EVALUATON OF ANTI ULCER ACTIVITY OF XANTHIUM INDICUM BY USING EXPERIMENTAL RATS

 

ABOUT AUTHORS:
P.Monica1*, DR.Nikunjal Mishra2
1Yalamarthy college of pharmacy, Visakhapatnam, A.P
2HOD. Department of pharmacology Yalamarthy College of pharmacy, Visakhapatnam, A.P
*monica_reddy2308@yahoo.com

ABSTRACT:
The cause of ulceration in patients is mainly due to hyper secretion of gastric juice and also due to hyper secretion of pepsin. In traditional system of medicine a number of herbal preparations have been used in the treatment of peptic ulcers. Xanthium Indicum have been extracted by methanol and evaluated for the antiulcer activity of NSAID’s induced and pylorus ligation of rats. The extract reduces ulcerative lesions, gastric volume, free and total acidity as well as gastric ulcers in the NSAID’s induced and pylorus ligated rats and the effects were compared with standard drug. The extract of xanthium indicum at a dose of 250mg/kg and 500 mg/kg produced significant inhibition of gastric lesion induced by NSAID’s induced and pylorus ligation in rats.

Reference Id: PHARMATUTOR-ART-1986

INTRODUCTION:
A localized loss of gastric as well as duodenal mucosa leads to the formation of peptic ulcers. It arises when the normal mucosal defensive factors such as mucus, mucosal blood flow, formation of HCO3 and PGE2 are impaired. Also by the aggressive factors includes acid, pepsin, NSAID’s and helicobacter pylori. A number of drugs are available for the treatment of peptic ulcer but its clinical evaluation shows the evidence of relapses side effects and drug interactions. This has been the rational for the development of new antiulcer drugs and search of novel molecules. Peptic ulcers are a lesion of the gastric/duodenal mucosa occurs at a site where mucosal epithelium is exposed to acid and pepsin.Peptic ulcers occurs due to imbalance between the offensive (gastric acid secretion) and defensive (gastric mucosal integrity) factors .Factors such as stress, smoking, nutritional deficiency and ingestion of NSAID’s all can increase the incidence of gastric ulcers or due to pathological condition such as Zollinger Ellison Syndrome. It is also reported that prolonged anxiety, emotional stress, hemorrhagic surgical shock, burns and trauma are known to cause severe gastric irritation. Xanthium Indicum commonly known as Marulamathangi in Telugu belongs to family composite ,small shrub 25-30cms found in waste lands and road sides it shows flowers and fruits throughout the year. leaf  has been found to show, alterative, anti synthalitic, astringent, diuretic, ulcer and seed has found to show cooling, small pox. The extract of this plant reduces ulcerative lesions, gastric volume, free and total acidity.

A peptic ulcer is an open crater or sore that develop in the inner lining (mucosa) of the stomach or the duodenum. A coating of mucus and other chemicals normally shields the stomach and duodenum from digesting themselves. When these protective mechanisms are disrupted, powerful digestive acids can erode into the lining of these organs and cause peptic ulcer.

Peptic ulcer is a major gastro-intestinal disorder caused by an imbalance between offensive (gastric acid, pepsinogen secretion) and defensive (mucus secretion, gastric mucosal integrity) factors. It is a round or oval scor where the lining of the stomach or duodenum has been eaten away by stomach acid and digestive juices.

A peptic ulcer, also known as peptic ulcer disease is an ulcer of an area of the gastrointestinal tract that is usually acidic and thus extremely painful. If it is in stomach, it is referred to as a gastric ulcer. If it is in duodenum, it is called a duodenal ulcer.

Duodenal ulcers are more common than gastric ulcer and usually occur in people under 50.Gastric ulcers are more common in people aged over 50.

5-10% of people worldwide suffer from peptic ulcer at least once in their lifetime.

Peptic ulcer affects both men and women.

MATERIALS:

PLANT MATERIAL:
Xanthium Indicum
plant was collected in the month in the month of  March, 2013  it was commonly found in waste lands and road sides and was authenticated by Dr.k. Madhava Chetty assistant professor, department of botany ,sri venkateswara university,Tirupati .It was collected from the areas of Rangampeta, Tanapalli, Tirupathi throughout the year. Fresh plant material was collected on a filter paper sheets and dried under shade at room temperature until with the changing of color of filter paper and then milled to a coarse powder by mechanical grinder.

Where a voucher specimen has been preserved for further identification

EXTRACT PREPARATION:
The finely grounded powder of Xanthium Indicum was packed into sox hlet column and extracted with suitable quantity of methanol for 48 hours i.e. 2 days by maintaining suitable temperature conditions to obtain methonolic extract of Xanthium Indicum

After this extract was filtered through a what man filter paper no.1 and concentrated under reduced pressure at a low temperature just prior to use the substance was dissolved in dimethyl sulphoxide. Extract so obtained stored in a air tight container in refrigerator for further experimental studies.

PRELIMINARY PHYTOCHEMICAL SCREENING:
Methonolic extract of Xanthium Indicum were subjected to preliminary phytochemical screening for the detection of various plant constituents

EXPERIMENTAL ANIMALS:
Number of adult healthy albino Wister rats of weighing between 160-250gms were used for the study .The animals were housed in standard condition (temperature 24+/-2 with 50-60%retative humidity and a 12 hours light dark cycle).The entire animals had free access to water and normal pelleted diet (Hindustan lever ).The study was approved by Institutional Animal Ethical Committee (IAEC) and was in accordance with the guideline of Committee for the Purpose of Control and Supervision of Experimental Animal.

CHEMICALS:
All the drugs and chemicals used in this experiment should be of analytical grade only

Ranitidine (Research lab grade)

Methanol (Research lab grade)

Di ethyl ether (Fine chem. Industries, Chennai)

Diclofenac sodium (research lab scale)

Fomatidine (Research lab scale)

ACUTE TOXICITY STUDIES:
Acute toxicity studies were performed according to organization for economic co-operation and development guide lines. Animals were divided into groups (n=).The animals were fasted for 4hrs with free access to water only .The extract was administered orally in doses of 250mg/kg and 500 mg/kg to different groups of mice and observed for 14 days for mortality for 24 hrs and physical/behavioral changes for 12 hours .No mortality was found even at   250 mg/kg dose .The dose of that   250 mg/kg was selected for that activity.

EVALUATION OF ANTI-ULCER ACTIVITY:
Two animal models (NSAID’s induced and pylorus ligation) were employed to evaluate the anti ulcer activity of Xanthium Indicum leaf extract.

NSAIDs – induced ulcer:
Healthy female Wister albino rats of weighting between 160-200gm were taken for the studies. The animal were divided in to four groups (each contain 6 animal) us follow

Group – I (Control): Distilled water x 6

Group – II (Standard): Ranitidine x 6

Group – III: Extract (250mg/kg) x 6

Group – IV: Extract (500mg/kg) x 6

The animal in all the groups were kept for 24 h.fasting after that animal of all groups’ received diclofenac sodium (NSAIDs, 20mg/kg). The oral feeding of water and diclofenac sodium was continued for 3 days, the animal of II, III and IV were administered with ranitidine (100mg/kg),Xanthium Indicum extract (250mg/kg), Xanthium Indicum extract ( 500mg/kg )respectively after 3 hr. of diclofenac sodium. On 4th day the animals’ weresacrificed, stomach were removed and cut along the greater curvature to measure the ulcer index.

Methods for biochemical estimation of gastric juice

Collection of gastric juice:
Gastric content collected from NSAIDS induced rats was collected and the volume of gastric juice as well as pH of gastric juice was noted. The gastric juice was subjected to biochemical estimations as follows:

Determination of pH
An aliquot of 1 ml gastric juice was diluted with 1 ml of distilled water and pH of the solution was measured using pH meter

Determination of free and total acidity
1 ml of gastric juice was pipette into a 100 ml conical flask, 2 or 3 drops of Toper’s reagent was added and titrated with 0.01N Sodium hydroxide until all traces of red color disappears and the color of the solution turns to yellowish orange. The volume of alkali added was

Noted. This volume corresponds to free acidity. Then 2 or 3 drops of phenolphthalein solution was added and titration was continued until a definite red tinge appears. Again the total volume of alkali added was noted now this volume corresponds to total acidity.

Macroscopic evaluation of stomach: The stomachs were opened along the greater curvature, rinsed with saline to remove gastric contents and blood clots and examined by a Χ5 magnifier lens to assess the formation of ulcers. The numbers of ulcers were counted. Ulcer scoring was undertaken according to Vogel

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