Naik1,
Navanath Chavan2, Himanshu
A. Deshmukh2, Pratip K. Chaskar3*,
Nikhil S. More4
Lokseva College of Pharmacy, Pune,
Maharashtra, India.
2Sanjivani College of
Pharmaceutical Education and Research, Kopargaon,
Maharashtra, India.
3Yadavrao Tasgaonkar
Institute of Pharmacy, Bhivpuri Road, Maharashtra,
India.
4Sinhgad College of Pharmacy,
Vadgaon (Bk.), Pune,
Maharashtra, India.
ABSTRACT:
Plant
extracts have been used for centuries, as popular remedies against several
health disorders. In traditional books, it was mentioned that Sesbania grandiflora
L. leaves has antiulcer activity. With reference to these, we had taken this
plant for its perfection study. The present study revealed the antiulcer
activity batches using petroleum ether, chloroform and hydroalcoholic
solvents of Sesbania grandiflora
L. The above activities observed may be due to the presence of phytochemical constituents such as alkaloids, flavonoids, triterpenes,
steroids, glycosides and tannins. In short, this project was a successful
attempt to discover new antiulcerative property from
plant source. The ethanolic extract of Sesbania grandiflora
L. prevented acute gastric injury in rats. The extracts of Sesbania grandiflora L. were subjected to
qualitative analysis for the various phytoconstituents
like alkaloids, carbohydrates, glycosides, phytosterols,
saponins, tannins, proteins, amino acids and flavonoids.
KEYWORDS: Antiulcer, Sesbania grandiflora L., phytochemical
constituents
INTRODUCTION:
Peptic ulcers are open sores that develop on the inside
lining of the stomach, upper small intestine or esophagus. Ulcer are mainly
differentiated into three types, viz, gastric ulcer - occurs
in the stomach, duodenal ulcer – occurs in the first part of the small
intestine (duodenum) and esophageal ulcer – occurs in the lower section of the
esophagus. It is often associated with chronic gastroesophageal
reflux disease. The most common symptom of a peptic ulcer is burning pain. This
pain is caused by the ulcer and is aggravated by stomach acid coming in contact
with the ulcerated area. Less often, ulcers may also cause severe signs or
symptoms such as vomiting of blood, which may appear red or black, dark blood
in stools or stools that are black or tarry, nausea or vomiting, unexplained
weight loss, chest pain (Harsh Mohan, 2000; Hoogerwerf
et al., 2006).
Although stress and spicy foods were once thought to be
the main causes of peptic ulcers, doctors now know that the cause of most
ulcers is the corkscrew-shaped bacterium Helicobacter
pylori (H. pylori). H. pylori lives and multiplies within
the mucous layer that covers and protects tissues that line the stomach and
small intestine. Often, H. pylori
causes no problems, but
sometimes, it can disrupt the mucous layer and inflame the
lining of the stomach or duodenum, producing an ulcer. One reason may be that
people who develop peptic ulcers already have damage to the lining of the
stomach or small intestine, making it easier for bacteria to invade and inflame
tissues. Also, the regular use of painkillers such as NSAIDs, smoking and
excessive alcohol consumption contribute to ulcer formation (Sura et al.,
2011; Page et al., 2002).
Sesbania grandiflora L. is a tree that grows rapidly,
provides light shade and is often grown as ornamental. This soft-wooded tree's
leaves are used as fodder and its flowers as food. Sesbania
grandiflora L. is planted in gardens for its
intercropping compatibility and soil-improving properties.
Plant extracts (Sura et al.,
2011) have been used for centuries, as popular remedies
against several health disorders. In recent years, large advances in chemical
and pharmacological studies have contributed to the knowledge about new
therapeutically active compounds obtained from the natural products. These
compounds can be used directly as leads for the development of new medicines or
as pharmacological tools to discover new active compounds. These can be
life-saving completely or may be used to improve the quality of life in
long-lasting diseases. Sesbania grandiflora L. is a tree that grows to 8-10 m in height. The
pink-red or white flowers of this papilionaceous
(pea-like flowered) legumes are unusually large (5-10 cm in length and about 3
cm wide before opening). This novelty may be the principal reason for Sesbania grandiflora L.
having been distributed by man throughout the tropics and subtropics.
Its bark
contains tannin and gum. Saponin is isolated from its
seeds. Sesbanimide isolated from seed is considered a
cancer inhibitor. All the parts of Sesbania
grandiflora L. are utilized for medicine in
Southeastern Asia and India including preparations derived from the roots,
bark, gum, leaves, flowers and fruit. In a number of cultures, the root is
applied as a poultice for application to inflammation and fever. Leaves are
used for epilepsy, gout, itch, leprosy, nyctalopia
and ophthalmia. The bark is considered astringent and
is utilized for the treatment of smallpox in Philippines, for the treatment of
ulcers in the mouth and alimentary canal in Java, for the treatment of thrush
and infantile disorders of the stomach and the compounded bark is applied to
scabies in Cambodia (Lakshmi, 2011; Bhalke et al.,
2010; Lakshmi et
al., 2011).
MATERIALS AND METHODS:
Materials
The leaves of Sesbania
grandiflora L. was collected from Ahmednagar area of Maharashtra, India, in the month of
October and were authenticated by the Government of
India, Ministry of Environment and Forest Botanical Survey of India. These
leaves were collected from Kopargaon and then subjected
to soxhlet extraction by using petroleum ether, ethyl
acetate and ethanol as solvent. Hydroalcoholic leaf
extracts of Sesbania grandiflora L. in albino mice (weighing up
to 30-40 g) up to the dose limit of 400 mg/kg was used. One fourth of the
maximum dose (100 mg/kg) tested for LD50 of each extract was selected to study
the antiulcer activities in different experimental models such as Indomethacin induced ulcer models in rats. In Indomethacin model, parameters like ulcer index, volume of
gastric juice, free acidity, total acidity, pH and total protein content of
gastric juice were estimated. No mortality was observed with these extracts up
to the maximum dose of 400 mg/kg.
All the chemicals and solvents were procured from the
local supplier. Soxhlet apparatus was used for
extraction. Rotary evaporator (Euiptronics; Model: Evator) was used for evaporation of solvent from extracts.
Extraction of Leaves
The leaves of Sesbania
grandiflora L. were collected, shade dried and
reduced to coarse powder in a mechanical grinder. The powdered material
obtained was then subjected to successive extraction in batches using petroleum
ether, chloroform and hydroalcoholic solvents in a Soxhlet extractor. Briefly, extraction procedure involved
packing a known amount of dried powdered plant material (25-50 g) in the Soxhlet extraction assembly using a filter paper thimble
followed by addition of solvent (200-300 mL) to make
one complete siphon. Additional solvent (200 mL) was
then poured into the thimble. Extraction assembly was heated at 70-80°C using heating mantle for 48-72 h till the completion
of extraction. Completion of extraction was confirmed by visualizing a TLC spot
of liquid coming out of siphon tube under UV. Collected extract was then
concentrated under reduced pressure (below 40°C). The different extracts obtained were evaporated at
45°C to get a semisolid mass.
The extracts, thus obtained, were subjected to phytochemical
analysis.
Phytochemical Estimations of the Extracts
The extracts of Sesbania grandiflora L. were subjected to
qualitative analysis for the various phytoconstituents
like alkaloids, carbohydrates, glycosides, phytosterols,
saponins, tannins, proteins, amino acids and flavonoids.
Fig
1. Total Ulcer Formed
Table 1. Phytochemical Estimation of
Petroleum Ether, Ethyl Acetate and Ethanolic extract
of leaves of Sesbania grandiflora
L.
|
Sr. |
Phytochemical Estimation |
Pet. Ether Extract |
Ethyl Acetate Extract |
Ethanolic Extract |
|
|
1 |
Test
for Alkaloids |
Hager's
Test |
- |
+ |
+ |
|
Mayer's
Test |
- |
+ |
+ |
||
|
Dragendroff's Test |
- |
+ |
+ |
||
|
Wagner's
Test |
- |
+ |
+ |
||
|
2 |
Test
for Carbohydrates |
Molisch's
Test |
+ |
- |
- |
|
Fehling's
Test |
+ |
- |
- |
||
|
Barfoed's
Test |
+ |
- |
- |
||
|
Benedict's
Test |
+ |
- |
- |
||
|
3 |
Test
for Phytosterols and
Triterpenoids |
Liebermann-Burchard Test |
+ |
- |
- |
|
Salkowski
Test |
+ |
- |
- |
||
|
4 |
Test
for Cardiac Glycosides |
Baljet
Test |
- |
- |
- |
|
Keller
Killani's Test |
- |
- |
- |
||
|
5 |
Test
for Anthraquinone Glycosides |
Borntrager’s Test |
+ |
- |
- |
|
5 |
Test
for Saponins |
Froth
Test |
+ |
- |
- |
|
6 |
Test
for Tannins |
Ferric
Chloride Test |
- |
- |
+ |
|
Lead
Acetate Test |
- |
- |
+ |
||
|
7 |
Test
for Proteins and
Amino Acids |
Biuret
Test |
+ |
+ |
- |
|
Millons's
Test |
+ |
+ |
- |
||
|
Ninhydrin
Test |
+ |
+ |
- |
||
|
8 |
Test
for Flavanoids |
Lead
Acetate Test |
- |
+ |
- |
|
Shinoda
Test |
- |
+ |
- |
||
Table 2. Gastroprotective Activity of
ELESG on Indomethacin Induced Ulcer Model
|
|
No.
of Animals |
Ulcer
Index |
%
Ulcer Formed |
%
Ulcer Protection |
|
Control |
6 |
87.83
± 2.928 |
-- |
-- |
|
Std.
(MIS 100µg/kg) |
6 |
16.83
± 1.138* |
19.33 |
29.56 |
|
Test
(ELESG 300mg/kg) |
6 |
25.75
± 2.032* |
80.66 |
70.43 |
Values are Mean ± S.E.M., *P<0.0001 extremely
significant, **P<0.01 very significant, ***P<0.05 significant as compared
to control group. The group treated for the control vehicle.
RESULTS:
All the procedures were performed and the results were
evaluated. Table 1 shows phytochemical estimation of
petroleum ether, ethyl acetate and ethanolic extract
of leaves of Sesbania grandiflora
L.
The values demonstrate the results shown by different
extracts of Sesbania grandiflora
L. Gastroprotective activity of ELESG on indomethacin induced ulcer model was studied and the
observations were recorded as shown in Table 2.
Total ulcer formed is depicted in Fig 1. Fig 2 shows
total ulcer protection and Fig 3 showed the photograph of the dissected stomach
for ELESG 300mg/kg.
Fig 2. Total Ulcer
Protection
Fig
3. ELESG 300mg/kg
Antiulcer
Activity
Although in most of the
cases, etiology of ulcers is unknown; but it is due to imbalance between
offensive (acid and pepsin) and defensive factors (mucin
secretion, cell proliferation, prostaglandin deficiency).
Increased acid
secretion and poor quality of mucus production renders easy degradation by the
enzyme attacks that also lead to the lesion on gastric mucosa and ultimately
the ulcers. It is postulated that dopamine imbalance is one of the several
factors for pathogenesis of gastric ulceration. Some drugs also readily induces
ulcers e.g. Indomethacin, aspirin, ethanol.
Antiulcer activity was
produced by the decrease in leakage and damage in mucosa and by increasing mucoprotein (mucosal) and mucus secretion i.e.
increased total hexose and hexsosamine.
To regain the balance, different therapeutic agents including plant extracts
are used to inhibit the gastric acid secretion or to boost the mucosal defense
mechanism by increasing mucus production, stabilization of surface epithelial
ulcer and interfere with the prostaglandin synthesis. An increase in free acid
and a decrease in total acid content is a positive indication for the antiulcer
activity of the administered compound.
The leaves extract of Sesbania grandiflora L. prevented acute gastric
mucosal injury induced by aspirin. The leaves extract did not modify the
volume, pH and hydrochloric acid content of gastric secretion. It is known that
pepsin requires acidic digestion medium for protein digestion. At pH of
1.6-3.2, the pepsin is most active. Its digestive enzymatic activity decreases
as the gastric pH increases. The lack of change in gastric secretion parameters
observed with Sesbania grandiflora L. leaves extract may be
important because it may not interfere with the process of food digestion and
the absorption of diet proteins, avoiding opportunistic infections of the
digestive tract.
The mechanism by which
this extract produces antiulcer effect is not entirely clear, but the
preventive antiulcer activity of Sesbania grandiflora L. as showed in this study may be ascribed to the presence
of tannins and triterpenes.
Tannins have astringent
action, precipitating proteins of mucosal membranes and skin. Some tannins suppresses the gastric secretion, having a local
action of protection of the gastric mucosa in stress-induced gastric lesion in
rats.
In aspirin experiment,
the action of the extract, when administered, the injury produced by revealed
that probably not only tannins may be responsible for the antiulcer action.
On the other hand, some
triterpenes are known as antiulcer agents and its
action has been mentioned to be due to activation of cellular protection,
reduction of mucosal prostaglandins metabolism, citoprotective
action and reduction of gastric vascular permeability.
The extract also
revealed an important protective effect on gastric mucosa. Doses of 100 mg/kg
of leaves extract administered for prevention of gastric lesions in the test
did not show any pharmacotoxic signs (diarrhea, piloerection, stereotypic activity, convulsions,
depressive, excitatory or sleepiness symptoms) and were not sufficient to
establish the exact value of DL50. This is due to the insufficient amount of
extract available to achieve the very large doses that would be necessary. The
results of the present study seem to provide a support for the use of Sesbania grandiflora L. as an antiulcer drug in folk
medicine.
DISCUSSIONS:
The ethanolic
extract of Sesbania grandiflora L. prevented acute gastric
injury in rats. Stress and nonsteroidal
anti-inflammatory drug induced lesions were significantly prevented by the
extract. At the doses used, the
animals had no depressive, excitatory
or sleepiness symptoms, suggesting that probably centrally acting components
involved in antiulcer action are not found in the extract. The results suggest
that Sesbania grandiflora L. has antiulcer potential.
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Harsh Mohan.
Textbook of Pharmacology. Jaypee Brothers Medical
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Hoogerwerf WA and Pasricha PJ.
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Sura S, Anbu JS and Mahesh U.
Antiulcer Effect of Ethanolic Leaf Extract of Avicennia officinalis. Pharmacologyonline.
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Received on 22.09.2012
Modified on 15.10.2012
Accepted
on 25.10.2012
© A&V Publication all right reserved
Research Journal of Pharmacognosy
and Phytochemistry. 4(6): November –December 2012, 322-325