1. 1
CHAPTER ONE
1.0 INTRODUCTON
1.1 BACKGROUND OF STUDY
Gastric ulceration is known as a lesion or scar that occurs on the mucosal epithelium exposing it
to excess gastric acid and aggressive pepsin activity (Khazaeri, 2006).
Ulcer is a well-known problem or disorder accounting for up to 15 mortality out of every
15,000 complications yearly in the world (Shristi, (2012). Even with the advancing management
of the disorder which includes surgical operations such as vagotomy, antrectomy, pyloroplasty,
tying off an artery etc., and normal management techniques which include prostaglandin analogs,
H2 receptor antagonist, proton pump inhibitors and so on.
Moringa oleifera popularly known as Moringa, from the family of Moringacease. It is a fast
growing drought resistance tree, native to the Southern foothills of the Himalayas in
Northwestern India.
But the Moringa made from naturally dried leave is what is going to be used in the course of this
experiment. It has a delicious spinach green flavor and contains a lot of nutrients making it have
a variety of benefits including being a major source of anti-oxidant.
Since ancient times, the medical properties of plants have been investigated in the recent
scientific development throughout the world due to their potent anti-oxidant activities. And as
mentioned earlier Moringa has an abundance of antioxidant, which is one of the many reasons it
is used in this study.

2. 2
Antioxidants have been reported to prevent oxidative damage caused by free radicals, chelating
catalyze metals and also by acting as oxygen scavenger (Shaidi, 1992.). The potentially reactive
derivatives of oxygen species, attributed as reactive oxygen species (ROS), are continuously
generated inside the body. However over production of ROS and inadequate antioxidant defense
can easily affect and persuade oxidative damage to various biomolecules including proteins,
lipids, lipoprotein and DNA (Forber, 1994). This oxidative damage is what can cause gastric
ulceration.
But there are various mechanisms that occur for gastric acid to be secreted.
On the cellular level there is an ATPase pump that initiates/starts and regulates the whole
process. This pump is called the sodium potassium ATPase pump. What this pump does
basically is to provide resting membrane potential for the neurons, because without resting
membrane potential, action potential cannot occur. And action potential is needed for activation
of the nerve cells found in the gastrointestinal tract.
Once the nerve cells are activated at action potential, they go ahead to stimulate the release of
Secretagogues, this secretagogues acts on the stomach or stimulate another pump called the
Hydrogen potassium pump or simply proton pump to release Hydrochloric acid (HCH)
Sodium potassium pump was first observed by Jen Christian Skou in the year 1957 while
working as an assistant professor of Physiology, University of Aarhus, Denmark.
Indomethacin on the other hand is a non-steroidal anti-inflammatory drug (NSAID) that is going
to be used to induce the ulcer in the male Wistar rats.

3. 3
1.2 AIM OF STUDY: The study is aimed at investigating the effect of Moringa and
indomethacin on gastric ulceration, and the gastric parameters to test the effectiveness of the
Moringa on the Male Wistar Rats
1.3 OBJECTIVES: The specific objectives of this study are:
 To investigate the influence of Moringa and indomethacin administration on gastric
ulceration
 To determine the effect Moringa Oleifera would have on the ulcer score, pH of the gastric
juice and oxidative stress level.
 The role that sodium potassium ATPase pump and oxidative stress play in gastric
ulceration.
1.4 STATEMENT OF PROBLEM: Gastric ulcer is known as a lesion or scar that occurs on
the mucosal epithelium exposing it to excess gastric acid and aggressive pepsin activity.
Ulcer is a known problem or disorder accounting for up to/estimate of 15 mortality out of every
15,000 complications yearly in the world. Even with the advancing management of this disorder,
from conventional vagotomy, prostaglandin analogs, H2 receptor antagonist, proton pump
inhibitors, and so on.
But this experiments is going to test the effectiveness of Moringa on ulcer, so as to create a wider
range of solutions to this gastric ulceration.
1.5 JUSTIFICATION: The importance of this study/experiment is to provide more
information on indomethacin, Moringa and its subsequent effect on gastric ulcer and also to
show how oxidative stress and sodium potassium pump are involve in gastric ulceration

4. 4
1.6 EXPECTED CONTRIBUTION TO KNOWLEDGE: This study/experiment is meant to
contribute to the knowledge of gastric ulceration through the tests carried out, investigation and
analysis that would be gotten from the test, providing more information about Moringa and
become the baseline for future studies or research.

5. 5
CHAPTER TWO
2.0 LITERATURE REVIEW
2.1 Gastric ulceration
It is a lesion located basically at the level of the stomach or are known as open sores lining the
stomach. It is a type of peptic ulcer.
It suggest there is an involvement of hydrochloric acid and pepsin in the development of the
disorder. When gastric acid is produced in excess, the mucosal membrane that protects the
stomach from danger are damaged, enabling Helicobacter pylori to penetrate the barrier and
cause internal infection or it will expose the mucosal epithelial membrane to more action of
hydrochloric acid resulting in sores on the membrane from where the acid has come in contact
with it.
So gastric ulceration is the imbalance between aggressive factors (principally secreted gastric
acid and pepsin) and factors that comprise of mucosal defense or mucosal resistance to
ulceration. Gastric ulcer may occur when the aggressive effect of acid and pepsin overweight the
protective effect of gastric mucosal resistance.
It is now established that the generation of free radicals also play an important role in the
formation of gastric ulcer.
And learning the various factors causing gastric ulcer and the mechanism regulating gastric acid
secretion would also help in the understanding and generation of treatment and prevention of the
disorder (gastric ulceration)

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2.2 Factors associatedwith Gastric Ulcer Disease.
This are evidence that shows that this potential risk factors are associated with gastric ulceration.
 Smoking: It is shown by Epidermic studies that cigarrete smokers have an increased
incidence of gastric ulcer disease. It not only increases the chances of getting the disease
but it also slows down or prevents the healing process of the disease. (Hunt, 1980).
Nicotine is also known to reduce pancreatic secretion as well as pyloric sphincter
pressure, and also decreases mucus production. There is a strong association made
between H.pylori infection and cigarrete smoking in patients with and without the gastric
ulcers disease. (Doll, 1958)
 Alcohol: Peptic cirrhosis is associated with an increased risk of duodenal ulcer, but there
is no direct link that alcohol causes gastric ulceration.
But of recent it is shown that dilute (5%) of alcohol has been shown to be a stimulant of
acid secretion. In high concentration (20-40%) of it damages the mucosal membrane,
causing erosion and gastritis. Absolute or total ethanol penetrates the gastric mucosa and
causes cell and plasma membrane permeability.
Furthermore lesions caused by ethanol have contributed you free radical damage, which
results in lipid peroxidation products. (Agarwal, 1980)
 Stress: Stress ulceration of the stomach is now regarded as multifactorial phenomenon.
Acute Stress and Chronic Stress
 Diet: In India duodenal ulcer is more common with rice eating belt than in wheat eating
belt area. (Szabo, 1975). Milk and bland meals are traditional therapies for duodenal
ulcer. But milk is also known to cause ulcer. Coffee, Cola, beverages, beer and wine are
also strong stimulants of gastric acid secretion. (Jayaruji, 1980)

7. 7
 Non-steroidal anti-inflammatory drugs (NSAID): Most of them are associated with
ulcer formation and risk of ulcer recurrence. They damage the gastric mucosa and
precipitate upper gastrointestinal bleeding. Ulcer induced by this damage are known to be
relayed with the inhibition of Cyclooxygenase (Rainsford, 1976) that prevents
prostaglandin biosynthesis which in turn inhibits the release of mucin, a defensive factor
causing GIT damage. (Seigel, 1979)
 Genetics: The concordance for gastric ulcer is higher in monozygotic twins than in
dizygotic twins. These disease are two to three times more prevalent in first degree
relatives of affected patients than in relatives of controlled populations.
The inheritance of blood O group is associated with a modest increase in duodenal ulcer
incidence and the combined presence of blood group O and no sector status increase the
risk of developing with general population. (Cowan, 1973)

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2.3 Gastric PhysiologyRelatedto Peptic Ulcer
The formation of this disease is best explained in the interplay and relative balance of aggressive
and defensive factors.
Figure 1: Diagrammatic representation of the gastric physiology relating to peptic ulcer.
Genetics
Stress
Diet and Stroke
Excess Acid
Peptic Acid
Impaired mucosal defense
H.pylori
Peptic ulcer
NSAID

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 Acid-peptic Pathogenesis: The gastric mucosa possess the capacity to secrete acid. The
parietal cells (oxyntic cells), found in the mucosal glands of the body and fundus of the
stomach secrets HCL by a process involving oxidative phosphorylation which occurs in
the sodium potassium pump and hydrogen potassium pump.
A typical adult human stomach will secrete about 1.5 liters of gastric acid daily. Gastric
acid secretion happens in several steps. Chloride and hydrogen ions are secreted
separately from the cytoplasm of parietal cells and mixed in the canaliculated. Gastric
acid is then secreted into the lumen of the gastric gland and gradually reaches the main
stomach lumen (Dworken, 2016). The exact manner in which the secreted acid reaches
the stomach lumen is controversial, as acid must first cross the relatively pH neutral
gastric mucus layer (Dworken, 2016)
Chloride and sodium ions are secreted actively from the cytoplasm of the parietal cell
into the lumen of the canaliculus.
Parietal cells secretes hydrogen ions at three million times (3×106) more than that found
in the blood.
Each secreted hydrogen ion is accompanied by a chloride ion. With an increase in
hydrogen ion secretion, there is a reciprocal decrease in sodium ion secretion.
Bicarbonate is released from carbonic acid generated from carbon dioxide by parietal cell
carbonic anhydrase. The final step in the hydrogen ion secretion is accomplished by a
proton pump mechanism, involving a specific hydrogen-potassium adenosine
triphosphate located in the apical microvillus membrane and tubulovesicular apparatus of
the parietal cell. This pump exchanges hydrogen for potassium across the membrane.

10. 10
Multiple chemicals, neural and hormonal factors participate in regulation of gastric acid
secretion.
Gastrin and histamine- hormonal factors
Acetylcholine- neural factors.
Regulation of gastric acid secretion has been classified into 3 phases:
 Cephalic phase (eating, taste, smell and thought of food. Occurs in the brain, and its
30%) (Harvey, 2006)
 Gastric phase (distention of the stomach and presence of amino acid in the stomach.
60%)
 Intestinal phase (Distention of the small intestine and presence of amino acid in the
small intestine. 10%)
The proteolytic effects of pepsin in contact with the corrosive account for the tissue
injury that produces ulcer. Gastric acid catalyze the cleavages of inactive pepsinogen
converting then to proteolytic active pepsins and also provides the appropriate low pH
required for pepsin activity.
 Mucosal Defense: The mechanism in which the normal stomach and duodenum resist the
corrosive effect of acid -pepsin. A lot of factors contribute to this and are considered to
comprise mucosal defense.
Gastric mucin is important in mucosal defense and preventing ulceration, it is secreted by
mucus cells of the gastric mucosal epithelium gastric glands.
Bicarbonate ions, secreted by non-parietal gastric epithelial cells enter the mucus gel, and
neutralizing the hydrochloric acid/acidity of the gastric acid.
Prostaglandins can also be found in the mucus gel. (Isenberz, 1995)

11. 11
 Helicobacter pylori infection: It is a common type of bacteria that grows in the
digestive tract and has a tendency to attack the stomach lining. They are adapted to live in
the harsh acidic environment around them and reduce its acidity so they can survive. The
spiral shape of the bacteria allows them to penetrate your stomach lining, where they are
protected by mucus and your body’s immune cells are not able to reach them, they do this
by interfering with your immune response and ensure that they’re not destroyed. (Helen,
2017). Gastric colonization with Helicobacter pyloric has been reported in patients (75-
86%) with gastric ulcer syndrome. (Price, 1985)
 Non-Steroidal Anti-Inflammatory Drugs: They can cause damage to the
gastroduodenal mucosa via several mechanism, including the tropical irritant effect of
these drugs on the epithelium, impairment of the barrier properties of the mucosa,
superficial injury. The presence of acid in the lumen of the stomach also contributes to
the pathogenesis of NSAID-induced ulcers and bleeding. (Wallace, 2000)

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2.4 Diagnosis ofGastric Ulcer
Peptic ulcers are always suspected in patients with persistent dyspepsia (bloating, belching and
abdominal pain). A number of steps are to make an accurate diagnosis of ulcers.
Medical and Family History
The doctor will ask for a thorough report of a patient's dyspepsia and other important symptoms,
such as weight loss or fatigue, present and past medication use (especially chronic use of
NSAIDs), family members with ulcers and drinking and smoking habits
Ruling out Other Disorders
In addition to peptic ulcers, a number of conditions, notably gastro esophageal reflux disease
and irritable bowel syndrome, causes dyspepsia. Often however, no cause can be determined. In
such cases, the symptoms are referred collectively as functional dyspepsia. Peptic ulcer
symptoms, particularly abdominal pain and chest pain, may resemble those of the other
conditions, such as gallstones or heart attack. Certain features may help to distinguish these
different conditions. However, symptoms often overlap and it is impossible to make a diagnosis
based on symptoms alone. A number of tests are needed
Noninvasive Tests for Gastrointestinal (GI) Bleeding
When ulcers are suspected, the doctor will order tests to detect bleeding. These may include a
rectal exam, a complete blood count and a fecal occult blood test (FOBT). The FOBT tests for
hidden (occult) blood in stools. Typically, the patient is asked to supply up to 6 stool specimens

13. 13
in a specially prepared package. A small quantity of feces is smeared on treated paper, which
reacts to hydrogen peroxide. If blood is present, the paper turns blue.
Noninvasive Screening Tests for Helicobacter pylori
Simple blood, breath and stool tests can now detect Helicobacter pylori with a fairly high degree
of accuracy.
Tests for diagnosing Helicobacter pylori.
The following tests are used to diagnose Helicobacter pylori infection.
Testing may also be done after treatment to ensure the bacteria are fully eradicated.
1) Breath Test. A simple test called the carbon isotope-urea breath test (UBT) can identify up to
99% of people who harbor Helicobacter pylori. Up to 2 weeks before the test, the patient must
discontinue taking any antibiotics, bismuth-containing agents such as Pepto-Bismol and proton-
pump inhibitors (PPIs). As part of the test, the patient swallows a special substance containing
urea (a compound in mammals metabolized from nitrogen) that has been treated with carbon
atoms. If Helicobacter pylori are present, the bacteria convert the urea into carbon dioxide, which
is detected and recorded in the patient's exhaled breath after 10 minutes. This test can also be
used to confirm that Helicobacter pylori have been fully treated.
2) Blood Tests. Blood tests are used to measure antibodies to Helicobacter pylori, with results
available in minutes. Diagnostic accuracy is reported at 80 - 90%. One such important test is
called enzyme-linked immunosorbent assay (ELISA). An ELISA test of the urine is also showing
promise in children.

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3) Stool Test. A test to detect genetic fingerprints of Helicobacter pylori in the feces appears to
be as accurate as the breath test for initial detection of the bacteria and for detecting recurrences
after antibiotic therapy. This test can also be used to confirm that the Helicobacter pylori
infection has been fully treated.
4) Tissue biopsy. The most accurate way to identify the presence of Helicobacter pylori is a
tissue biopsy from the lining of the stomach. However, this is clearly an invasive task and many
patients are treated for Helicobacter pylori based on the above three noninvasive tests. It should
be noted that such tests are not as accurate as endoscopy, an invasive procedure, which is needed
to confirm a diagnosis of Helicobacter pylori. The breath and stool tests, however, can be
particularly useful after treatment to determine if a patient has been cured. If symptoms persist,
endoscopy is usually performed. Though it is an invasive procedure, it is the only procedure in
which a biopsy of stomach tissue can be taken, making it the most accurate test.
Endoscopy
Endoscopy is a procedure used to evaluate the esophagus, stomach and duodenum using an
endoscope -- a long, thin tube equipped with a tiny video camera. When combined with a biopsy,
endoscopy is the most accurate procedure for detecting the presence of peptic ulcers, bleeding
and stomach cancer or for confirming the presence of Helicobacter pylori.

15. 15
2.5 Treatment of gastric ulcer and its adverse effects.
The goal of treatment is to relieve pain, heal ulcer and prevent reoccurrence and complications.
The treatment of ulcer can often be designed either singly or in combination by:
1. Antibiotics medications to kill H.pylori: The antibiotics include amoxicillin,
clarithromycin, metronidazole, tinidazole, tetracycline and levoflaein.
2. Medications that block acid production and promote healing.
 Proton pump inhibitors: Omeprazole a substituted Benzimidazole is a potent
inhibitor of basal and stimulated gastric acid secretion in animal and man. As an acid
inhibiting drug, it inhibits the H+ transporting enzyme, H+/K+ -ATPase (Larsson,
1983). An acidic environment is needed to generate the active form of omeprazole.
Though the other new benzimidaxole dedicated such as lansoprazole (Barradel,
1992), rebeprazole (Prakash, 1996), and pantoprazole (Fitton, 1996) provide effective
symptom relief and healing of peptic ulcer.
3. Medications to reduce acid production:
 Anticholinergic: Muscarinic cholinergic antagonists can reduce basal secretion of
gastric acid by 40-50%, stimulated secretion inhibited to a lesser extent. Both non
selective muscarinic antagonists, such as atropine oxide and selective muscarinic
antagonist such as pirezepire have been used in the treatment of pelvic ulcer
disorders. (Feldman, 1984)
 H2 Receptor Antagonists: These agents selectively and reversibly bind H-2
receptors on the basolateral membrane of the parietal cell, thereby inhibiting the

16. 16
potent acid stimulating effects of histamine, Cimetidine (Baron, 1981), a first clinical
drug followed by more potent ranitidine (Riley,1982), famotidire (iampoli,1986),
roxatidine (Freson, 1990) etc.
4. Antacids that neutralize stomach acid:
 Antacid: These drugs have been the mainstay of gastric ulcer therapy. Their efficacy
in neutralizing acids has been proven in controlled studies. Magnesium and
aluminum hydroxides are the active ingredients in most antacids, besides these
sodium and calcium compounds are available. They are used in combination for the
treatment. Moderate-large doses of antacids may be comparable to H-2 receptor
antagonists in healing gastric ulcer, but they are associated with high incidence of
side effect. (Berstad, 1986)
5. Medications that protect the lining of stomach and small intestine: In some cases the
doctor may prescribe medication called cytoprotecrive agents that help protect the tissue
that line the stomach. Options include sucralfate and misoprostol.

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2.6 Moringa:Introduction.
Moringa oleifera as discussed earlier is a widely cultivated species of Moringa, from the family
of Moringacease. It is found mostly in the Southern foothills of the Himalayas in northwestern
India. It is widely cultivated in tropical and subtropical areas all over Asia, Africa and South
America.
But the Moringa powder made from naturally dries Moringa leaves is what is going to be used in
the course of this experiment. It has a delicious spinachy green flavor and contains a lot of
nutrients making it have a variety of benefits including being a major source of antioxidant.
2.7 History and other research work done with Moringa.
Uses of natural herbal remedies in gastric ulcer have been prevalent in folk medicine in different
countries. Seeds, leaves, oil, sap, bark, roots, and flowers are widely used in traditional medicine.
Moringa leaves have been characterized to contain a desirable nutritional balance, containing
vitamins, minerals, amino acids, and fatty acids (Moyo et al., 2011; Teixeira et al., 2014; Razis et
al., 2014). Additionally, the leaves are reported to contain various types of antioxidant
compounds such as ascorbic acid, flavonoids, phenolics, and carotenoids (Alhakmani et al.,
2013; Vongsak et al., 2014). According to several commentaries (Anwar et al., 2007; Mbikay,
2012; Razis et al., 2014), various preparations of M. oleifera are used for their anti-inflammatory,
antihypertensive, diuretic, antimicrobial, antioxidant, antidiabetic, antihyperlipidemic,
antineoplastic, antipyretic, antiulcer, cardioprotectant, and hepatoprotectant activities. The
therapeutic potential of M. oleifera leaves in treating hyperglycemia and dyslipidemia was
reviewed by Mbikay (2012). Razis et al. (2014) summarized potential health benefits of

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M. oleifera, focusing on their nutritional content as well as antioxidant and antimicrobial
characteristics. The effect of a hexane extract of M. oleifera leaves on reproductive organs of
male rats was examined (Cajuday and Pocsidio, 2010)
Kumari (2010) treated type 2 diabetic subjects with 8 g of powdered M. oleifera leaf in a tablet
form per day for 40 days. Bakre et al. (2013) have shown an ethanol extract of M. oleifera leaves
possesses Central nervous System (CNS) depressant and anticonvulsant activities in mice
through the enhancement of central inhibitory mechanism involving release of γ-amino butyric
acid.
Jung (2014) has shown that an aqueous extract of M. oleifera leaves exhibited significant
antineoplastic activity against a lung cancer cell line and several other types of cancer cells.
Tende et al. (2011) examined the effects of an ethanol extract of M. oleifera leaves on blood
glucose levels of streptozotocin-induced diabetic rats
Yassa and Tohamy (2014) have also assessed the antidiabetic and antioxidant potential of an
aqueous extract of M. oleifera leaves in streptozotocin-induced diabetic rats.
2.7.1 Moringa and Anti-oxidant effect
Moringa is a rich source of antioxidant. Its leaves act as a good source of natural antioxidant due
to the presence of various types of antioxidant compounds such as ascorbic acid, flavonoids,
phenolics and carotenoids (Anwar, 2007). Moringa is said to have approximately 46 antioxidants
and is one of the most powerful sources of natural anti-oxidants. Anti-oxidants supply the free
atoms needed by the human body and mitigate the effect of free radicals. Moringa leaves are rich
in Flavonoids, a class of anti-oxidants. The beta carotene present in Moringa leaves also acts as

19. 19
antioxidants (Mishra, 2011). The antioxidants will have the maximum impact on the damage
causing free radicals, only when it is ingested in combination with nutrients and a group of
antioxidants. A combination of antioxidants is more effective than a single antioxidant on an
equal weight basis due to antioxidant cascade mechanism. It has been reported that aqueous
extracts of leaf, fruit and seed of Moringa act as an antioxidant. During a study reporting
antioxidant property of freeze dried Moringa leaves from different extraction procedures,
2.7.2 Other uses and benefits of Moringa.
The Moringa powder gotten from the dried leaves would be used for this experiment. This
powder are nutritional powerhouse that a wide range of nutrients such as:
 25% plant protein including all 9 essential amino acids
 24% fiber
 A rich/significant amount of vitamins, such as vitamin A, vitamin B1, B2, B3, B6, folate
and vitamin C (ascorbic acid)
 Minerals such as calcium, potassium, iron, magnesium, phosphorus and zinc.
 It has low amount of fats and no cholesterol
 And last but not the least it is high in antioxidants.
But in the course of this experiment, we would be more focused on the antioxidant effects of the
Moringa Oleifera.
It treats edema, protects the liver, treats stomach disorder, skin care and hair care, acts as
antibacterial agents, cures cancer, treats neurodegenerative diseases, improves bone health, helps

20. 20
in muscle growth, boost immunity, reduces tiredness and fatigue, protects cardiovascular system,
treats diabetics and asthma, protects against kidney problems, has anti-fertility effects, heals
wounds, reduces hypertension, improves eye health, treats anemia and sickle cell and on and on
the list goes for the variety of the benefits this powder provides to human health. But our main
focus in this study is its effect on gastric ulceration.
2.8 Oxidative stress
This simply the imbalance between the systemic manifestation of reactive oxygen species (free
radicals) and the ability of the body to detoxify their harmful effects through neutralization by
antioxidants. So the antioxidant capacity is simply overwhelmed.
So this prevents the antioxidant from repairing the resulting damage. Disturbances in the normal
redox state of cells can cause toxic effects through the production of perioxides and free radicals
that damages all component of the cell including proteins, lipids and DNA. Oxidative stress from
oxidative metabolism causes base damage, as well as strand breaks in DNA. Base damage is
mostly indirect and caused by reactive oxygen species (ROS) generated e.g. superoxide radicals,
hydroxyl radicals and hydrogen perioxides. (Chandra, 2015)
2.8.1 Damage caused by oxidative stress
It may lead to many path physical conditions in the body. Some neurodegenerative diseases such
as Parkinson's disease and Alzheimer's disease, gene mutations and cancer, chronic fatigue
syndrome, fragile X syndrome, heart and blood vessel disorders, arteriosclerosis, heart failure,

21. 21
heart attack and inflammatory disease and even according to this study ulcer though Helicobacter
pylori which increases reactive oxygen and nitrogen species in human stomach. (Ananya, 2017)
2.9 Free Radicals.
A radical in chemistry is an atom, molecule, or in that has an unpaired valence electron (Hayyan,
2016). Free radicals on the other hand are atoms or groups of atoms with an odd (unpaired)
number of electrons and can be formed when oxygen interacts with certain molecules. They are
unstable molecules that damage cells and contribute to aging and diseases. They are highly
reactive with other molecules.
Oxygen by products by nature are relatively unreactive molecules but some of these can undergo
metabolism within the biological system and give rise to the highly reactive oxidants. Not all of
the reactive oxygen species are harmful to the body. Some of them are useful in killing
pathogens and microbes.
But most of them chemically react with cell component such as DNA, protein or lipid and steal
their electrons in order to stabilize, thereby destabilizing the cell component molecule which in
turn seek and steal electron from another molecule, therefore triggering a large free radical chain
reaction. (Ananya, 2017)

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2.9.1 Antioxidant
Every cell that utilizes enzymes and oxygen to perform functions is exposed to oxygen free
radical reactions that have the potential to cause serious damage to cell.
Antioxidant is a molecule that inhibits the oxidation of other molecules. They do this by donating
an electron to the free radicals (reactive oxygen) without becoming destabilized themselves. An
imbalance between the oxidants and antioxidants is the underlying basis of oxidative stress. They
can be found in many foods, including fruits and vegetables. That is why this study would like to
prove the antioxidant property in Moringa in the treatment of gastric ulceration. (Ananya, 2017)
2.9.2 The pH of the stomach
The stomach secretes hydrochloric acid, but the pH of the stomach isn't necessarily the same as
the pH of the acid. The pH of the stomach varies, from 1-2 up to 4-5. After eating, the stomach
releases enzymes called proteases as well as hydrochloric acid to aid in digestion. By itself, the
acid doesn't really do much for digestion, but the proteases that cleave proteins work best in an
acidic environment or low pH, so after a high-protein meal, your stomach pH may drop to as low
as 1 or 2.
However, buffers quickly raise the pH back to 3 or 4. After the meal has been digested, the
stomach pH returns to a resting level of about 4 or 5. The stomach secretes acid in response to
food, so first thing in the morning you can expect a slightly acidic stomach pH, but not an acidic
level representative of pure hydrochloric acid.

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Chemical Composition of Gastric Juice
The liquid inside the stomach is called gastric juice. It isn't just acid and enzymes, but a complex
mixture of several chemicals. Take a look at the molecules, the cells that make them, and the
function of the different components:
Water - Water doesn't affect the pH of the stomach, but it does serve to provide enough liquidity
that food, enzymes, and acids can readily mix together. Some enzymes require water in order to
function.
Mucous - Mucous (or mucus) is produced by cells in the mouth, esophagus, and stomach. It
eases the passage of food through the gastrointestinal tract and protects the lining of the stomach
from being attacked by acid. Neck cells also secrete bicarbonate, which buffers the acid and
controls pH.
Hydrochloric Acid - This potent acid is secreted by the parietal cells of the stomach. It kills
bacteria and other potential pathogens in food and converts the enzyme pepsinogen into pepsin,
which breaks secondary and tertiary proteins into smaller, more easily digested molecules.
Pepsinogen - Pepsinogen is secreted by chief cells in the stomach. Once it's activated by low pH
it is converted to pepsin, which then helps in the digestion of proteins.
Hormones and Electrolytes - Gastric juice also contains hormones and electrolytes, which aid in
organ function, food digestion, and nutrient absorption. The enteroendocrine cells secrete
multiple hormones.
Gastric Lipase - This is an enzyme made by chief cells in the stomach that aids in breaking up
short-chain and medium-chain fats.

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Intrinsic Factor - The parietal cells of the stomach secrete intrinsic factor, which is necessary for
vitamin B-12 absorption.
Amylase - Amylase is an enzyme found primarily in saliva, where it acts to break down
carbohydrates. It's found in the stomach because you swallow saliva as well as food, but it is
inactivated by the low pH. Additional amylase is secreted into the small intestine.
The mechanical churning action of the stomach mixes everything together to form what is called
chyme. Eventually, chyme leaves the stomach and processed to the small intestine so that the
acid can be neutralized, digestion can proceed, and nutrients may be absorbed. (Helmenstine,
2017)

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CHAPTER THREE
3.0 MATERIALS AND METHODS
3.1.0 Materials
100-120g 20 male Wistar Rats
Indomethacin
Moringa Oleifera
Pellet feed (1 bag)
Water
Ready-made cages (4)
Oral cannula
Dissecting sets
Plain and EDTA bottles
Hand gloves
Oxidative stress protocol
Capillary tubes
Ulcer score microscope
Sodium potassium protocol
pH meter

26. 26
Spectrometer
Centrifuge
Normal saline.
3.2.0 Animals: Experimental animals for study
Twenty male wistar rats were weighed at about 100-120g, and were kept at the Babcock
University Animal house. They were fed with pellet feed and water throughout the period of the
study.
They were first left to gain acclimatization for about 5-7 days in a sterile environment with
standard light, temperature and humidity.
3.3. Grouping of animals and treatment.
Twenty healthy adult Male Wistar rats were randomly divided into 4 different groups in a very
tidy environment with standard light, temperature and humidity. The rats would be left to gain
acclimatization 3-5 days before the commencement of the experiment.
The first five rats (Group 1- normal control) received pellet feed and water only daily. The rats in
Group 2 (ulcerated control) were given indomethacin to induce ulcer. Group 3 of the rats
received Moringa and normal pellet feed with water. And the last Group 4 were given both
Moringa and indomethacin. The dose of Moringa given to the rats was 50mg/kg body weight.

27. 27
While the dose of indomethacin given 30mg/kg body weight. Both the Moringa and
indomethacin were administered orally. The Moringa was administered once daily throughout
the experimental period (10-14 days).
Table1: Showing the animal groupings and treatment
Group 1 5 Control group (no treatment)
Group 2 5 Ulcerated control- indomethacin and feed
Group 3 5 Administration of Moringa only
Group 4 5 Administration of Moringa and indomethacin (ulcer)
3.4 Ulcer induction using indomethacin
Gastric ulceration was induced in the rats from Group 2 and Group 4. Briefly the rats where
administered with a single oral dose of indomethacin (30mg/kg body weight). They were
deprived of food 24 hours prior to ulcer induction but were supplied with water, their beddings
were also cleared so as to avoid the rat consuming it.
Various degrees of ulceration manifested 4 hours after indomethacin administration.

28. 28
3.5 Isolationof stomach and collectionof gastric juice/content.
4 hours post ulcer induction 3 rats (head, back and tail) each from all the 4 groups where
humanely sacrificed by cervical dislocation.
The abdomen was opened and a small incision was made transversely on the esophagus. A small
tube connected to an oral cannula filled with a solution of normal saline solution was pumped
into the stomach. The whole content of the stomach including the saline was retrieved back using
the cannula and emptied into tubes.
All this were done for all the 12 rats from each of the different groups.
Note: The base of the stomach/beginning of the small intestine was clipped in order to avoid the
flow of the saline into the intestine and out if the body. The stomach was then cut out and put in
the freezer.
3.6 Determinant of the pH of the gastric content.
After the gastric content were collected from each of the 12 rats from all 4 groups. Their pH was
checked using a pH meter. Each of the tube containing the gastric contents for the different was
left for about 1-2minutes for the pH reading to stabilize.

29. 29
3.7. Quantification of the ulceration (Ulcer score)
The stomach of each of the 12 rats from the 4 groups (3 rats from each group) were gotten from
the freezer. The stomach was incised along the greater curvature and washed/cleaned with
distilled water, then one by one the stomachs were spread on a microscope slide and ulceration
was scored.
3.7.1. Scoring of ulcer was made as follows
Normal stomach…….. (0)
Red coloration………. (0.5)
Spot ulcer…………… (1)
Hemorrhagic streak…. (1.5)
Ulcers………………... (2)
Perforation………….... (3)
3.7.2 Calculation of ulcer index
UI= Ulcer Index
UI= (UN+US+UP) ×10-1
UN= Average of number of ulcer per animal
US= Average of severity score
UP= Percentage of animal with ulcer (Vogel, 2002)

30. 30
3.8 Preparationof stomachhomogenate and Assay of Na+
/K+
ATPase specific
activity.
The 12 samples of rats’ stomach were obtained from the freezer where they were stored. Each of
the stomach samples was weighed and 0.3g of each of the 12 samples was taken. The 0.3g of
each samples were added to 3ml of phosphate buffer. The sample and the buffer were both
homogenized using both the manual and the electrical homogenizer.
Synaptosomal Na+/K+ ATPase activity was assayed in reaction mixtures of (i) 30mm imidazole-
HCL, 130 mm NaCL, 20mm KCL, 4 mm MgC12, and (ii) 30 mm imidazole-HCL, 4 mm MgCL,
and 1 ouabain, at pH 7.4, each containing 20D50 µg Synaptosomal protein. For complete
ouabain binding, the tubes were pre-incubated for 60 min in ice and dark. The reaction was
started by adding 4mm Tris-ATP (Tris hydroxymethylaminomethane salt of adenosine
triphosphate, Sigma Co.) and incubated at 37 ∞C for 10 min. The total volume of the reaction
mixture was 1 ml. The enzyme activity was stopped by addition of 100 µl of 20% sodiumdodec
ylsulfate. The Pi formed was determined from the reaction mixture (Baginski et al., 1967).
Na+/K+- ATPase activity was calculated as difference in the Pi content in nmol/min between
media (i) and (ii). Enzyme protein was solubilized using 100 µl of 20% SDS and from this
mixture and aliquot (0.2 ml from a total volume of 1.1ml) was taken for Pi estimation followed
by a quick addition of 0.5 ml of reagent A (containing 3% ascorbic acid in 0.5 and HCl and 0.5%
ammonium molybddate solution) in a total volume of 1 ml and the tubes were kept in ice for 10
min. Then 1 ml of reagent B (containing 2% sodium eta-arsenite, 2% trisodium citrate and 2%
acetate acid) was added. The color developed after 10 min at 37 ∞C was read at a wavelength
850 nm in a Beckman spectrophotometer DU-70 (Esmann, 1988)

31. 31
3.9 Assayof antioxidant indices/oxidative stress
The following antioxidant enzymes activities will be determined spectrometrically as follows:
3.9.1 Determination of Superoxide Dismutase (SOD) activity
Superoxide Dismutase activity was determined by its ability to inhibit the auto-oxidation of
epinephrine determined by the increase in absorbance at 480nm as described by Sun and Zigma
(1978). The reaction mixture (3 ml) contained 2.95 ml 0.05 M sodium carbonate buffer pH 10.2,
0.02 ml of liver homogenate and 0.03 ml of epinephrine in 0.005 N HCL was used to initiate the
reaction. The reference cuvette contained 2.95 ml buffer, 0.03 ml of substrate (epinephrine)
0.02 ml of water. Enzyme activity was calculated by measuring the change in absorbance at 480
nm for 5 min= 4020M-1 cm-1
3.9.2 Determination of Catalase activity
Serum catalase activity was determined according to the method of Beers and Sizer as described
by Usoh et al., (2005) by measuring the decrease in absorbance at 240nm due to the
decomposition in a H2O2 UV recording spectrophotometer. The reaction mixture (3 ml)
contained 0.1 ml of serum in phosphate buffer (50 mM, pH 7.0) and 2.9 ml of 30 mM H2O2 in
phosphate buffer pH 7.0. An extinction coefficient for at 240nm H2O20 of 40.0 M-1cm-1 (Aebi
1984) was used for the calculation. The specific activity of catalase was expressed as moles of
H2O2 reduced per minute per mg protein.

32. 32
3.9.3 Reduced Glutathione Determination
The reduced glutathione (GSH) content of liver tissue as non-protein sulphydryls was estimated
according to the method described by Sedlak and Lindsay (1968). To the homogenate 10% TCA
was added, centrifuged. 1.0ml of supernatant was treated with 0.5ml of Elman’s reagent (19.8mg
of 5, 5-dithiobisnitro benzoic acid (DTNB) in 100ml of 0.1% sodium nitrate) and 3.0ml of
phosphate buffer (0.2M, pH 8.0). The absorbance was read at 412nm. ? = 1.34 x 104 M-1 cm-1
3.9.4 Lipid Peroxidation
Malondialdehyde (MDA) an index of lipid peroxidation was determined using the method of
Buege and Aust (1978). 1.0 ml of the supernatant was added to 2 ml of (1:1:1 ratio) TCA-TBA-
HCl reagent (thiobarbituric acid 0.37%, 0.24N HCl and 15% TCA) tricarboxylic acid-
thiobarbituric acid-hydrochloric acid reagent boiled at 100oC for 15 min, and allowed to cool.
Flocculent materials were removed by centrifuging at 3000 rpm for 10 min. The supernatant was
removed and the absorbance read at 532 nm against a blank. MDA was calculated using the
molar extinction coefficient for MDATBA- complex of 1.56 × 105 M-1CM-1.

33. 33
3.10 STATISTICALANALYSIS
Data would be expressed as mean ± standard error of mean (SEM) and analyzed with instant
graph pad software sing student’s test for comparisons between two groups and one way
ANOVA for multiple comparisons. Differences between groups will be considered significant at
P˂0.05

34. 34
CHAPTER FOUR
RESULTS
Table 2: gastric pH in control and test groups over the period of 2 weeks.
Group 1(control)
Group
2(Ulcer)
Group
3(Moringa)
Group 4(Ulcer and
Moringa)
Gastric
pH 3.41±0.04 2.5±0.20 * 3.71±0.19 # 3.97±0.21 #
*=p˂0.05 when comparing with the control group (group 1)
#=p˂0.05 when comparing with the ulcerated group (group 2)
Table 2 above shows the result for the pH of the gastric content that was collected in the control
and the test groups. The mean pH recorded for the control group was 3.41±0.04 while that for the
ulcer group was significantly lower (p<0.05) at 2.5±0.20 when compared with the control group.
The recorded mean pH for the Moringa group and ulcer and Moringa treated groups were
slightly higher at 3.71±0.19 and 3.97±0.21 respectively when compared with the control group

35. 35
Group
1(control)
Group
2(Ulcer)
Group
3(Moringa)
Group
4(Ulcer and
Moringa)
0
1
2
3
4
5
*
#
#
pH
Figure 2: gastric pH in the control and test groups over the study of 2 week.
*=p˂0.05 when comparing with the control group (group 1)
#=p˂0.05 when comparing with the ulcerated group (group 2)

36. 36
Table 3: ulcer score in the control and test groups over the study period of 2 week.
Control Ulcer Moringa
Moringa +
Ulcer
Ulcer score 0.03±0.03 2.17±0.55* 0.33±0.19# 0.42±0.16#
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)
Table 3 above shows the result for the ulcer score of the stomach that done for the control and
the test groups. The mean ulcer score recorded for the control group was 0.03±0.03 while that for
the ulcer group was significantly higher (p<0.05) at 2.17±0.55 when compared with the control
group. The recorded mean pH for the Moringa group and Moringa and ulcer treated groups were
slightly higher at 0.33±0.19 and 0.42±0.16 respectively when compared with the control group

37. 37
Ulcer Score
C
ontrol
U
lcer
M
oringa
M
oringa
+
U
lcer
0
1
2
3
*
# #
Figure 3: ulcer score in the control and test groups over the study period of 2 week.
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)

38. 38
Table 4: Sodium potassium ATPase pump in the control and test groups over the study period of
2 week.
control Ulcer Moringa Moringa and
ulcer
Sodium
potassium
ATPase pump 1.10±0.03 1.02±0.02 1.21±0.02 1.08±0.04
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)
Table 4 above shows the result for the sodium potassium ATPase pump test of the stomach that
done for the control and the test groups. The mean score recorded for the control group was
1.10±0.03 while that for the ulcer group was significantly lower (p<0.05) at 1.02±0.02 when
compared with the control group. The recorded mean for the Moringa group slightly higher at
1.21±0.02 and Moringa and ulcer treated groups was slightly lower and 1.08±0.04 respectively
when compared with the control group.

39. 39
Sodium potassium ATPase
C
ontrol
U
lcer
M
oringa
U
lcer +
M
oringa
1.1
1.2
1.3
*#
mg/ml
Figure 4: Na+/K+ ATPase test in the control and test groups over the study period of 2 week.
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)

40. 40
Assay of anti-oxidant/oxidative stress
Table 5: Glutathione test in the control and test groups over the study period of 2 week.
Control Ulcer Moringa Moringa and
ulcer
Glutathione
µmol/ml 5.52±0.20 3.56±0.64 * 5.67±0.15 # 5.01±0.40 #
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)
Table 5 above shows the result for the glutathione test done in the stomach for the control and
the test groups. The mean score recorded for the control group was 5.52±0.20 while that for the
ulcer group was significantly lower (p<0.05) at 3.56±0.64 when compared with the control
group. The recorded mean for the Moringa group was slightly higher at 5.67±0.15 and Moringa
and ulcer treated groups was significantly lower at 5.01±0.40 when compared with the control
group.

41. 41
Figure 5: Glutathione test in the control and test groups over the study period of 2 week.
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)
Glutathione
C
ontrol
U
lcer
m
oringa
M
oringa
+
U
lcer
0
2
4
6
8
*
# #
µmol/ml

42. 42
Table 6: Superoxide dismutase test in the control and test groups over the study period of 2
week.
control Ulcer Moringa Moringa and
ulcer
Superoxide
dismutase
Mg/ml 6.98±0.29 5.18±0.29 * 6.72±0.35 # 5.87±0.30 *
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)
Table 6 above shows the result for the Superoxide dismutase done for the control and the test
groups. The score recorded for the control group was 6.98±0.29 while that for the ulcer group
was significantly lower (p<0.05) at 5.18±0.29 when compared with the control group. The
recorded mean for the Moringa group and Moringa and ulcer treated groups were slightly lower
at 6.72±0.35 and 5.87±0.30 respectively when compared with the control group

43. 43
superoxide dismutase
control
ulcer
M
oringa
M
oringa
and
ulcer
0
2
4
6
8
* *
#
mg/ml
Figure 6: Superoxide dismutase in the control and test groups over the study period of 2 week.
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)

44. 44
Table 7: Catalase test in the control and test groups over the study period of 2 week
Control Ulcer Moringa Moringa and
ulcer
Catalase
Mg/ml 18.44±1.25 3.57±1.80 15.28±1.49 11.06±1.76
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)
Table 7 above shows the result for the catalase test done for the control group and the test
groups. The mean score recorded for the control group was 18.44±1.25 while that for the ulcer
group was significantly lower (p<0.05) at 3.57±1.80 when compared with the control group. The
recorded mean for the Moringa group and Moringa and ulcer treated groups were slightly lower
at 15.28±1.49 and 11.06±1.76 respectively when compared with the control group

45. 45
Catalase
G
roup
1(controlgroup)G
roup
2(U
lcer)
G
roup
3(M
oringa)
G
roup
4(U
lcerand
M
oringa)
0
5
10
15
20
25
*#
*
#
mg/ml
Figure 7: catalase test in the control and test groups over the study period of 2 week.
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)

46. 46
Table 8: Lipid peroxidation test in the control and test groups over the study period of 2 week
control Ulcer Moringa Moringa and
ulcer
Lipid
peroxidation
nmol/ml 4.13±0.22 7.71±0.46 * 3.81±0.19 # 5.59±0.68 *#@
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)
Table 8 above shows the result for the Lipid peroxidation test done for the control and the test
groups. The score recorded for the control group was 4.13±0.22 while that for the ulcer group
was significantly higher (p<0.05) at 7.71±0.46 when compared with the control group. The
recorded mean for the Moringa group was slightly lower at 3.81±0.19 and Moringa and ulcer
treated groups were slightly higher at 5.59±0.68 respectively when compared with the control
group.

47. 47
Lipid perioxidation
C
ontrol
U
lcer
M
oringa
M
oringa
+
U
lcer
0
2
4
6
8
10
*
*#@
#
nmol/ml
Figure 8: Lipid peroxidation in the control and test groups over the study period of 2 week.
*=p˂0.05 when comparing to control group (group 1)
#=p˂0.05 when comparing to ulcerated group (group 2)
@=p<0.05 when comparing to Moringa group (group 3)

48. 48
CHAPTER FIVE
5.0 DISSCUSSION AND CONCLUSION
5.1 DISCUSSION
Gastric ulceration occurs when gastric acid is produced in excess, leading to the damage of the
mucosal membrane enabling the Helicobacter pylori to degenerate the barrier and cause internal
infection or it would expose the mucosal epithelial to more action of hydrochloric acid resulting
in sores on the membrane from where the acid has come in contact with it. Or gastric ulceration
can also be caused by the action of Non-steroidal inflammatory drugs. So in summary gastric
ulceration is caused by the imbalance between aggressive factors (principally secreted gastric
acid and pepsin) and factors that comprise mucosal defense or mucosal resistance to ulceration.
Gastric ulcer may occur when the aggressive effect of acid and pepsin outweighs the protective
effect of gastric mucosal resistance.
Indomethacin was able to induce ulcer in the rats because it is a non-steroidal anti-inflammatory
drugs. NSAIDs causes’ damage to the gastric mucosa through various mechanisms, including
damage to the gastric mucosa and precipitate upper gastrointestinal bleeding, this happens
because of the irritant effect of the drug on the epithelium, reduction of gastric mucosal blood
flow and interference with the repair of superficial injury. Impairment of the barrier properties of
the mucosa by suppression of gastric prostaglandin synthesis (Baillieres, 2000) damage are
known to be relayed with the inhibition of Cyclooxygenase (Rainsford, 1976) that prevents
prostaglandin biosynthesis which in turn inhibits the release of mucin, a defensive factor causing
GIT damage. (Seigel, 1979).

49. 49
The treatment that was used in this experiment to treat gastric ulceration was Moringa oleifera.
The commercial Moringa pack made from naturally dried leaves was used in the course of the
experiment. Moringa contains a lot of nutrients making it have a variety of benefits including
being a major source of antioxidant. Oxidative stress is the imbalance between the systemic
manifestation of reactive oxygen/free radicals and the ability of the body to detoxify it (this is the
antioxidant). Oxidative stress occurs when the activities of the body internal supply of
antioxidant is outweighed/overweighed by the free radicals. So in order to balance it out, excess
antioxidant are needed by the body. And that is when Moringa comes into place. Oxidative stress
causes the overstimulation of the sodium potassium ATPase pump leading to over secretion of
hydrochloric acids leading to gastric ulceration. So Moringa is believed to balance out the
reaction because of the excess antioxidant it produces, making it possible for the body to return
to its normal state.
So like I stated earlier the purpose of this experiment is to test the Moringa's antioxidant
capability and to prove that Moringa would be very helpful in the prevention of gastric
ulceration. So after gastric ulcer induction the following tests gastric pH, ulcer score, sodium
potassium ATPase pump and oxidative stress were carried out after 2 weeks of Moringa
administration to test the effectiveness of the Moringa treatment.
Assessing the result gotten from the pH test, it was shown that Group 3(Moringa) and Group
4(Moringa and Ulcer) had relatively normal pH (not too acidic), and their pH is almost the same
when compared to the control group (group 1), whereas when compared to Group 2(ulcerated
group) there is a significant amount of difference in the pH. Gastric ulceration was induced both
in Group 2 and Group 4. But the gastric pH if Group 4 isn't as acidic as that of Group 2, because

50. 50
of the Moringa treatment. This simply shows that Moringa was able to reduce the excess
secretion of Gastric acid, thereby increasing the gastric pH.
Also assessing the results gotten from Ulcer score. It was shown that Group 3(Moringa) and
group 4(Moringa and ulcer) weren't affected by ulceration sores, and when compared to Group 1
(Control- normal stomach) they are much alike. But when Group 3 (Moringa) and Group 4
(Moringa and Ulcer), are compared to Group 2 (ulceration stomach), you can see the ulcerated
stomach in Group 2 has a lot of sores from the ulcer. Both Group 2 and Group 4 were induced
with ulcer, but group 4 was treated with Moringa weeks before the induction. So when Group 2
and Group 4 are compared together, Group 4 has lesser or no sores unlike Group 2. Therefore
this signifies that Moringa was able to protect the stomach against Gastric Ulceration.
Sodium potassium ATPase pump is located in the cells of the stomach, and it is known to
regulate the production/secretion of gastric juice/acid or HCL secretion. Assessing result gotten
from sodium potassium ATPase test. It was shown that in Group 2 (ulcer group) the level of
sodium potassium ATPase pump is low, because of the low level of the pump gastric acid
secretion isn’t regulated appropriately leading to over production of the gastric juice which then
causes ulcer. Control group has normal level of the pump. Group 3 (Moringa) has the highest
level of pump because of the adding of more pumps to the already normal level of Na+/K+ pump.
While the level of Na+/K+ pump in the Moringa + ulcer group (Group 4) is higher than that of the
Ulcer group because of the Moringa administered. This test proves that Moringa can prevent
Ulcer because when the ulcer was induced it didn’t affect the regulation of HCL production, that
is, it maintained the normal amount/level of the pump in the stomach.
The oxidative stress test performed is meant to show how the oxidative stress affected the level
of antioxidants produced by the body. When ulcer was induced it led to the increase in oxidative

51. 51
stress in the body. Glutathione, catalase, superoxide dismutase and Lipid peroxidation are all
antioxidants produced naturally by the body. From the glutathione test, catalase test and
superoxide dismutase test done, the levels of these antioxidants in stomach of the ulcer group
(Group 2) are relatively low when compared to that of the control group because ulcer increases
the level of oxidative stress which in turn reduces/decreases the level of all these antioxidants in
stomach. Also from this 3 tests the Control group (Group 1) has the highest number because this
is the normal level of all these antioxidants in the body. While that for Group 4 is slightly lower
when compared to the control group because of the ulcer, but it is still higher when compared
with group 2. This shows/proves that the Moringa was able to prevent the ulcer because the level
of the antioxidants are relatively normal because the Moringa added more antioxidants to the
stomach, so when the ulcer was induced the oxidative stress didn’t affect the level of antioxidant
in the stomach.
But the last oxidative test done Lipid peroxide is completely different. Lipid peroxide is a
process that produces or causes the release of Malondialdehyde. Oxidative stress increases this
process, leading to the over production of Malondialdehyde which increases its level in the
stomach. So from the result Group 2 has the highest level of Malondialdehyde because the ulcer
group has the highest oxidative stress level. The level for of Malondialdehyde in Group 1 is
normal, while that for Group 4 is low when compared with Ulcer group (Group 2), this means
the level of Malondialdehyde reduced because of the administration of Moringa. So this proved
that Moringa prevented the ulcer because the induction of Ulcer didn’t affect the level of
Malondialdehyde in the stomach.

52. 52
5.2 CONCLUSION
It was concluded from this study that the Moringa Oleifera that was administered was able to
prevent gastric ulceration through its anti-oxidant effect. The anti-oxidant potential effectiveness
of the Moringa Oleifera was tested against indomethacin induced gastric ulcer in rats using the
pH, ulcer score, anti-oxidant assay and sodium potassium protocol and its effectiveness was
proven. It was also obtained from the study that Moringa increased the overall health of the
normal Wistar rats in Group 3.

53. 53
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