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 Indicumplant 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
Scoring of ulcers.
|
Serial Number |
Stomach Colour |
Ulcer Score |
|
1 |
Normal colour |
0 |
|
2 |
Red colour |
0.5 |
|
3 |
Red spots |
1 |
|
4 |
Hemorrhagic streaks 1.5 |
1.5 |
|
5 |
3>5 ulcers 2 |
2 |
|
6 |
<5 ulcers 3 |
3 |
Calculationof ulcer Index:
U1 = UN + US + UP x 10-1
U1 = Ulcer Index
UN = Average of number of ulcer per animal
US = Average of severity score
UP = Percentage of animal with ulcer
DETERMINATION OF ACIDITY:
Acidity = n Χ 0.01 Χ 36.45 Χ 1000
Where,
n is volume of NaOH consumed,
36.45 is molecular weight of NaOH,
0.01 is normality of NaOH,
1000 is the factor (to be represented in liter).
Percentage inhibition of ulceration was calculated as below:
% Inhibition of Ulceration= (U.I Control – U.I Test) Χ 100 / U.I Control
Pyloric ligation – 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): Diclofenac sodium x 6
Group – II (Standard): Fomatidine x 6
Group – III: Extract (250mg/kg) x 6
Group – IV: Extract (500mg/kg) x 6
Animal are divided in to four groups, each contain 6 animals Group I having pyloric ligation and receivedDiclofenac sodium (20mg/kg) orally. Groups II received
Fomatidine (10mg/kg) as reference drug for ulcer protective study. Group III and IV received aqueous extract of Xanthium Indicum in a dose of250 and 500mg/kg. After 45 min of aqueous extract of Xanthium Indicum and Fomatidine treatment, pyloric ligation was be done by ligation the pyloric end of stomach of rats of respective groups under ether anesthesia. Animal were allowed to recover and stabilize in individual case and were deprived of water during post operative surgery. After 4 h. of Surgery, rats were sacrificed and ulcer score was done. Gastric juice was collected and gastric secretions studied were performed.
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 Topfer’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 wasNoted. 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
Scoring of ulcer
|
Serial Number |
Stomach Colour |
Ulcer Score |
|
1 |
Normal colour |
0 |
|
2 |
Red colour |
0.5 |
|
3 |
Red spots |
1 |
|
4 |
Hemorrhagic streaks 1.5 |
1.5 |
|
5 |
3>5 ulcers 2 |
2 |
|
6 |
<5 ulcers 3 |
3 |
DETERMINATION OF ACIDITY
Acidity = n Χ 0.01 Χ 36.45 Χ 1000
Where,
n is volume of NaOH consumed,
36.45 is molecular weight of NaOH,
0.01 is normality of NaOH,
1000 is the factor (to be represented in liter).
Percentage inhibition of ulceration was calculated as below:
% Inhibition of Ulceration= (U.I Control – U.I Test) Χ 100 / U.I Control
Statisticanalysis:
The values are represented as the mean±SEM for each group. Statistical signification between treated and control groups were analyzed using a One-way analysis of variance (ANOVA) followed by Dennett’s t-test. Results were considered to be statistically significant at P<0.001.
RESULT:
In the present study, aqueous leaf extract of xanthium indicum was evaluated for its anti-ulcer activity against NSAIDs and pylorus ligation induced gastric ulcer model. The results of study are tabulated in Table-I and Table –II.
Table I
Effect of aqueous leaf extract of xanthium indicum on NSAIDs – induced ulcer in rats
|
Group |
Dose mg/kg |
Ulcer index |
%ulcer protection |
|
Control(diclofenac sodium) |
20 |
4.66±0.18 |
|
|
Standard(Ranitidine) |
100 |
1.05±0.11** |
77.4% |
|
Test-1(Xanthium indicum) |
250 |
1.81±0.23* |
61.15% |
|
Test-2(Xanthium indicum) |
500 |
1.66±0.48** |
64.2% |
Values are mean ± S.E.M. (n = 6), Significant as compared to control P* < 0.01
Table II
Effect of aqueous leaf extract of Xanthium Indicum on Pylorus ligation – induced ulcer in rats
|
Group |
Dose mg/kg |
Ulcer score |
Gastric volume |
Gastric Ph |
Free acidity |
Total acidity |
Ulcer Index |
% Ulcer Protectin |
|
Control(diclofenac sodium) |
20 |
2 |
5.2 |
2 |
110 |
180 |
8.83±0.11 |
|
|
Standard(Fomatidine) |
10 |
0.5 |
6 |
6.7 |
9 |
42 |
1.76 |
80.68% |
|
Test-1(Xanthium Indicum) |
250 |
1.5 |
7 |
6.2 |
48 |
98 |
3.78 |
57.95% |
|
Test-2(Xanthium indicum) |
500 |
0.5 |
6.2 |
7 |
22 |
40 |
1.91 |
78.40% |
P** < 0.05 when compared to control group n=6
In NSAIDs induce ulcer model the plant extract at dose of 250 and 500 mg/kg showed significant gastro protective activity 61.15% and 64.2%compared with standard drug Ranitidine showed 77.4% (Table-I).In ligation induced gastric ulcer Both doses of Xanthium I ndicum extract showed significant reduction in ulcer index, free acidity, total acidity and gastric volume but raised PH of gastric Juice as compared to the control groups. It was showing protection index 57.95% and 78.40% at a dose of 250 and 500 mg/kg respectively compared with standard drugs Fomatidine showed 80.68% (Table-II). To conclude, the aqueous extract of Xanthium Indicum possesses anti-ulcer activity. The aqueous extract of Xanthium Indicum produced both antisecertory and cytoprotective effect.
Ulcer index of NSAIDS induced ulcers
Control group: Control group receives only Diclofenac sodium (20 mg/kg)
Standard group: Standard group receives Ranitidine (100 mg/kg) +Diclofenac sodium (20 mg/kg)
Test -1: Test -1 group animals receive test formulation of Xanthium Indicun of low dose (250 mg/kg) + Diclofenac sodium (20 mg/kg)
Test -2: Test -2 group animals receive test formulation of Xanthium Indicun of higher dose (500 mg/kg) + Diclofenac sodium (20 mg/kg)
Ulcer index of pyloric ligation induced ulcers
Control group: Control group receives only Diclofenac sodium (20 mg/kg)
Standard group: Standard group receives Fomatidine (10 mg/kg) + Diclofenac sodium (20 mg/kg)
Test -1: Test -1 group animals receive test formulation of Xanthium Indicun of low dose (250 mg/kg) +Diclofenac sodium) + Diclofenac sodium (20 mg/kg)
Test -2: Test -2 group animals receive test formulation of Xanthium Indicun of higher dose (500 mg/kg) + Diclofenac sodium (20 mg/kg)
Preliminary Phytochemical Screening:
The MEXI were found to contain alkaloids, carbohydrates, tannins, flavonoids, glycoids, sterols, saponins.
Figure: Effect of Xanthium Indicum on NSAID’S induced ulcer model in rats
A= control (Diclofenac sodium), b= standard (Ranitidine),c=test-1(low dose)
D= test dose-2 (higher dose)
Figure: Effect of Xanthium Indicum on pylorus ligation induced ulcer model in rats
A= control (Diclofenac sodium), b= standard (Fomatidine), c=test-1(low dose)
D= test dose-2 (higher dose)
Conclusion:
In the present study, methanolic extract of leaves of Xanthium Indicum showed a promising antiulcer effect on peptic ulcer models in pyloric ligation and NSAID’S induced methods in rats. Hence oral administration of the methanolic extract of leaves of Xanthium Indicum has the potential to reduce the formation of peptic ulcers.
This property of methanolic extract of leaves of Xanthium Indicum act would be highly beneficial for treatment of various types of peptic ulcers in human beings and reduce the cost burden of the society. However further the active principles of leaves of Xanthium Indicum responsible for this property is too isolated phytochemically and studies with these purified constituents on different ulcer models are waiting to understand the complete mechanism of antiulcer activity.
REFERENCES:
1. P.C.Dandiya, S.K. Kulkarni. Introduction to Pharmacology. Vallab Prakashan New Delhi, 2005. pp. 247
2. Padmaja Udaykumar. Textbook of medical Pharmacology, CBS publishers, New Delhi, 2005 pp. 317.
3. Kirtikar KR, Basu B.D. Indian medicinal plants.2 nd edn, Delhi, Periodical Express Book Agency,1985;Vol 3: 1717-1718.
4. Nadkarni KM, Nadkarnni AK. Indian materia medica.2 nd edn Bombay: Popular Prakashan, 2000; 1: 686-87.
5. Kokate CK. Practical pharmacognosy.4 th edn, New Delhi, Vallabh Prakashan 1994; 4:110-111.
6. S.K.Bhattacharjee.Hand Book of Medicinal Plant. Pointer publishers, Jaipur, 2001, pp. 230.
7. Patil S.S., Bhide A.A. and Gorle A.M.Antiulcer activity and Ant inflammatory Studies on acacia catechu. Indian Drugs 47(2):52-53(2010).
8. Raju. D., Ilango. K., Chitra. V., Ashish. K.Evaluation of Antiulcer activity of Methanolic extract of Terminalia Chebula fruits in experimental rats. Journal of Pharmaceutical Sciences and research1 (3):101-107 (2009).
9. S.K. Kulkarni. Hand book of experimental Pharmacology, Vallabh Prakashan New Delhi, 2002 pp. 149-150
10. H.Gerhard Vogel. Drug Discovery and Evaluation, Springer-Verlag Berlin Heidelberg, New Your 2002 pp. 868.
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