Pineapple Juice Administration and Gastric Ulcer in Wistar Rats
Fruits have been part of the human diet and supplements. Present study was conducted to investigate pineapple juice (PJ) effects on Gastric Ulcer (GU), since dietary substances or supplements may predispose someone to GU. Thirty-six male wistar rats, weighing 180-200 g were used. They were divided into six groups A-F. Groups A served as control. Group B was not given PJ. Groups C-F received 0.5, 1.0, 1.5 and 2 mL of PJ, respectively. After 30 days, GU was induced with indomethacin. Ulcers Indices (UI) were scored, Total Protein Content (TPC), Catalase (CAT), Superoxide Dismutase (SOD) activities and Lipid Peroxidation (LPP) were determined from stomachs tissue. Significances were taken at p<0.05. UI showed significant increases when groups B-F were compared with control. A significant reduction was seen in TPC when groups B-F were compared with control. Comparison of CAT activities between control and groups B-F showed significant reduction in groups B-F and a significant increase in SOD in groups B-F. Activities of anti-oxidant enzymes in group B were compared with groups C-F. Observation showed that CAT activity increased significantly in groups C and D and SOD activity reduced significantly in groups D-F. Observation from LPP in the gastric mucosa showed increase which was significant in group B, when control was compared with groups B-F. When group B was compared with groups C-F, significant increases were observed in groups C and D. Conclusion from results suggested that PJ consumption may predispose, consumer to gastric ulceration because of its tendency to reduce TPC, increase UI and anti-oxidant enzymes.
Received: April 02, 2013;
Accepted: May 15, 2013;
Published: July 04, 2013
Over the years fruits have been part of the human diet and food supplements.
They are considered as healthy food supplements because they contain high quantity
of water, carbohydrates, proteins, vitamins A, B1, B2,
C, D and E; and minerals such as Ca, Mg, K, Zn and Fe (Okwu
and Emenike, 2006). Fruits are recommended internationally as nutrient supplements
in addition to their dietary importance. Nutritionists have advised that eating
at least five portions of fruits and vegetables daily can help people to maintain
good health throughout their lives, protecting them from heart disease and cancer
(Wenkam, 1990; Food Commission, 2009).
Fruit consumption has been reported to be beneficial to health and contribute
to prevent degenerative processes, particularly lowering the incidence and mortality
rate of cancer and cardio-and cerebro-vascular diseases (Rapisarda
et al., 1999). A diet rich in fruit and vegetables was associated
with smaller gains in body mass index (BMI) (Newby et
Pineapple (Ananas comosus) can also be referred as the King of
Fruit (Bartolome et al., 1995). Hawaii,
Philippines, Caribbean area, Malaysia, Taiwan, Thailand, Australia, Mexico,
Kenya and South Africa have been known to cultivate pineapple extensively. Pineapple
juice was found to be a rich source of vitamin C which is effective in killing
parasites such as worms. It is rich in maganese and is needed for the body to
build bone and connective tissue. Pineapples have been used in traditional tropic
medicine for aliments ranging from constipation to jaundice and to relieve sore
throats and bronchitis. Heart patients have found pineapple useful but coagulation
time has been found to increase which is not good for those with kidney and
liver diseases (Nieper, 1977). In 1891, an enzyme called
bromolian was isolated from the flesh or the pineapple and discovered to comprise
a complex mixture of sulfhydryl containing proteolylic enzymes in addition to
a number of non-specific components like glycoproteins, phosphatase, peroxidases,
glycosidases, celllulases and carbohydrates. The extract also contains a proteinase
inhibitor consisting of inhibitors (Morita et al.,
1979). Pineapple juice also destroys harmful proteins in the stomach and
intestines. It promotes the digestive processes because only protein compound
that have broken down into smaller components can be absorbed by the intestines
and enter the bloodstream (Kelly, 1996). Pineapple
juice has been found to be effective in dissolving mucus and assisting recovery
from tuberculosis (Taussig and Nieper, 1979; Tochi
et al., 2008).
It is known that the human gastrointestinal tract serves two main purposes;
the main doorway for the nutrients and also as a barrier to the external environment.
The duality of this pattern may prevent the great effect of some nutrient to
preserve the integrity and function of the gastrointestinal mucosa itself (Roberfroid,
2000). The epithelial cells of the gastrointestinal tract are the primary
boundary between the ingested nutrients, the blood and the lymph streams. The
cells of the gastrointestinal tract that depend on the luminal and blood stream
flow include, the epithelial cells, mucosal cells, enteroendocrine cells, intraepithelial
lymphocytes and the multiple cells types of the laminal propria. Functional
food describes food or nutrients whose ingestion leads to physiological
changes which are quite separate and distinct from those associated with their
roles as nutrients (Koletzko et al., 1998).
Gastric ulcer is a form of peptic ulcer characterized by the erosion of the
mucosal surface of the stomach. Peptic ulcer is due to the sum of two groups
of forces (defensive and offensive) acting upon the gastrointestinal mucosa.
The most aggressive factor is the hydrogen ions, the acid produced by the parietal
cells in the stomach and pepsin, while the defensive force is from the mucosal
wall and mucus; being the pathogenesis of gastro duodenal ulcer. A large number
of people in the world are affected by gastric and duodenal ulcers. Some of
the causes of the these disorders are: dietary intake (tea, pepper, beverage)
stress, smoking, nutritional deficiencies and ingestion of non-steroidal anti-inflammatory
drugs (Nash et al., 1994; Basil
and Howard, 1995; Ibu et al., 1986; Olaleye
et al., 2006; Ibironke et al., 1997).
One of the predisposing factors of gastric ulcer is dietary intake: similarly
improper eating habits may result into gastro-intestinal dysfunctions (Okike,
1997), uncontrolled eating habit has brought about sickness (Amure,
1990), diet and dietary habits have been shown to be linked to most gastrointestinal
disorders, however beneficiary a diet may be if proper precaution were not taken
(Hyams, 1983). Due to the presence of sorbitol of fructose
in excess of glucose in apple and pear juices, the malabsorption was taken as
a factor in chronic nonspecific diarrhea (Thomas, 1957;
Kelly, 1980). Similarly gastric emptying is highly influenced
by volume of the stomach and volume ingested, calories, osmolality, the amount
of acid that the stomach produced, as well as other factors associated with
physiological factors e.g., splanchnic blood flow, body position, posture and
electrolyte balance (Moore et al., 1984; Fraser
et al., 1990; Marzio et al., 1991;
Horowitz et al., 1993; Amidon
et al., 1991).
Pineapple is a non-citrus fruit with attractive flavor and refreshing sugar-acid
balance (Bartolome et al., 1995). It is available
fresh or canned and as juice (Bartolome et al., 1995).
A 100 g pineapple contains 47-52 calories, 85.3-87.0 g water, 0.4-0.7 g protein,
0.2-0.3 g fat, 11.6-13.7 g carbohydrate, 0.4-0.5 g fiber, 0.3-0.4 g ash, 17-18
mg calcium, 8-12 mg phosphorus, 125-146 mg potassium, 1-2 mg sodium and 0.5
mg iron (Duke, 1983). Pineapple contains 12-15% sugars
(2/3 sucrose% 1/3 glucose and fructose). Pineapple contains between 0.6 and
1.2% of acid (87% citric acid and 13% malic acid) (Samson,
1986; Adhikary et al., 1987). Pineapple
has a pH of 3.71 which is acidic and the acidity percentage is 53.5%. Geographical,
cultural and seasonal harvesting and processing determine the composition of
In view of the composition of pineapple juice vis-α-vis gastric disorder, this study is conducted to investigate the involvement of pineapple fruit juice consumption in the etiology of ulcer because of its wide range of uses and composition.
MATERIALS AND METHODS
Preparation of pineapple fruit juice: The pineapple fruits were obtained from new Benin market in Edo State of Nigeria. The juice was extracted with a juice extractor after peeling. The coring and macerating of the pineapple fruit was done by the juice extractor. The juice was collected and stored in a clean bottle kept inside a refrigerator (deep freezer) for preservation before administration to the rats.
Animals grouping and administration of pineapple: Thirty-six male Wistar
rats weighing between 180-200 g were used for this study. The animals were obtained
from an animal house in Benin, Edo state Nigeria. They were transferred to the
animal house of Igbinedion University Okada. They were kept in separate cages
of six animals per cage constituting a group. They were allowed four weeks to
acclimatize, prior to the commencement of the experiment. The animals in groups
were labeled from A to F. Group A and B were both control groups for this study.
They were given free access to water under standard condition of temperature
and humidity. Group A were animals not given pineapple juice and ulcer not induced.
Group B were animals not given pineapple juice but ulcer was induced in them.
Group C were animals given 0.5 mL of pineapple juice extract and latter induced
ulcer. Group D were animals which were given 1 mL of pineapple juice extract
and latter induced ulcer. Group E were animal which were given 1.5 mL of pineapple
juice and latter induced ulcer. Group F were animal given 2 mL of pineapple
juice and latter induced ulcer. Pineapple juice administration lasted for twenty-one
Ulcer induction and determination: Ulcers were induced with indomethacin
dissolved in Sodium bicarbonate solution of 5 mg per 500 mL distilled water.
Indomethacin was administered at 40 mg kg-1 b.wt., after 24 h that
food had been withdrawn from them. Four hours after, the animals were opened
under anesthesia (Sodium Pentobarbitone) at 60 mg kg-1 b.wt. intra-peritoneally
and pyloric ligation of the stomach was performed. At another 4 h the stomachs
were removed and opened along the greater curvature, washed in normal saline
to remove debris and pinned on a cork for ulcer scoring. The wound in the glandular
were located with a hand lens. Ulcers were scored using the typical criteria
as shown in Table 1 (Rifat-uz-Zaman et
al., 2004; Tanaka et al., 1993).
Tissue preparation: The mucosal tissue from each animal was scraped from the stomach with a blunt knife and the tissue was weighed, transferred to the ice-cooled test tube and homogenized. The homogenate was then centrifuged at 12,000 rpm at 4°C, for 10 min. Supernatant aliquot and stored at-70°C until determination of total protein, Catalase (CAT), Superoxide Dismutase (SOD) and Malondialdehyde (MDA).
Total protein determination: The protein content of the tissue samples
was estimated by the method of Lowry et al. (1951)
using bovine serum albumin as a standard.
Determination of catalase activity: Activity of catalse in gastric mucosa
was determined according to the procedure of Goth (1991)
by following the absorbance of hydrogen peroxide at 230 nm and pH 7.0.
Determination of Superoxide dismutase activity: SOD activity in the
gastric mucosa was determined by measuring the inhibition of autooxidation of
epinephrine at pH 10.2 at 30°C by the method of Misra
and Fridovich (1972). One unit of SOD activity represents the amount of
SOD necessary to cause 50% inhibition of adrenaline autooxidation.
|| Criteria for scoring ulcer
|| Ulcer index, total protein content, anti-oxidant activities
and lipid peroxidation upon administration of pineapple juice in wistar
|*Shows significance at p<0.05 when groups were compared
with animals in group A, Shows significance at p<0.05 when groups
were compared with animals in group B
Malondiaidehyde determination: Lipid peroxidation of gastric mucosa
was determined spectrophotometrically at 533 nm and MDA concentration was quantified
by using the molar extinction coefficient, 1.56x105 moL-1 E cm-1
(Buege and Aust, 1978; Nair et
Statistical analysis: All values presented in tables are expressed as Mean±SEM. The appropriate comparisons between groups were made using studentst-test. The differences between the groups were taken to be significant at p<0.05.
From the study, groups C, D, E and F were compared with groups A and B independently.
Table 1 showed the criteria that were used to score ulcer
in this study as used by Rifat-uz-Zaman et al. (2004)
and Tanaka et al. (1993). From the experimental
results showed in Fig. 1, Ulcer indices scored in the various
rats groups were statistically compared. The results showed that ulcer inductions
in the various groups of rats were significantly increased in group C (15.75±0.50),
D (24.92±0.82), E (12.17±0.59) and F (11.08±0.83) when
the groups of rats, not given pineapple juice (groups A (0.00) and B (10.25±0.50))
were compared with the groups of rats that were given pineapple juice. But the
highest value of significance was noticed in group D rats where 1 mL of pineapple
juice was administered.
This is in agreement with the observed trends in ulcer model with respect to Total protein contents in groups B (1.40±0.78), C (1.65±0.13), D (1.41±0.61), E (1.41±0.11) and F (1.62±0.05) when compared with group A (2.38±0.18). The results showed reductions that are not statistically significant. Similarly animals in groups C, D, E and F were compared with group B animals, the result showed a slight increase not higher than group A. But in all, no statistical significance was recorded. Other biomarkers used to justify the recorded ulcer indices were catalase, superoxide dismutase and Malondialdehyde. They were all shown in Table 2.
||Effect of pineapple consumption juice on ulcer index in rats
Catalase activities in the groups of the rats, showed a general reduction when group B (2.45±0.49), C (5.40±0.67), D (3.21±0.24), E (4.08±0.51) and F (3.69±0.52) were compared with group A (5.49±0.14). Significant reductions were recorded in group B, D, E and F. Comparison of groups C, D, E and F with group B showed the activity of Catalase to increase with significant increase in group C and E.
Results from superoxide dismutase activity in the rats were compared. Groups B (5.80±0.11), C (6.37±0.84), D (4.31±0.25), E (5.00±0.50) and F (4.34±0.59) were compared with group A (2.80±0.13). The observed result showed significant increases. In the other way, group C, D, E and F were compared with group B. Recorded observations were reductions in activities with groups D, E and F with significance in groups D and F. The increase observed in group C was not significant.
Comparisons of the lipid peroxidation of the gastric mucosa in the rats were
also shown in Table 2. Groups B (3.03±0.36), C (5.10±0.21),
D (4.48±0.46), E (3.50±0.20) and F (3.33±0.47) were compared
with group A (2.15±0.28). General increases were recorded with a significant
increase in group B. Similarly group B was compared with groups C, D, E and
F while the recorded observation were increases in all the groups (groups C,
D, E and F), which were only significant in groups C and D.
Despite the fact that fruits contained useful food components, minerals and
vitamins (Okwu and Emenike, 2006), the results from this
study clearly showed that pineapple juice has a tendency to predispose one to
gastric ulcer. Analysis of the ulcer indices revealed that the various volumes
of pineapple juice promoted gastric ulceration because of the significant increases
indicated when they were compared as shown in Fig. 1. This
is consistent with other results of earlier researchers in this field who had
implicated some useful dietary food substances in the etiology of gastric ulcer
(Nash et al., 1994; Basil
and Howard, 1995; Ibu et al., 1986; Olaleye
et al., 2006, Ibironke et al., 1997).
A reduction in total protein content, confirmed further that despite the wide
medicinal usefulness of pineapple juice, it does not seem to have sufficient
capability to protect the gastric epithelia layer from being eroded away. Pineapple
juice has been reported to be effective in dissolving mucus and aiding recovery
from the dread disease in tuberculosis patient (Taussig
and Nieper, 1979; Anandan et al., 1999;
Olaleye et al., 2006), similarly the juice has
been reported to be acidic (Samson, 1986; Adhikary
et al., 1987). In view of this, we can propose that pineapple juice
may also have some interaction with the mucus content of the stomach dissolving
the mucus thereby exposing the stomach to attack.
Comparison of catalase activities in group A with other groups B, C, D, E and F showed that pineapple juice can promote gastric ulceration as indicated by a reduction in activity. When control animals in groups B (2.45±0.49) were compared with the animals in groups C, D, E and F, increase in activity was noticed, but this increase did not exceed the value obtained for the control group A (5.49±0.67) and became significant in groups C (5.40±0.14) and E (3.69±0.52). These combined effects suggested that pineapple juice protective role under this condition may not be sufficient enough to prevent gastric ulceration.
When superoxide dismutase activities were compared as shown in Table 2, it revealed increased activities that were significant when group A animals were compared with animals in groups B, C, D, E and F. But the comparison of groups C, D, E and F with group B brought similar result which is consistent in a typical gastric ulcer model with antioxidant enzyme (superoxide dismutase). There was reduction in activity in all the groups except group C in which 0.5 mL of pineapple juice was administered.
Observation recorded that lipid peroxidation with malondialdehyde showed increases
when groups A and B were compared with other groups (i.e., group A compared
with B, C, D, E and F; group B compared with groups C, D, E and F).These observations
clearly showed that gastric mucosal damage may be by hemorrhagic lesions through
oxidative damage of the mucosa by the increased lipid peroxidation, decreased
level of antioxidant defensive enzymes and decrease in protein content (Bandyopadhyay
et al., 1999; Priya et al., 2012).
Lipid peroxidation leads to loss of membrane fluidity, ion transport and membrane
integrity of the surface epithelia cells which lead to gastric lesions. The
lipid content also determines the degree of resistance of mucin to peptic degradation
and thus contributes significantly to mucus viscosity hydrophobicity and impedance
to hydrogen ion diffusion (Bilski et al., 1987;
Olaleye and Farombi, 2006).
Characteristics model of gastric ulceration has shown reduction in total protein,
increase in anti-oxidant activities and reduction in lipid per-oxidation. But
results from this experimental study suggested that Pineapple juice consumption
may predispose the user to gastric ulceration because it has the tendency to
reduce total protein content and increases anti-oxidant enzymes activities.
Although, effectiveness of the lipid per-oxidation can not be ascertained because
two out of the three conditions studied clearly showed that Pineapple juice
consumption can promote gastric ulcer and similarly pineapple juice was found
to be effective in dissolving mucus (Taussig and Nieper,
1979; Anandan et al., 1999). Therefore,
one should consume the juice with caution.
In conclusion, the results of this study clearly showed that despite many medicinal values and nutritional benefits that one can derive from eating pineapple and its juices, care must be taken because it can predispose one to gastric ulceration. However studies are going on to investigate other interactions that pineapple juice may have with our body.
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