ANTIULCER ACTIVITY OF HEDGE MUSTARD SEEDS AGAINST ASPIRIN INDUCED ULCERS

IN RATS

1. INTRODUCTION

1.1. Background history of medicinal plants

Though the principles themselves took a long time to evolve into a body of knowledge, there has always

been a notable interest for health care and illness cures throughout human history. The understanding of

the essential principles underpinning the biochemical events leading to pharmacological actions is a

rational approach to the research of medications and their activities. Nowadays, more understanding into

drug behaviour at the macromolecular level is being generated, and there is a growing body of direct and

indirect data to back up these biological drug action hypotheses. Natural goods have a history of therapy

in the form of folk medicines in the so-called pre-scientific period, but little of today's medication therapy

is based on these cures. Some of the natural ingredients that are now being utilised, either as is or as

derivatives, were once employed for other purposes, such as arrow poisons, religious or other rites, and

even cosmetics. Opium, belladonna, cinchona bark, ergot, curare, nutmeg, calabar bean, foxglove, and

squill are examples of such products. Many substances that were once utilised as folk treatments have been

phased out. Anatomy and physiology understanding are vital in the creation of medication therapy. For

example, Nux vomica was the first medicine given to animals by Javanese arrow through multiple

pathways, causing convulsions and suffocation. For the first time, the spinal cord was removed and

sectioned to determine the location of action, and the active component, strychnine, was later discovered

and isolated. Following the French Revolution, science grew increasingly empirical, and logic-based

medications became obsolete1.

The herb known as "Ma huang," a species of Ephedra used medicinally in China for over 5000 years1, is

perhaps the first recorded usage of a medicinal plant in the "Rigveda" and one cited in current writings

(Foye et al., 1995). In the Vedic era, Ayurveda mentions the use of plants for sickness treatment. Many

diuretics, emmenagogues, carminatives, rubefacients, dermatologic treatments, expectorants,

anthelmentics, and other drugs are derived from plants. The Globe Health Organization (WHO)

attempted to identify all therapeutic plants in the world some years ago. The list contained almost 20,000

different species. The NAPRALERT database records ethnomedicinal applications for 9200 of 33000

monocots, dicots, gymnosperms, pteridophytes, bryophytes, and lichen species, implying that over a
quarter of all plants on the planet have been utilised ethnomedicinally2. Although plant-based

medications are still used for some of these purposes, synthetic pharmaceuticals now make up the majority

of the goods available. It's worth noting that much of the work on synthetic substances began after

scientists and researchers had discovered and classified active natural molecules. Plants native to a specific

location were not readily available throughout the year in the same way that they were in other parts of

the world. As a result, the creation of synthetic compounds, and therefore active ingredients, was largely

motivated by the need to secure enough supply while adhering to established quality standards. It's worth

noting that out of over 1 billion prescriptions written by pharmacies in the United States in 1967, over 243

million, or roughly 23%, contained one or more plant-based products. About 50 plant genera were

represented in the goods, which included 50 pure chemicals and 40 crude or semi-purified varieties of

plant extracts. Even now, the ratio of medicines has not altered. Plant drugs, such as those used as

immunomodulators in our traditional systems of medicine, such as Ayurveda, Siddha, and Unani,

continue to play an important role in modern medicine, particularly in grey areas where there is little or no

therapy, such as in the grey areas of modern medicine where there is little or no therapy.

Curing was largely mystical and relied on reasoning rather than empirical proof at the start of human

cultural progress. A few plants classified as magic or psychedelic plants, which are generally

hallucinogenic, were employed. Later, empirical medicine evolved, which used a variety of plants to cure a

variety of ailments. This changed substantially with Dioscorides' Materia Medica and Watt's Dictionary

of Economic Plants of India, both of which explain the qualities of different plant-based medications.

Herbal medications were widely used in ancient culture, and their usage was carefully documented in the

form of pharmacopoeia3.

India was not far behind, and thousands of years ago had a large pharmacopoeia. This experience was

methodically documented and formed the Materia Medica of ancient medical systems in various eastern

civilizations, such as India, China, and the Arab world. Traditional medicine is a wide word that refers to a

variety of historic and natural health care techniques that existed before contemporary scientific methods

were applied to health care4. Traditional plant medicinal expertise from Africa and South America, as well

as China and India, has given modern medicine numerous novel medications.

At least 120 chemical compounds originating from plants are currently regarded significant medications

and are utilised in one or more nations throughout the world. Until 1930, nearly 90% of certified

medications were made from plants5.

During the nineteenth century, chemistry made great progress, and by the turn of the century, the first

synthetic medications had been developed, including phenacetin, urea, barbital, and acetylsalicylic acid

(aspirin). The creation of sulphonamides in the 1930s marked the start of a prosperous phase in

chemotherapy. The age of antibiotics began the next decade, and after WWII, conditions improved for the

development of synthetic chemistry, to the point that medicine was profoundly revolutionised in a few

decades. Since the 1960s, synthetic or fermentation-derived medications have accounted for more than

75% of all conventional medicines, relegating plant-based medicines to a secondary role5.

1.2. Peptic ulcer

Peptic ulcer disease is a prevalent condition around the world6. It is a significant public health issue, both

in terms of morbidity and mortality7. Today, gastric hyperacidity and ulcers are quite widespread, causing

significant human misery. It is caused by an imbalance between corrosive substances in the lumen and

protecting systems in the gastroduodenal mucosa. While it is well established that persistent worry, mental

stress, hemorrhagic surgical shock, burns, and trauma produce significant stomach irritation, the

mechanism is still poorly understood8.

Peptic ulcer is a broad term which includes ulcers of digestive tract in the stomach or the duodenum.

Earlier it was believed that one developed this type of ulcers due to stress and spicy food. However, recent

research has shown that these are just the aggravating factors. The causative agent is infection caused by

the bacteria H. pylori or reaction to certain medicines like non-steroidal anti-inflammatory drugs

(NSAIDs)9. Symptoms of peptic ulcers include abdominal discomfort and pain. Other symptoms include

weight loss, poor appetite, bloating, nausea, and vomiting. Some may also experience blood in stool and

vomit, and black stools that indicate gastrointestinal bleeding10.

The involvement of mucosal factors in peptic ulcers has gotten a lot of attention recently, and the term

"cytoprotection" was coined to describe the physiological mechanisms that protect the stomach mucosa

against acid-pepsin digestion. The majority of these cytoprotective strategies are linked to endogenous

prostaglandin production, at least in part. Inhibition of acid secretion or neutralisation of acid are the two

most common medical treatments for peptic ulcers. Antacids can be used to neutralise stomach acid,

although their efficacy is very temporary. Acid production is effectively inhibited by muscarinic

antagonists such as atropine or pirenzipine. Cimetidine, ranitidine, and famotidine are histamine H2-

receptor antagonists that reduce acid production by 70-80%. Because of the complexity of known
receptors on parietal cells and a variety of second messenger signalling systems coupled to these receptors,

such as adenylate cyclase with histamine receptors and intracellular Ca2+ with acetylcholine receptors,

complete inhibition of parietal cells acid secretion by receptor antagonist is difficult. As a result, inhibiting

the enzyme that causes acid release is the most effective and ideal treatment. The H+ ion pump responsible

for acid secretion in the stomach, the gastric H+-K+ ATPase of the parietal cell, has been discovered as a

pharmacological target for the development of drugs to cure ulcers. Drugs like omeprazole, lansoprazole,

and timoprazole have been demonstrated to be beneficial in the treatment of peptic ulcer disease because

they block the H+-K+ ATPase for a long time. However, such drugs permanently inactivate the ATPase,

and restoring acid secretion after such inhibition necessitates the manufacture of a new pump. The

disadvantage of such inhibitors is that acid secretion can only occur when new ion pumps are produced.

This can be avoided by using reversible inhibitors of H+-K+ ATPase, which allow for more control over

the length of acid secretion suppression. Other medicines that increase mucus formation include

prostaglandins, carbenoxoline, and sucralfate. The negative charge given to mucus by its sulphate radicals

has led to the synthesis of novel compounds such as octasulfate (sucralfate), a basic aluminium sucrose

with characteristics comparable to mucus. As a result of their major activities on mucosal defence factor,

sucralfate and other cytoprotective medications are beneficial in the treatment of peptic ulcer disease.

Furthermore, oxygen-derived free radicals have been linked to the pathophysiology of a variety of illnesses

in a variety of animals. All biochemical groups, including nucleic acids, proteins, free amino acids, lipids,

lipoproteins, carbohydrates, and connective tissue macromolecules, are vulnerable to oxygen radical

damage, which can be reversible or irreversible. In the biological system, these species may have an impact

on cell activities such as membrane function, metabolism, and gene expression; the oxidant of substrates is

primarily affected by free radicals such as reactive oxygen species (ROS) and reactive nitrogen species

(RNS); the oxidant of substrates is primarily affected by free radicals such as reactive oxygen species

(ROS) and reactive nitrogen species (RNS) (RNS). Free radicals are organisms that have one or more

unpaired electrons and can live on their own. These are by-products of aerobic cellular metabolism. The

most common cellular free radicals are superoxide (O2-) and hydroxyl (OH-). Although not free radicals,

hydrogen peroxide (H2O2) and peroxynitrite (ONOO-) contribute to the cellular redox state. These

molecules are collectively referred to as reactive oxygen species (ROS) (ROS). Mitochondrial oxidative

metabolism, enzymatic reactions involving mixed-function oxidases, and auto-oxidation of small

molecules are the main sources of ROS.11,12.

Depending on severity, peptic ulcers are also classified as acute and chronic peptic ulcers. Acute ulcers

involve tissues to the depth of the submucosa. They may arise in the form of single or multiple lesions.

They are found in many sites of stomach and in the first few centimeters of duodenum. Chronic ulcers

penetrate through the epithelial and muscle layers of stomach wall and may include the adjacent pancreas

or liver. In majority of cases, they occur singly in the pyloric antrum of the stomach and in duodenum13.

The pathological findings in both are similar and quite diagnostic.

1.3. Anatomy of an ulcer

A self-lubricating mucus layer protects the lining of the stomach and small intestine, and without it,

gastric acid (hydrochloric acid) would not only consume your food, but also your stomach and intestine.

An ulcer is a lesion or sore that develops along the stomach or intestinal wall, corroding muscle and blood

vessels and causing bleeding, as demonstrated by blood in the stool. If this process is not stopped, germs

and partly digested food can escape into the abdominal cavity, producing irritation and severe discomfort.

How can you tell if you have a stomach ulcer? Ulcers, surprisingly, do not usually cause symptoms.

However, the most typical symptom is a searing discomfort between the breastbone and the navel, which

generally occurs after a meal or when the stomach is empty. After consuming a barium cocktail, x-rays of

the upper GI series or of the esophagus, stomach, and duodenum may be used to make an accurate

diagnosis. Karyn Siegel-Maier uses blood and stomach tissue testing to establish whether H. pylori is

present. (http://www.herbalmusings.com/#!herbs-for-ulcers-/cb6b).

1.4. Causes of ulcer

1.4.1. Helicobacter pylori gastritis

Two Australian researchers discovered the bacterium Helicobacter pylori and deciphered its role in

gastritis and peptic ulcer disease, have been awarded this year's Nobel Prize in Physiology or Medicine.

They revealed that gastritis, and ulceration of the stomach or duodenum, were the result of infection with

some curved Gram negative bacilli14,15.

H. pylori comes in a variety of strains, as well as two different phenotypes. Both produce Vac A, a

vacuolating cytotoxin. Type I additionally possesses a cytotoxin-associated gene (cag A) that may be

required for Vac A cytotoxin transcription, function, or excretion. Ulcer development is linked to the type

I phenotype. Because type II organisms lack cag A, they do not create as much inflammation. Each of
these has the potential to harm cells. 16. Only bacteria that express the cag A antigen have been linked to

ulcer illness, according to Covacci et al. The relationship between H. pylori cytotoxin expression and ulcer

illness is explained by these findings.17,18.

1.4.2. Acid-pepsin secretion

It has been reported that about 50% of gastric ulcer patients are pepsin and acid hypersecretors19. It is the

first line of mucosal defense to prevent bacterial colonization and reduced their ability to entrance in the

mucosal layer20.

1.4.3. Mucus secretions

Mucus secretion is a crucial factor in the protection of gastric mucosa from the gastric lesions and has

been regarded as an important defensive factor in the gastric mucus barrier. A decrease in the synthesis of

sulphated mucus glycoprotein has been implicated in the aetiology of gastric ulcer21.

1.4.4. Gastritis

Gastritis is a condition in which the stomach lining becomes inflamed (irritated). Many things can cause

this, including infection, alcohol, some drugs, and certain allergy and immunological diseases. Gastritis

can be acute (with severe bouts lasting a day or two) or chronic (with symptoms lasting more than a day

or two) (with long-term appetite loss or nausea). In many situations, gastritis goes unnoticed

(asymptomatic). Chronic atrophic gastritis, for example, has been linked to an increased risk of stomach

cancer. Avoiding recognised irritants and using medicine to lower the quantity of stomach secretions are

two treatment options. (www.betterhealth.vic.gov.au).

1.4.5. Local irritants

Smoking, alcohol intake, coffee consumption, and familial occurrences of peptic ulcers in people with

gastric or duodenal ulcers are only a few of the additional variables that cause this condition. According

to epidemiological research, smokers are nearly twice as likely as non-smokers to acquire peptic ulcer

disease. Smoking raises stomach acid secretion and causes duodenogastric reflux, while lowering

gastroduodenal prostaglandin and pancreatic duodenal bicarbonate production.22.

1.4.6. Dietary factors

In experimental models23, different kinds of diet trigger mucosal defence mechanisms. Since the turn of

the century, the usage of dietary essential fatty acids has increased, reducing the incidence of peptic ulcer

disease24. Fresh rice oil protects against stomach ulceration in animal tests, but stored oil is ulcerogenic25.

Intake and handling of rice in diverse parts of the world may also explain peptic ulcers. Salt raises the risk

of death from gastric ulcers but not from duodenal ulcers26. In a Swedish-Norwegian investigation,

duodenal ulcers relapsed more quickly on a low-fiber diet than on a bran-supplemented diet27, dietary

fibre was found to be protective. Milk, on the other hand, appears to have a negative impact on the pace

of duodenal ulcer healing28.

1.4.7. Psychological factors

Peptic ulcer illness is considered a stress-related psychosomatic condition. Stress-induced emotional

problems have long been implicated in the pathophysiology of this disease29. Psychological stress appears

to cause many ulcers and hinder therapy response. This stress likely works as a cofactor with H. pylori. It

may work via increasing stomach acid production or by encouraging unhealthy behavior30.

1.4.8. Endogenous mediators

Several endogenous mediators or chemicals have been found and claimed to be involved in the production

of gastrointestinal lesions, including lipid metabolites, neuropeptides, biogenic amines, reactive oxygen

species, and free radicals.31.

1.4.8.1. Platelet-Activating Factor (PAF)

One of the most effective ulcerogens is PAF32. The sequestration of nuetrophil aggregates in the stomach,

vasoconstriction, production of free radicals, and release of lysosomal enzymes are all implicated in PAF-

induced ulceration33,34.

1.4.8.2 Thromboxane A2 (TXA2) and Leukotriens (LTC4/D4) are produced from arachidonic acid by the

enzymes cyclooxygenase and lipooxygenase. TXA2 mediated stomach mucosal ulceration may be caused

by vasoconstriction, which predisposes the mucosa to disruption by local irritants. Leukotrienes cause

tissue necrosis in the stomach by causing vasoconstriction in the vascular bed in the rat submucosa35.

Pepsinogen secretion from the gastric main cell36 is also stimulated by leukotrienes. It has been shown that
leukotrienes are involved in stress-induced37 and ethanol-induced38 ulcers, which are caused by a

reduction in blood flow and an increase in reactive oxygen species. 39, 1.4.8.3 Histamine 40.

The oxyntic mucosa of humans and other animals contains substantial amounts of histamine, around 40

micrograms per wet weight41. The stomach wall contains histamine, which is a potent stimulator of

gastric secretion42. Excessive histamine release by histamine release or injection of aqueous solution of

histamine, on the other hand, causes gastric and duodenal ulcer43. Because inhibiting histamine receptors

inhibits reserpine, steroid, and NSAID-induced stomach ulcer in humans and experimental animals44,

histamine is also implicated in the other form of ulceration. Psychological stress-induced stomach ulcers

has been observed to be prevented by histamine blockers like Ranitidine. 45,46. \s1.4.8.4 Serotonin is a

neurotransmitter that regulates mood (5-hydroxytryptamine, 5HT)

The gastrointestinal system contains more than 90% of endogenous 5-hydroxytriptamine. In the gut, 5-HT

is stored in endocrine cells and enteric neurons. 5-HT has been proven to have ulcer-producing effects.

Serotonin has been implicated in the ulceration caused by ethanol and reserpine. These ulcerogens

produce a decrease in gastric blood flow and mucus depletion by releasing 5-HT from the stomach

mucosa. 47,48. \s1.4.8.5 Radicals who oppose the status quo

Free radicals are chemical entities with unpaired electrons in their outer orbit that are highly reactive in

nature. When free radicals react with a nonradical, they must generate another free radical. This

consequence is constantly reflected in cells, whether during phagocytosis or in pathological situations. In

aerobic cells, oxygen and its radical derivatives O-2 and OH, H2O2, as well as transition metals (Fe2+,

Cu+) are the most significant reactants in free radical biochemistry. 49. Superoxide radicals are formed

when oxygen is reduced by the transfer of a single electron.

O2 + e – → O2 –

Superoxide radicals can combine with nitric oxide to produce peroxynitrite

O2 – + NO– → ONOO –

H2O2 is produced in biological systems by the dismutation of superoxide after a two-electron reduction of

oxygen.
H2O2 is not a free radical, but it does belong to the category of reactive oxygen species (ROS), which

includes not only oxygen free radicals but also non-radical oxygen derivatives that contribute to the

generation of oxygen radicals. H2O2 is a key chemical in free radical biology because it is quickly broken

down, especially in the presence of transition metal ions, to create the most reactive and destructive

oxygen-free radical, the hydroxyl radical OH.

H2O2 + Fe2+ –→ OH + OH– + Fe3+

The Fenton reaction is an ion catalyzed (Fe2+) process. The Haber-Weiss reaction is a non-catalyzed

process in which superoxide reacts directly with H2O2.

O2 – + H2O2 –→ OH + OH- + O2

Because of the low steady state and concentration of the reactants in biological systems, spontaneous

reactions are less common. Superoxide can be produced via the auto-oxidation of a reduced transition

metal.

Fe2+ + O2 –→ Fe3+ + O –

Cu+ + O2 –→ Cu2+ + O2 –

The hydroxyl radicals can damage components from all biological groups, including nucleic acid, protein,

and free amino acids, lipids, lipoprotein, carbohydrates, and connective tissue macromolecules, reversibly

or permanently. These species might affect cell functions like membrane function, metabolism, and gene

expression. The function of oxygen-derived free radicals in acute and chronic ulceration has been

demonstrated50. Different models of gastrointestinal mucosal injury, such as colitis, ischemia11,

reperfusion stress, and ethanol induced ischemia/reperfusion, have been implicated in neutrophil

involvement in ulcer. In neutropenic animals, ethanol induced injury to the gastric and intestinal mucosa is

significantly reduced. It is also commonly acknowledged that oxygen-derived free radicals cause lipid

peroxidation and cellular membrane damage, as well as the release of intracellular components such as

lysosomal enzymes, which causes additional tissue damage.

Another theory is that free radicals, particularly OH•, cause hyaluronic acid, the main component of the

epithelial basement membrane, to degrade, resulting in mucosal injury. The body has a system in place to

prevent and counteract free radical damage. Superoxide dismutase, glutathione peroxidase, glucose
oxidase, and catalase are examples of natural antioxidant enzymes that perform this. They aid in the

maintenance of the equilibrium between the production of reactive oxygen species and their elimination.

1.5. Ulcer Treatment

1.5.1 Antacids are a kind of antacid.

Antacids are increasingly frequently used for self-medication and are mostly given for symptomatic relief.

They are used to alleviate the discomfort associated with hyperchlorhydria in the stomach. The majority

of antacids include a mix of calcium, aluminium, and magnesium, which all have negative side effects.

Antacids are unsuccessful at neutralising acid in the stomach at low dosages; exceptionally high doses of

antacids are necessary to completely neutralise the excess acid in the stomach. Long-term usage of these

antacids, on the other hand, has been demonstrated to provide considerable mucosal protection51.

Antacids are combined to provide both rapid and prolonged activity, to reduce unpleasant side effects by

using lower doses of each component, and to employ one component to counteract the actions of another

(e.g. laxation versus constipation). Al (OH)3 and Mg (OH)2 is the most prevalent combination.

1.5.2 Histamine antagonist of the H2-receptor

Black and his colleagues made a big breakthrough in the treatment of peptic ulcers with the development

of H2-receptor antagonist in the mid-1972s. H2-receptor antagonists can compete with histamine at all

H2-receptors, although their primary therapeutic application is as a stomach acid secretion inhibitor52.

Stomach volume and pepsin concentration in the gastric content are likewise reduced by H2-receptor

antagonists. In a dose-dependent and competitive manner, an H2-receptor antagonist decreases stomach

acid production evoked by histamine or gastrin. Both basal and food-stimulated acid secretion is reduced

by 90% or more with these medicines. Intrinsic factor secretion from parietal cells is similarly reduced by

H2-receptor antagonists. In treatment, newer H2-receptor antagonists including nizatidine and

famotidine53 are available. Recently, it was discovered that roxatidine, also known as loxatidine, has no

known side effects and is more effective and long-acting.

1.6.3 Muscarinic antagonists .

Gastric acid secretion is reduced by 40-50 percent when using muscarinic antagonists, although stimulated

secretion is suppressed to a lower level. The increased release of histamine and stomach acid caused by

vagal stimulation can be inhibited by nicotine or a muscarinic antagonist (pirenzepine)54. Furthermore, in

the presence of an H2-antagonist, cholinergic agonists can stimulate acid secretion via interacting with the

muscarinic receptor on parietal cells. M1 receptor antagonists are equally effective as atropine or other

nonselective muscarinic antagonists, but they are less prone to cause the side effects associated with

cholinergic blockade (e.g. dry mouth, tachycardia). Pirenzepine and telenzepine are two such medicines

now in clinical trials in the United States.

1.6.4 Proton pump inhibitors.

As long as the cell is not secreting acid, the H+K +ATPase enzyme (proton pump) present in the smooth

membrane structure of the parietal cell termed tubulovesicles is the ultimate mediator of acid secretion.

Acid secretion moves from the cell's secretory canaliculus to the microvilli of the secretary canaliculus. The

shift in c-AMP or (Ca2+) levels in the cell causes this metamorphosis. H+K +ATPase is made up of two

subunits: a and b. The a-subunit is a catalytic subunit that contains 1034 amino acids and has a molecular

mass of 114 KDa. The phosphorylation site, ATP binding site, and proton pump inhibitor binding site are

all found in subunit 'a.' The b-subunit is a glycoprotein with a molecular mass of 60-80 KDa and 291

aminoacids. One of the functions of the b-subunit is to keep the a-subunit in place in the membrane55.

Omeprazole has also been shown to have a clear link between acid secretion suppression and

H+K+ATPase blockade. Inhibition's mechanism has also been thoroughly researched. The inhibition is

caused by the active chemical establishing an irreversible disulfide bond with the SH group of H+K+

ATPase56. Other substituted benzimidazoles, such as timoprazole and picoprazole, decrease acid secretion

in the same way that omeprazole does. Their effectiveness and mechanisms of action have been

investigated, however, omeprazole was shown to be more effective than the other two substances.

Headache, diarrhea, skin rash, and reversible abnormalities in biochemical liver function tests have all

been reported as adverse effects.

The H+K+ATPase enzyme is completely inhibited after long-term usage of omeprazole. When taken

long-term, omeprazole causes gastric cancer, most likely because of total acid suppression, which boosts

gastrin synthesis and causes hyperplasia57. Omeprazole medication in patients leads to achlorhydria

(abnormal acid secretion) and hypergastrenemia (58.1.7). Plants with anti-ulcer action
An ulcer is the most frequent illness on the planet. Even though ulcer is one of humanity's oldest

recognized illnesses and affects a considerable portion of the global population, no significant progress has

been achieved in finding a lasting treatment. The hunt for anti-ulcer activity screening and medication

development has been a never-ending dilemma. Since there have been several reports of plants having anti-

ulcer activity, there is a lot of promise for identifying active anti-ulcer compounds from indigenous plants.

Table 1 lists the reviews on plants with anti-ulcer properties.

Table 1: Indian medicinal plants reported for their ulcer protective activity
 Experimental
S.No Plant Name (Family) Part used Extract Reference
model

Hypothermic
Aegle marmelos restraint stress, Dhuley et al.,
1 Unripe fruit
(Rutaceae) absolute ethanol, 200459
and indomethacin

Aspirin, cold-
Govindarajan
Anogeissus latifolia 50% aqueous resistant stress,
2 Bark et al., 2006 (In
(Combretaceae) alcoholic pylorus ligated,
press)60
and ethanol

Azadirachta indica Subapriya et

3 Leaves Ethanolic
(Meliaceae) al., 200561

Cold restraint
Bocopa monniera stress, aspirin, Rao et al.,
4 Whole plant Juice
(Scrophulariaceae) ethanol, pylorus 20008
ligation

Camellia sinensis Saponin Ethanol and Yoshikawa et

5 Seeds
(Theaceae) fraction indomethacin al.,200562

Cold restraint
stress,
Centella asiatica indomethacin, Sairam et al.,
6 Whole plant Juice
(Umbelliferae) ethanol, pylorus 200163
ligation, acetic
acid

Curcuma longa Ethanol and Kim et al.,

7 Pylori-ligation
(Zingiberaceae) ethylacetate 200564

Ethanol, pylorus
8 Desmodium Ethanolic ligation, aspirin, Dharmani et
gangeticum and cold-resistant al., 200565
(Leguminosae) stress

Aly et
Glycyrrhiza glabra (
9 Root Aqueous Indomethacin
Papilionaceae)
al., 200566

Momordica
Helicobacter Grover et al.,
10 charantia
pylori 200467
(Cucurbitaceae)

Cold restraint
Musa sapientum Methanolic Goel et al.,
12 Pulp stress and anti
(Musaceae) extract 200168
H.pylori activity

Plantago major L. Eksp Klin et

15 Leaves
(Plantaginaceae) al., 200569
2. LITERATURE REVIEW

The annual herbaceous plant Sisymbrium officinale (SO) (L.) Scop. (hedge mustard, originally Erysimum

officinale) belongs to the Brassicaceae family and has a single upright or erect-branched at right angles

stem that is stiff and opaque green or purple with scattered trichomes of 1mm.
 Figure 1: Hedge mustard plant and seeds

2.1. Taxonomic classification

Kingdom: Plantae

Subkingdom: Viridiplantae

Superdivision:Embryophyta

Subdivision: Spermatophytina

Division: Tracheophyta

Class: Magnoliopsida

Order: Brassicales

Family: Brassicaceae

Genus: Sisymbrium

Species: Sisymbrium

2.2. Synonyms: Erysimum officinale

2.3. Common name: Common Hedgemustard, English Watercress, Hairypod Hedgemustard

2.4. Description

It has petiolate, rosette-like basal leaves, or deeply split leaves with serrated margins and a considerably

larger rounded terminal lobe that can reach 90 cm in height. It's a native of Europe and North Africa, but

it's now widely distributed over the globe70. SO is a weed that grows in the wasteland, along roadsides,

and in cultivated and disturbed environments. The lower (basal) leaves are up to 10 cm long, pinnatisect,

with 3–5 pairs of serrated lobes and a wide petiolate terminal lobe. The leaves on the middle and top stems

are smaller, have a short petiole, and are less split and alternating.

The stem is tall, upright, wiry, and has a short internode length. It has short hair and is occasionally

glabrous. The little terminal blooms occur in racemes and have four 2–4mm long golden to light yellow

petals in the form of a cross. There are no bracts on the inflorescence. There are four 2mm long free sepals

and four to six stamens, with the inner four being longer than the outer pair. The fruit is a siliqua with an

upright cylinder-conical form and a peduncle near to the cause. A short stout pedicel (stalk) 1–2mm length

is retained near to the stem. It opens along the 3 ribs releasing from each of the 2 valves at maturity, each

containing many seeds measuring 1–1.5 0.6 0.8mm. 71. \s2.5. Traditional usage and medicinal

characteristics

This plant is used to treat respiratory tract infections such as pharyngitis, laryngitis, cough, aphonia,

common cold, sore throat, and asthma72. It also has antibacterial, antimutagenic, and muscle relaxant

effects, albeit these are less well recognized and used. Furthermore, smokers are advised to consume SO

because of its anti-inflammatory and antioxidant properties73. It's worth noting that SO was first used

more than 2000 years ago in the Greek and Roman periods, when this plant was utilized to cure a variety

of ailments, including mastitis, orchitis, and certain cancers (without solid medical support) 74.

2.6. Phytochemistry The biological effects of SO can be traced mostly to volatile molecules, notably

breakdown products of glucosinolates, an important family of distinctive secondary metabolites found in

many Angiosperms, particularly Brassicaceae75,76. The most significant glucosinolates in SO are

glucoputranjivine, glucojiabutina, napoleiferina, sinalbina, and sinigrin77, which are all sulfated

compounds. Chemical analysis of SO molecular composition revealed that its air dried sections contain

mucilages (10.9%), ash (9.2%), ictiols (8.9%), glucosinolates (0.63%), and flavonoids (0.63%). (0.5

percent,). 78 Isopropyl-glucosinolate makes up roughly 65 percent of all glucosinolates. 75

2.7. Pharmacological action reported

1. Abhishek Tripathi et al. (2021) Peptic ulcer is the term which refers to acid peptic injury of the
digestive tract, and it results in mucosal break reaching the submucosa. Leaves of Capparis
zeylanica are used as counterirritant, rubefacient, as a cataplasm in piles, boils and swellings. The
objective of the present study was to evaluate the antiulcer activity of C. zeylanica ethanolic extract
against chemically induced ulcers. The leaves were extracted with ethanol (50%) as solvent using hot
perforation method. The extract was evaluated against acute and chronic ulcer models. Further,
extract was evaluated for gastric autopsy of animals infected with Helicobacter pylori bacteria. The
genes of rats were evaluated by gel electrophoresis method. Morphology of stomach was also studied
after treatment with plant extract.
2. Ousman Ahmed et al, (2022) Gastric ulcer is a major public health problem globally and associated
with severe complications including hemorrhages, perforations, gastrointestinal obstruction,
and malignancy. Urtica simensis is widely used for traditional management of gastric ulcer in different

parts of Ethiopia. The present study was undertaken to evaluate the anti-gastric ulcer activity of
aqueous and 80% methanol extracts of U.simensis in rats.
3. Sisay Zewdu W et al, (2020) In the pylorus ligation-induced ulcer model, pre-treatment with the
crude extract significantly reduced the degree of gastric secretions, pH, total acidity, and ulcerations
in a dose-dependent manner. Gastro protection offered by the R. nepalensis 400 mg/kg test extract
was comparable to that of the standard. Among fractions, the ethyl acetate fraction at 400 mg/kg had
the highest protection of ulcer but the chloroform fraction was ineffective. In the cold restraint stress-
induced ulcer model, R. nepalensis at 200 and 400 mg/kg reduced the lesion index significantly (P<
0.01). With relevant chronic ulcer model treatment, a dose of R. nepalensis at 200 and 400 mg/kg
healed ulcers significantly with a curative ratio of 53.22% and 54.59%, respectively.
4. P. Pandian et al, (2020) There are several factors that may induce ulcers in human beings such as
stress, chronic use of anti-inflammatory drugs, etc. Though in most cases, the etiology of ulcer is

unknown, it has generally accepted that it is the result of an imbalance between aggressive factors and
maintenance of the mucosal integrity through the endogenous defense mechanism. Thus, the search
for a safe anti-ulcer drug that optimizes these properties is continuing, and part of the search is the
evaluation of medicinal plants for gastroprotective properties. The Malvastrum tricuspidatum has
used in folk medicine for the treatment of inflammation and gastrointestinal diseases. In this
study, we assessed for anti-ulcer activities with aqueous extract and in-vitro method as the acid-
neutralizing capacity and H+/K+ – ATPase inhibition activity method and. In acid-neutralizing
 capacity (ANC), the extract significantly reduced ANC to 9.33 at a concentration of 1500 mg as
compared to 15.7 with standard Aluminium hydroxide + Magnesium hydroxide (500mg). While in
H+ /K+ – ATPase inhibition activity, the extract showed maximum percentage inhibition of 62.18%
at the concentration 100µg as compared to 69.56% with standard Omeprazole.
5. Sajina, V et al, (2021) The plant Calycopteris floribunda Lam is widely distributed in Bangladesh and

India. The antiulcer effect of Calycopteris floribunda Lam leaf have been never studied. Hence the
objective of the study is determining this effect from the Calycopteris floribunda Lam leaf extract.
The preliminary phytochemical screening of leaf extracts indicate in presence of flavonoid, alkaloid,
tannins, and glycosides may accounts anti-ulcer potential. The antiulcer effect is screened in leaf
extract of Calycopteris floribunda Lam on cold water immersion, ethanol and immobilization
induced dose dependent ulcer study. The results get from these study have been shown that leaf
extract of Calycopteris floribunda Lam produce antiulcer effect in cold water immersion, ethanol and
immobilization induced ulcer models. In cold water immersion, ethanol and immobilization induced
model, there is reduction in ulcer index, total acidity, total volume of gastric contents, total protein
concentration and higher concentration of glutathione content and pH of gastric secretion they
compared with control treated group.

3. AIM AND OBJECTIVES

The present study aims to carry out phytochemical investigation and evaluation of the anti-ulcer activity

of Hedge mustard seeds

The present work has been undertaken to fulfil the following objectives:

Preliminary phytochemical studies on the selected plants.

Evaluation of acute and sub-chronic toxicity of extract obtained from Hedge mustard seeds.
Evaluation of the antiulcer activity of Hedge mustard seeds against aspirin-induced ulcers.
4.6.1.1. Animal grouping and treatment:

A total of 30 rats were divided into 6 groups (5 animals per group).

Group 1 received an oral dose (2 mL/kg/day) of propylene glycol for 28 consecutive days. Group 2 received

150 mg/kg/day of aspirin suspended in 3 mL of 1% carboxymethylcellulose (CMC) in water for 3 days

during which the rats fasted for induction of ulcer.

Group 3 rats received cimetidine 100 mg/kg suspended in 3 mL of 1% CMC in water orally for 28

consecutive days followed by 150 mg/kg aspirin suspended in 3 mL of 1% CMC in water for 3 days.

Groups 4, 5, and 6 rats received Hedge mustard seeds extration respectively for 28 consecutive days in

water for 3 days.

Bibliography

1. Tripathi, A., Singh, S. & Mukerjee, A. Antiulcer activity of ethanolic leaf extract of Capparis
zeylanica against chemically induced ulcers. Futur J Pharm Sci 7, 211 (2021).

2. Ousman Ahmed, Teshome Nedi, Ebrahim M. Yimer, Evaluation of anti-gastric ulcer activity of

aqueous and 80% methanol leaf extracts of Urtica simensis in rats, Metabolism Open, Volume 14,
2022, 100172.

3. Sisay Zewdu W, Jemere Aragaw T. Evaluation of the Anti-Ulcer Activity of Hydromethanolic Crude
Extract and Solvent Fractions of the Root of Rumex nepalensis in Rats. J Exp Pharmacol.

2020;12:325-337
4. Yadav A, Mohite S. Screening of In-vitro anti-inflammatory and antibacterial assay of Malvastrum

Coromandelianum. International Journal of Pharma Sciences and Research. 2020;11(4):68-70.

5. Sajina V. Evaluation of Anti-Ulcer Activity of Calycopteris Floribunda Lam Leaf Extract in Wistar

Rat (Doctoral dissertation, RVS College of Pharmaceutical Sciences, Coimbatore).