Journal of Rawalpindi Medical College (JRMC); 2014;18(2):286-289
Original Article
Protective Effect of Vitamin E on Indomethacin Induced
Gastric Ulcers
Ayesha Yousaf,*Shaista Ali,*Zahra Haider Bokhari,** Attiya Mubarak Khalid,*** Muhammad Amin Sheikh.****
*Department of Anatomy, Rawalpindi Medical College, Rawalpindi. ** Department of Anatomy, King Edward Medical
College, Lahore. *** Department of Anatomy, Avesina Medical College, Lahore. **** Department of Anatomy, SIMS, Lahore.
Abstract
by surface epithelial cells and mucus neck cells of
gastric glands.2
Indomethacin is anti-inflammatory drug that
inhibits the synthesis of prostaglandins. 3,4 Inhibition
of prostaglandin synthesis leads to gastrotoxic effect
of indomethacin.5 Prostaglandins stimulate mucus and
bicarbonate secretion, inhibit gastric acid secretion to
maintain pH gradient.6 Gastric mucosal protection is
impaired by decreased prostaglandin level and
increases
secretion
of
acid
to
produce
ulcer.7Indomethacin produces lesions in the pylorus as
it is the most vulnerable part. The pyloric antral lesion
may reach maximum size in 6-10 hours, penetrated
muscularis mucosae within three days and remains
for at least 7 days.3 Vitamin E is a lipid soluble
antioxidant
that
increases
secretion
of
prostaglandins.8,9 Vitamin E activates enzyme
phospholipase A2 and increases the synthesis and
secretion of prostaglandins.10
Background: To study the histomorphological
changes in gastric mucosa induced in mice by
Indomethacin and to assess the protective effect of
vitamin E with variable duration of drugs.
Methods: In this experimental study 48 adult
albino mice were divided randomly in to four groups
A, B, C and D each group comprising of 12 mice.
Group A was the control group. Group B was given
Vitamin E 400mg/kg. Group C was given
Indomethacin 25mg/kg and Group D was given a
combination of Indomethacin and Vitamin E
(25mg/kg & 400 mg/kg respectively). In each group 6
animals were given dose for 3 days and other 6
animals for 8 days. They were sacrificed 24 hours
after the last dose. After dissection gastric mucosal
changes regarding colour and site of ulcers were
examined under dissecting microscope. Histological
parameter observed was presence/absence of mucus
in the cells of gastric glands.
Results: Findings were normal in groups A and B.
Reddish mucosa with ulcers in pyloric part of
stomach was seen in groups C and D. Decreased
mucus in cells of gastric glands was also observed in
group C as compared to group D.
Conclusion: Severity of gastric ulcers induced by
Indomethacin in mice is decreased by giving
Vitamin E which protects gastric mucosa by
increasing mucus in cells of gastric glands.
Key Words: Indomethacin, vitamin E, gastric
mucosa, Non-Steroidal Anti-inflammatory Drugs
Materials and Methods
The study was conducted in the Anatomy
Department of King Edward Medical University and
Post Graduate Medical Institute, Lahore. Colonies of
Mus-musculus Swiss Webster albino mice were
procured from Veterinary Research Institute of
Lahore.They were acclimatized for 14 days in Post
Graduate Medical Institute, Lahore. 48 adult healthy
male mice of body weight ranging from 20 to 35gm
used in the study were divided at random into a
control group-A and experimental groups-B, C and D
each comprising of I2 animals. Groups were further
subdivided into subgroups A1, A2, B1, B2, C1, C2, D1
and D2 comprising of 6 mice each according to
duration of drug. Groups of mice were kept in cages
which were labelled as A, B, C and D.
Indomethacin was given in a dose of 25mg/kg body
weight/day. Stock suspension of Indomethacin was
prepared daily by mixing finely ground powder of
one capsule in 2ml of distilled water by constant
stirring. Vitamin E was given in a dose of 400mg/kg
body weight/day. Stock suspension was prepared
Introduction
Mice as well as human stomach have a similar
histology. Stomach wall is composed of mucosa,
submucosa, muscularis externa and serosa. Mucosa is
composed of epithelium, lamina propria and
muscularis mucosae. Gastric mucosal defense includes
mucus-bicarbonate-phospholipid barrier and surface
epithelial cells generating bicarbonate, mucus,
phospholipid and prostaglandins.1 Mucus is produced
286
Journal of Rawalpindi Medical College (JRMC); 2014;18(2):286-289
daily by mixing liquid of one capsule in 2ml of corn
oil. The indomethacin and Vitamin E suspension was
given orally according to their body weight by feeding
tube. The daily dose of Indocid and Evion for each
mouse was calculated by formula: Dose (in ml) = 0.002
x Wt (Wt = weight of the animal in gm)
Group A was the control group. Group B was
given Vitamni E 400mg/kg. Group C was given
Indomethacin 25mg/kg and Group D was given a
combination of both (25mg/kg & 400 mg/kg
respectively). A1, B1, C1 and D1 were given dose for 3
days and A2, B2, C2 and D2 for 8 days. Animals were
sacrificed next day after the last dose. Stomach was
incised along the lesser curvature and the interior was
washed with normal saline. It was then examined
under dissecting microscope for gross parameters. The
colour of mucosa of fundus, body and pyloric part of
stomach in experimental groups was compared with
the control group. Normal colour of gastric mucosa in
group A and B was from greyish pink to pink. Colours
varying from greyish pink to reddish pink were
observed. Stomach of animals of each subgroup was
put into separate labeled glass bottles containing 10%
formalin fixative fluid for 48 hours. Each stomach was
cut transversly into two halves, separating body from
the pylorus. Strips measuring 2×4mm having ulcers
were cut from the pyloric part of the stomach. After
routine processing and staining presence/absence of
mucus was seen in the cells of gastric glands by using
PAS staining at 10X and 40X magnifications.
B1 and B2 (Table.3, Fig. 2).In subgroups C1 and C2 the
mucus was totally absent from the glands (Table.3,
Fig. 2). Mucus was seen in 5 specimens of D1 and D2
each whereas it was absent in 1 specimen of D1 and
D2 (Table.3, Fig. 2).
Table 1: Color of gastric mucosa in study groups
Color of gastric mucosa
Dull
Reddish
Pinkish
red
pink
0
0
0
0
0
0
1
0
0
5 (83.3%)
(16.7%)
2 (33.3%) 4 66.7%)
0
0
0
0
3 50%)
3 (50%)
3(50%)
0
0
3 (50%)
2 (33.3%) 2(33.3%)
0
2 (33.3%)
2 (33.3%) 2 33.3%)
0
2 (33.3%)
Study
Groups
A1
A2
B1
Greyish
Pink
6 (100%)
6 (100%)
B2
C1
C2
D1
D2
Total
23(47.9%)
9 (18.9)
6(12.5%)
10(20.8%)
Table 2: Site of ulcers in study groups
Site of ulcers
Pyloric
Study groups
Results
In control group A1 and A2 color of gastric mucosa
was normal i-e greyish pink. Mucosa appeared shiny
and transparent. No vascular congestion was seen.
The mucosa in 5 subjects of B1 and 2 subjects of B2
were greyish pink while pink color was seen in 1
subject of B1 and 4 subjects of B2. The mucosa was
shiny and transparent and no sign of hyperemia was
noted (Table 1, Fig. 1).The gastric mucosa was dull red
in 3 subjects each of C1 and C2. Reddish pink mucosa
was noted in 3 subjects each of C1and C2. They lacked
luster. Mucosal surface was congested (Table 1, Fig.
1).In 2 subjects each of D1 and D2, the gastric mucosa
was greyish pink. Pink color was observed in 2
subjects each of D1 and D2. The mucosa was shiny
and transparent and no hyperemia was seen. It was
reddish pink in 2 subjects of D1 and D2 and was
lusterless and a little congested (Table 1, Fig. 1).
Histological parameters revealed presence of mucus
in cells of glands of all slides of subgroup A1 and A2
(Table.3, Fig. 2).Mucus was seen in glandular cells of
A1
0
A2
B1
B2
C1
C2
D1
D2
Total
0
0
0
6 (100%)
6 (100%)
2 (33.3%)
2 (33.3%)
16 (33.3%)
p-value=0.000- Significant
Table 3:Mucus in gastric glands in study
groups( p-value=0.000-significant)
Study groups
A1
A2
B1
B2
C1
C2
D1
D2
Total
287
Mucus in the gastric glands
Present
6 (100%)
6 (100%)
6 (100%)
6 (100%)
0
0
5 (83.3%)
5 (83.3%)
34(70.8%)
Absent
0
0
0
0
6 (100%)
6 (100%)
1 (16.7%)
1(16.7%)
14 (29.2%)
Journal of Rawalpindi Medical College (JRMC); 2014;18(2):286-289
strengthens the gastric mucosal barrier in animals
through its antioxidant effect.10, 11 In the present study,
the protective effects of vitamin E were evaluated
against experimentally induced gastric mucosal
lesions in mice. Color of gastric mucosa changed from
greyish pink to reddish pink in group given
Indomethacin. This corroborated with the findings of
earlier studies who observed that high concentration
of Indomethacin in the blood resulted in uncoupling
of endothelial cell DNA. Cell death caused vascular
damage, thrombosis and the tissue ischemia leading to
necrosis.5,12
All ulcers were located in pyloric part. No ulcer was
seen in body of stomach. This finding in the current
project is statistically significant (p-value < 0.05) and
corroborated with findings of Elahi et al.13 Gastrin is a
peptide hormone produced by G cells in pyloric
antrum that stimulates parietal cells to secrete HCl/
gastric acid.14 It is speculated that increased level of
HCl in pyloric part with superadded Indomethacin
induced increased gastric secretions make pyloric part
more susceptible.
A significant reduction/absence of mucus content
in gastric glands was noted in group given
Indomethacin as compared to group given Vitamin E
along with Indomethacin. This is in accordance with
the findings that Indomethacin decreased secretion
and thickness of the mucus layer.2 These findings
suggest that when gastric surface mucus cells are
exposed to acid, gastric epithelial permeability
decreases to inhibit acid back diffusion.15 This is in
accordance with researchers who stated that Vitamin
E increases the amount of mucus by stimulating
prostaglandins synthesis and secretion.10,16
Fig.1. A Photograph of pylorus of stomach observed under
dissecting microscope depicting color of mucosa in study
groups;Group A. Greyish pink mucosa;Group B. Pinkish
mucosa;Group C. Dull red mucosa;Group D. Greyish pink
mucosa;All ulcers of C and D subgroups were seen in
pyloric part of stomach
.
Conclusion
Vitamin E has a protective effect on gastric mucosal
injury by increasing mucus secretion and by
increasing secretion of prostaglandins.
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Fig 2. A Photomicrograph of stomach pylorus showing
mucus in cells of gastric glands (PAS stain,
Magnification 900X);Group A. Normal mucus in gastric
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Increased mucus in gastric
glands;Group C. No mucus in gastric glands;Group D.
Increased mucus in gastric glands
2.
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4.
Discussion
The deleterious effects of Indomethacin on different
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