INTRODUCTION:

Gastro duodenal ulcer and Gastric hyperacidity are the burning issues now-a-days. when the balance between gastro protective and gastro aggressive factors are lost then gastric lesions will develop. Few major gastric aggressive factors are pepsin Helico bactor pylori, bile salts and production of acids in the stomach. The pathological condition for this is due to hyper secretion of gastric acid, which mainly occurs due to elevated release of Hydrochloric acid (HCl) from the parietal cells of the gastric mucosa through the proton pump (potassium hydrogen ATPase). The mainly involved defensive factors are the Prostaglandins and secretion of bicarbonates from mucosa. when some gastro protective factors are lost, even the normal rate of acid secretion causes ulceration in the breached mucosa.

The modern approach to control gastric ulceration is to inhibit gastric acid secretion, to promote gastro protection, stimulate epithelial cell proliferation for effective healing and block apoptosis. Anti-ulcer drugs such as histamine H2 -receptor blocker (Ranitidine, Famotidine), Proton pump inhibitors (Omeprazole, Lansoprazole) are used to control gastric acid secretions, and acid related ulcers caused by Stress, NSAID'S, and H.Pylori but there are reports of adverse effects and relapse in the long run¹.

Herb is an innumerable wealth of nature not only from the global environmental perspective but also from the medicinal point of view. It plays a significant role in mitigating the disease resistant ability and conflict against various un favourable metabolic activities within the living system. Herbal medicine is the main source of about 75-80% of the world population, mainly in the developing countries, for primary health care because of, better compatibility better cultural acceptability with the human body and lesser side effects². The active chemical components present in them are a part of the physiological functions of living flora and hence they are believed to have better compatibility with the Human body. Because of all new re-emerging infectious diseases there is a urgent need to discover new compounds with diverse chemical structures and novel mechanisms of action.

Therefore, researchers are increasingly turning their attention to folk medicine, looking for new leads to develop better drugs³. Allium cepa belongs to the Liliaceae family, cultivated from past five thousand years and have been used in herbal medicine and as an indispensable flavoring agent and as a vegetable⁴. The Allium cepa (onion bulb) consists of 2-Methyl-but-2-en-1-al, 28-Iso-fucosterol, Allicin, Pyruvic acid, Raffinose, Rhamnose, Succinic acid, etc. Pharmacological studies of Alliumcepa were been proved that it possess Antituberculosis⁵, Antimicrobial⁶, Anti-diabetic⁷ and Invitro bacterial⁸ activity. It has recently been the source of much interest because of its antithrombotic, hypolipidaemic, hypotensive, diaphoretic, antibiotic, antidiabetic, antiatherogenic, and anticancer medicinal properties. There was so many of the researches have been carried out on the bulbs of Allium cepa but, no works have been done on the aerial leaves of the Allium cepa. Therefore we made an attempt to evaluate gastro protective activity of Allium cepa aerial leaves⁹.

Plant Description:

Allium cepa leaves:

History:

The onion has been cultivated for over five thousand years and has been used in herbal medicine and as an indispensable flavouring agent or as a vegetable that is cooked or eaten raw. The term Onion is derived from the Latin word, unio, which means "one large pearl", and it is interesting to note that the Chinese called the Onion the "jewel among vegetables”¹⁰.

The ancient records are from Egypt, where it was cultivated during the Old Kingdom. Onions appear as carvings on pyramid walls and in tombs from the third and fourth dynasties (2700B.C.) they were usually depicted on offering tables since fourth dynasty. Numerous remains have been found from the era of the Egyptian New Kingdom (since1580 BC). It was being used for funeral offerings and has frequently been found attached to or within mummies. It must have been important in the daily diet of many people¹¹.

In India since 6^th century BC there are reports regarding onion in articles. In the Europian countries, onion was commonly cultivated as a garden plant. Different cultivators of it are mentioned in garden catalogues from the 9th century A.D e.g., during the reign of Charles the great, in the famous "Capitulars de villis". But only during the middle ages it was grown as crop on large scale and gained popularity. It is said to have been introduced in Russia amid 12th and 13th century.

Chemical Constituents:

The leaf on steam distillation yielded an essential oil (0.006%). The oil contains two new cyclic cis- and trans-3, 5-diethyl-1, 2, 4-trithiolanes. The carotene is also found in flowers. The other compounds identified as constituents of leaf are as follows:

Ascorbic acid, Caffeic acid, Citric acid, Ferulic acid, Fructose, Glucose, Malic acid, Methanol, Oxalic acid, Para-coumaric acid, Para-hydroxybenzoic acid, Propionaldehyde, Protocatechuic acid, Raffinose, Sinapic acid, Succinic acid, Sucrose¹².

Experimental Animals:

Adult albino rats of wistar strain of either sex weighing between 150-200g were used for study. The animals were housed in clean, sterile polypropylene cages in a well ventilated room under hygienic conditions and were exposed to 12 hours a day and night cycle. The animals were fed with commercial rat pellet feed and were given water and libitum.

Experimental Design:

20 Adult albino rats were divided into four groups of 5 animals in each group as follows:

Group-I (n=5): Controlled rats received 96% ethanol.

Group-II (n=5): Induced rats received standard drug (Omeprazole).

Group-III (n=5): Ulcer rats received low dose MEAC (100mg/kg).

Group-IV (n=5): Ulcer rats received high dose of MEAC (200mg/kg).

Extraction by Soxhlet apparatus:

Soxhlet extraction through the process of continuous extraction in which the same solvent can be circulated through the extractor for several times. This process involve extraction followed by evaporation of the solvent. The vapours of the solvent are taken to a condenser and the condensed liquid is return to the drug for continuous extraction.¹³

Soxhlet apparatus designed for such continuous extraction which consists of body of extractor attached with a side tube and siphon tube. The extractor from the lower side can be connected to distillation flask and the mouth of the extractor is attached to a condenser by the standard joints. The crude drug is packed in the soxhlet apparatus directly or in a thimble of filter paper or fine muslin. The thimble diameter corresponds to the internal diameter of the soxhlet extractor. Extraction convention is set up by fixing condenser and a distillation flask. Initially for the setting of the powdered drug, solvent is allowed to siphon once before heating. To avoid bumping of the solvent freshly activated porcelain pieces are added to the flask. The vapours pass through the side tube and the vapours are condensed, liquid gradually raises in the extractor and in the siphon tube. A siphon is set up as the liquid reaches the point of return and the contents of the extraction chamber are transferred to the flask. The cycle of solvent evaporation and siphoning back can be continued as many times as possible without charging the solvent, so as to get efficient extraction. So in present study we used methanol as solvent and extracted by using Soxhlet apparatus¹⁴.

ANTIULCER ACTIVITY:

Ethanol induced model and Pylorus ligation method was used for its analysis¹⁵.

ETHANOL INDUCED MODEL:¹⁶

In this method gastric ulcers were induced by Ethanol induced model. Animals were divided into four groups (n= 5). Group I received no treatment, served as control animals. Group II received standard drug Omeprazole (20mg/kg body weight, per oral). Group III received low dose of MEAC (100mg/kg, per oral), Group IV received high dose of MEAC (200mg/kg, per oral). The animals of all groups were starved for 16h with free access to drinking water. Drugs were given half an hour prior to ethanol (1ml/200gm, orally) administration. After 1hr to ethanol administration rats were sacrificed and stomach was removed. It was opened along greater curvature and observed for lesions¹⁷.

PYLORUS LIGATION MODEL:

Gastric ulcers were induced by pylorus ligation. Animals were divided in four groups (n= 5). Group I receives no treatment, serves as control animals. Group II received standard drug Omeprazole (20mg/kg body weight, per oral) one hour prior to pylorus ligation. Group III received low dose of MEAC(100mg/kg, per oral), Group IV received high dose of MEAC(200mg/kg, per oral).Animal were fasted 24 hrs before to the experiment, Pyloric ligation was done by ligating the pyloric end of the stomach of rats 1 h after drug administration. 6 h later the ligated rats were sacrificed by decapitation and the abdomen was opened, content in the stomach was evacuated and centrifuged at 2000rpm, 3 min for assessing parameters like, gastric pH, total acidity and free acidity. The stomach was washed under running tap water and hooked onto a cork plate¹⁸.

The parameters monitored in pylorus ligation model were:

Total acidity, Free acidity Gastric pH, Acid volume,

Total acidity¹⁹:

A known amount of gastric residue was titrated with 0.1 N NaOH. The total acidity however was determined by titration using phenolphthalein (1% alcoholic) as indicator. Reading was taken (ml NaOH) for total acidity

Y = ml of 0.1 N NaOH x 10

Where,

Y = Total acidity (mEq/L)

Free acidity¹⁹:

A known amount of gastric residue was titrated with 0.1 N NaOH. Add two drops of Methyl orange reagent which changes to a salmon colour when all the free hydrochloric acid was neutralized. Reading was taken (ml NaOH) for free acidity

Y = ml of 0.1 N NaOH x 10

Where,

Y = Free acidity (mEq/L).

Calculation²⁰:

Mean ulcer score for each animal was expressed as ulcer index. The percentage of ulcer inhibition was determined as follows:

Control mean ulcer index - Test mean ulcer index Inhibition of ulcer (%) = x100

Control mean ulcer index

Statistical Analysis:

Statistical analysis was carried out using Graph pad prism software. All values were expressed as Mean + Standard mean error. Groups of data were compared with one way analysis of variance followed by Dunnett test. Values were considered statistically significant at p<0.01²¹.

RESULT AND DISCUSSION:

Results of preliminary phytochemical Investigation

Table: 1: GLYCOSIDE TEST

S. No	Test	*Methanolic extract of Allium cepa* leaves**	*Aqueous extract of allium cepa* leaves**
1	Balget’s test	Positive	Positive
2	Legal’s test	Positive	Positive
3	Killar-killani test	Positive	Positive
4	Raymond’s test	Positive	Positive

Table: 2: ALKALOIDS TEST

S. No	Test	*Methanolic extract of Allium cepa* leaves**	*Aqueous extract of allium cepa* leaves**
1	Mayer’sTest	Positive	Negative
2	Dragendorff’s Test	Negative	Negative
3	Wagner’s Test	Positive	Positive
4	Hager’s Test	Negative	Negative

Table: 3: FLAVANOIDS TEST

S. No	Test	*Methanolic extract of Allium cepa* leaves**	*Aqueous extract of allium cepa* leaves**
1	Shinoda Test	Positive	Negative
2	Lead acetate Test	Negative	Positive
3	Sodium hydroxide Test	Positive	Positive
4	Zn & Hcl Test	Positive	Positive

From the above phytochemical investigation we came to know that there was the presence of flavanoids, alkaloids and glycosides

Table:-4: Extract of Methanolic Extract of Arial Leaves of Allium cepa on Ethanol Induced Gastric Ulcers:

Groups	Treatment	Gastric Index	Protection (%)
I	Control	15.58±0.63	-
II	Standard	2.48±0.22	84.08
III	LDMEAC	3.5±0.13	77.5
IV	HDMEAC	2.8±0.10	82.02

LDMEAC = Low Dose Of MEAC, HDMEAC = High Dose Of MEAC

Values Are mean±Sem For Five Animals In Each Group.

P<0.01concedered Stastically Significant As Compared With Control Group.

Effect of Methanolic Extract of Arial Leaves of Allium cepa on Ethanol Induced Gastric Ulcers :

Table:-5: Effect of Methanolic Extract of Arial Leaves of Allium cepa on Various Parameters in Pyloric Ligation Induced Gastric Ulcers:

Groups	Treatment	Ulcer Index	Protection (%)	pH Of Gastric Juice	Gastric Volume(ml)	Total Acidity(mEq/I)	Free Acidity(mEq/I)
I	Control	8.64±0.19	-	2.56±0.17	1.72±0.086	99.05±1.14	106.5±1.37
II	Standard	3.02±0.07	65.04	6.06±0.05	0.44±0.016	15.92±0.89	24.5±1.05
III	LDMEAC	4.04±0.16	54.2	4.96±0.16	0.81±0.018	26.50±0.89	36.73±1.63
IV	HDMEAC	3.38±0.08	60.8	5.8±0.23	0.62±0.017	19.16±0.18	30.07±0.39

LDMEAC = Low Dose Of MEAC, HDMEAC = High Dose Of MEAC

Values Are mean±Sem For Five Animals In Each Group.

P<0.01concedered Statistically Significant As Compared With Control Group.ss

Effect of Methanolic Extract of Arial Leaves of Allium cepa Various Parameters in Pyloric Ligation Gastric Ulcers :

CONCLUSION:

In this study we observed that, Methanolic extract of leaves of Allium cepa (MEAC) provides significant antiulcer activity against gastric ulcers in rats. Methanolic extract of leaves of Allium cepa showed significant reduction in the ulcer index, when compared to control. The antiulcer activity of extract may be attributed to their flavonoids content and antioxidant property. These findings indicate that MEAC display potential antiulcerogenic activity. Its use in indigenous medicine should be scientifically scrutinized with further research.

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Received on 04.03.2022 Modified on 15.09.2022

Asian J. Research Chem. 2023; 16(2):123-127.

DOI: 10.52711/0974-4150.2023.00020