Carica Papaya as a Mucosa Enhancer in Gastric
Lesion after Ethanol Administration in Adult
Male Wistar Rats
Okesina AA1*, Mohammed AA2, Buhari MO and Caxton Martins EA3
1Department of Anatomy, University of Gitwe, Nigeria
2Department of Human Anatomy Kampala International University Western Campus Ishaka Bushenyi, Uganda
3Department of Anatomy, University of Ilorin, Nigeria
Submission: September 12, 2017; Published: November 27, 2017
*Corresponding author: Okesina Akeem Ayodeji, Department of Anatomy, University of Gitwe, Rwanda, Nigeria, Tel: 08033725777;
Email:
Introduction
Carica papaya (CP) is a member of the family Caricaceae, the
third most important horticultural crop after citrus and plantain
in Nigeria [1-5]. CP is a dual or multi-purpose, early bearing, space
conserving, herbaceous crop, is widely acclaimed, despite its
susceptibility to natural enemies [4]. In some islands of the West
Indies, it is known as pawpaw [1]. Originally from Southern
Mexico, Central America and Northern South America, the papaya is
now cultivated in most countries with tropical climate like Malaysia
and the West Indies. Caricapapaya leaf (CPL) is used for food or as
medication in folk medicine. It is consumed as a vegetable by the
Malay community in Malaysia and by the natives of the East Indies.
Traditionally, the leaf extract was used as a tonic for the heart, an
analgesic and in the treatment of stomach ache [2]. The extract is
also known to have antioxidant properties [6], but there are no
induced acute gastric damage. Gastric ulcers, one of the most
widespread disease states, are believed to be due to an imbalance
between acid and pepsin along with weakness of the mucosal
barrier. There are many products used for the treatment of gastric
ulcers, such as antacids, proton pump inhibitors or antihistaminic
agents, but most of these drugs produce several adverse reactions.
Thus, there is a need for more effective and less toxic anti-ulcer.
Materials and Method
Animals
24 adult male Wistar rats weighing 190-210g, which were
purchased from Animal house University of Ilorin were used in this
project work. They were each weighed and randomly assigned to
three groups (A, B and C). They were kept and maintained under
standard laboratory conditions of temperature, humidity, and light.
They were allowed free access to standard laboratory diet pellets
and water liberally. All animals were fasted 24 hours before the
commencement of the experiment. Ethical approval was obtained
from the University of Ilorin ethical review committee.
Research Article
Volume 8 Issue 2 - November 2017
DOI: 10.19080/ARGH.2017.08.555732
Adv Res Gastroentero Hepatol
Copyright © All rights are reserved by Okesina Akeem Ayodeji
Adv Res Gastroentero Hepatol 8(2): ARGH.MS.ID.555732 (2017) 001
Abstract
This study was performed to determine the regenerative potential of Cariaca papaya leaves (CPL) on 50% ethanol induced gastric lesions.
It involves histological studies, including Haemotoxylin and Eosin, Periodic Acid Schiff (PAS), Aldehyde Fuchsin, and Van Geison’s stain, in the
stomach of male adult Wistar rats. Twenty-four male adult Wistar rats weighing 180- 220g were used in this study, with the animals divided
into three groups (n=8 per group). The control group (group A; n=8) received phosphate buffered saline orally, with the aid of a cannula. 1 ml of
50% ethanol was administered orally, by a cannula, to produce the gastric lesion in group B (n=8). And in the group C, 500mg/kg body weight
of paw paw leaf extract was administered orally, with a cannula, twelve hours after ethanol administration to the rats. This experiment lasted
for twenty one consecutive days.
Histological studies showed the rat’s gastric mucosa to be eroded, following ethanol administration; and after the treatment with CPL, some
form of regeneration was observed.
Keywords: Caricapapaya; Ethanol; Mucosa; Hematoxylin
Advanced Research in Gastroenterology & Hepatology
How to cite this article: Okesina A, Mohammed A, Buhari M, Caxton M E. Carica Papaya as a Mucosa Enhancer in Gastric Lesion after Ethanol
Administration in Adult Male Wistar Rats. Adv Res Gastroentero Hepatol 2017; 8(2): 555732. DOI: 10.19080/ARGH.2017.08.555732.
002
Plant Material and Extract Preparation
Leaves of Carica papaya leaves were collected within the
premises of College of Medicine University of Ilorin Kwara State
of Botany University of Ilorin. A large quantity of leaves were
collected, washed clean with water, air dried and reduced to powder
using a mortar and pestle. Powdered leaves of plant weighing 100g
was mixed with 1litre of 70% ethanol and the mixture was left for
a water bath (maintained at 60±0.2 °C) to yield 15g of dark green
solid extract. The solid extract was stored in a refrigerator.
Treatment
Group A received oral phosphate buffered saline (PBS) and
served as control. Group B (Ethanol) received 1.0ml of 50% ethanol
orally every 12 hours. Group C (Ethanol+CPL) received pawpaw leaf
extract (500mg/kg) orally 12 hours after 1.0 ml 50% ethanol for 21
consecutive days. 24 hours after the last administration the animals
sections at 5µm thickness was obtained on a Leitz rotary microtome
and stained with Haematoxylin and Eosin (H & E), Van geison’s stain
for elastic tissues, Aldehyde Fuchsin, Periodic Acid Schiff for brush
borders.
Results
Physical observation
During the time of this investigation the animals were carefully
observed as regards to feeding and physical appearance. Feeding
in the animals was not altered as they were still feeding normally.
Feeds were weighed before it was given to the animals and the
quantity left in the feeding though, if any, was measured. The eyes
remained pinkish and also the faces were observed to be normal
that is dark brown and solid but not too dry.
Weight of the animals
Figure 1: The average percentage weight change in the 24
experimental animal (n= 8) at the end of the experiment.
The animals were weighed with an electronic weighing balance
(SALTUN@EK5055Max) during and after the experiment, and
the average weight of the three groups (Group A which is the PBS
control group, group B which is the Ethanol treated group and
group C which is the Ethanol+CPL treated group) in percentage
were compared at the end of the experiment. After 21 days of
administration, the weight of the rats were noted and measured
in percentage weight change. The weight of the PBS control group
increased by 0.38%, that of ethanol group was reduced by 4.83%
and the CPL treated group was reduced by 2.33% (Figure 1).
Histological Observations
Figure 2: Photomicrograph of rat stomach (PBS control),
Aldehyde fuchsin stain.M (mucosa) MM(muscularismucosa),
EF(elastic bres), S(submucosa). X320.
Figure 3: Photomicrograph of rat stomach (ethanol treated),
Aldehyde fuchsin stain.M(mucosa) MM(muscularis mucosa),
EF(elastic bres), S(submucosa).X320.
Figure 4: Photomicrograph of rat stomach (CPL treated),
Aldehyde Fuchsin Stain.M(mucosa) MM(muscularis mucosa),
EF(elastic bres), S(submucosa). X320.
Regenerative effect was observed on 1ml of 50% ethanol induced
gastric lesion in rats after the treatment with 500 mg/kg of CPL. The
CPL treated groups showed remarkable changes as compared to
the ethanol treated animals. And these changes produced by CPL
treated groups are comparable to PBS control groups. These results
showed that 500mg/kg of CPL had a regenerative effect against the
gastric lesion caused by ethanol (Figure 2-13).
Advanced Research in Gastroenterology & Hepatology
How to cite this article: Okesina A, Mohammed A, Buhari M, Caxton M E. Carica Papaya as a Mucosa Enhancer in Gastric Lesion after Ethanol
Administration in Adult Male Wistar Rats. Adv Res Gastroentero Hepatol 2017; 8(2): 555732. DOI: 10.19080/ARGH.2017.08.555732.
003
Figure 5: Photomicrograph of rat stomach,(PBS control)
Van Geison’s Stain M(mucosa) MM(muscularis mucosa)
S(submucosa) CF (collagen bers). X320.
Figure 6: Photomicrograph of rat stomach,(ethanol treated)
Van Geison’s Stain M(mucosa) MM(muscularis mucosa)
S(submucosa) CF (collagen bers). X320.
Figure 7: Photomicrograph of rat stomach,(CPL treated)
Van Geison’s Stain M(mucosa) MM(muscularis mucosa)
S(submucosa) CF (collagen bers). X320.
Figure 8: Photomicrograph of rat stomach, (PBS control) H &
E Stain M (mucosa) MM(muscularis mucosa) S(submucosa).
X320.
Figure 9: Photomicrograph of rat stomach,(ethanol treated)H
& E Stain M(mucosa) MM(muscularis mucosa) S(submucosa).
X320.
Figure 10: Photomicrograph of rat stomach,(PBS treated)H &
E Stain M(mucosa) MM(muscularis mucosa) S(submucosa).
X320.
Figure 11: Photomicrograph of rat stomach,(PBS control) PAS
Stain M(mucosa) MM(muscularis mucosa) S(submucosa) GG
(gastric glands). X320.
Figure 12: Photomicrograph of rat stomach, (ethanol treated)
PAS Stain M(mucosa) MM(muscularis mucosa) S(submucosa)
GG (gastric glands). X320.
Advanced Research in Gastroenterology & Hepatology
How to cite this article: Okesina A, Mohammed A, Buhari M, Caxton M E. Carica Papaya as a Mucosa Enhancer in Gastric Lesion after Ethanol
Administration in Adult Male Wistar Rats. Adv Res Gastroentero Hepatol 2017; 8(2): 555732. DOI: 10.19080/ARGH.2017.08.555732.
004
Figure 13: Photomicrograph of rat stomach,(ethanol treated)
PAS Stain M(mucosa) MM(muscularis mucosa) S(submucosa)
GG(gastric glands). X320.
Discussion
In this study the regenerative potential of CPL was investigated
in ethanol induced gastric lesion on adult male Wistar rats. It
has been demonstrated that a single dose (500mg/kg) of CPL
aqueous extract is able to protect the rat gastric mucosa against
haemorrhagic lesions produced by alcohol [3]. This dose was
chosen after preliminary assessment using a wide dose range of
this extract. Oral administration of ethanol in rat is noxious for
the stomach, affecting the gastric mucosa by disrupting its barrier
and provoking pronounced micro/macrovascular changes a few
minutes after its administration
During the experimental period, it was noted that after each
ethanol administration the physical activity of the rats increased
immediately, and after some time they became slow in action. Both
the ethanol and CPL treated groups did not consume their feeds as
much as the PBS treated control group and this must be responsible
for the loss in weight of both groups.
After 21 days of administration, the weight of the rats was
noted and measured in percentage weight change. The weight of
the PBS control group increased by 0.38%, that of ethanol group
was reduced by 4.83% and the CPL treated group was reduced by
2.33%.
Histological observations made after viewing the tissue slides
under the (Olympus) light microscope, revealed three layers of the
rat stomach in this work. Four different stains were used to study
the various structures of the gastric tissues of three different rat
stomachs. The three rat stomachs were divided into PBS control,
Ethanol treated and CPL treated. And four stains were used in this
histological study; Aldehyde fuchsin, Van Geison, H&E and PAS.
present in the mucosa layer. It was densely stained in PBS control,
faintly stained in Ethanol treated and also in CPL treated it was
slight densely stained.
around the muscularis mucosa which stains as red. The red stain
was in form of as a thick line in PBS control while in ethanol treated,
it was a thin line stain. It was also stained thick in CPL treated. H
& E stain was used to investigate the gastric mucosa layer. In PBS
control it was densely stained but not densely stained in ethanol
treated, showing the level of erosion in the mucosa and in the
CPL treated it was densely stained showing level of regeneration.
PAS used to Show the brush boarders, (the mucosubstances) i.e
the gastric glands that produce mucous secreting cells which are
carbohydrate in nature.
PAS positive magenta colour was seen in PBS control, but was
not seen in ethanol treated because it was faintly stained. The
magenta colour was also seen in CPL treated. When comparing
the three groups, regenerative effect was observed on 1ml of 50%
ethanol induced gastric lesion in rats after CPL treatment, which
showed remarkable changes as compared to the ethanol treated
animals. And these changes produced by CPL treated groups are
comparable to PBS control groups. These changes showed that 500
mg/kg of CPL had a regenerative effect against the gastric lesion
caused by ethanol. In summary, the results demonstrated that CPL
treatment exerts regenerative effects in ethanol-induced gastric
lesion in rats.
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Advanced Research in Gastroenterology & Hepatology
How to cite this article: Okesina A, Mohammed A, Buhari M, Caxton M E. Carica Papaya as a Mucosa Enhancer in Gastric Lesion after Ethanol
Administration in Adult Male Wistar Rats. Adv Res Gastroentero Hepatol 2017; 8(2): 555732. DOI: 10.19080/ARGH.2017.08.555732.
005
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