ANTI ULCER ACTIVITY OF METHANOLIC EXTRACT OF LEAVES OF BOSWELLIA OVALIFOLIOLATA BAL&HENRY

About Authors:
M.Dhanalakshmi*, B. Maddilety
Department of pharmacology, Krishna Teja Pharmacy College,
Chadhalawada Nagar, Renigunta Road, Tirupathi-517 507,
Andhra Pradesh, India.
*Lakshmi.dhana17@gmail.com

ABSTRACT:
Boswellia ovalifoliolata bal&henry is an endemic plant that has been suggested as useful in the treatment of various diseases (anti diabetic, anti inflammatory, antimicrobial , antifungal, antibacterial).In the present study, anti ulcer activity of MELBO was studied by pylorus ligated,aspirin induced gastric ulceration in albino rats. Anti ulcer activity was evaluated by pylorus ligation, aspirin induced ulcer method by using standard ranitidine (27mg/kg).The anti ulcer activity was assessed by determining the ulcer index, pH and acidity. MELBO (400mg/kg) produced a significant reduction in the ulcer index, PH and acidity when compared with ulcer control group and standard group. MELBO reduces ulcer, there by the present study indicates that boswellia ovalifoliolata bal&henry showed protective activity on gastric ulcer.

Reference Id: PHARMATUTOR-ART-1370

INTRODUCTION:
Peptic ulcer disease is occurred due to an imbalance between mucosal defense factors (bicarbonate,mucin,prostaglandin,nitric oxide, other peptides and growth factors) and injurious factors(acid and pepsin)⁽³⁾.

Anti ulcer drugs in the treatment of gastric ulcer is usually over showed by various side effects. Example proton pump inhibitor (omeprazole, lansoprazole) can cause nausea, abdominal pain, constipation and diarrhea. Medicinal plants have the main source of new drug candidates for the treatment of gastric ulcers. The medicines are considered safer because of the natural ingredients with no or less side effects.

Boswellia ovalifoliolata bal&henry is an endemic plant that has been suggested as useful in the treatment of various diseasessuch as antidiabetic⁽⁴⁾, anti-inflammatory, antimicrobial^{(7, 9)}, antifungal⁽⁶⁾, antibacterial⁽⁵⁾.  

Material and Methods:
Plant material:
The leaves of Boswellia ovalifoliolata bal&henry was collected from the hills of tirumala region, chittoor (dist).A.P,India. The plant was identified and auntheticated by Dr.Madhava chetty, Assisatant professor, Departmant of botony, S.V. University, Tirupathi. The leaves of the plant were stored in herbarium at the college for further reference.
Preparation of extract:
The leaves of Boswellia ovalifoliolata bal&henry was collected, washed, cleaned and shade dried. The dried leaves were powdered with the help of mechanical mixer and passed through a 40 –mesh sieve to obtain coarse powder. The weighed quantity of coarsely powdered material was extracted with methanol by using soxhlet apparatus. After completion of extraction, it was undergone distillation under reduced pressure and the remaining solvent was removed by evaporation to dryness on a water bath. Residue was obtained and it was kept in a dessicator and used for further experiment⁽⁸⁾. The percentage yield from 60gm was found to be 2.98%.
Phytochemical screening:
The presence of phytochemical constituents in the MELBO was tested by using the standard methods⁽²⁾.These standard methods revealed the presences of glycosides, flavanoids, steroids, alkaloids.

Evalution of antiulcer activity:
Requirements:
a.      Animal:
Albino rats (150-200g) of either sex were used for the experiment. They were kept in the animal house in a controlled room temperature at 25+2 c, relative humidity 44-56%, light and dark cycles of 10 and 14 hr, respectively for 1 week before the experiment. The animals were grouped and housed in polyacrylic cages for the further experiment.
b.      Drugs and Chemical:
Extract:  The MELBO was dissolved in 1 % tween 80 as a vehicle and administered P.O in a dose of 200mg/kg and 400mg/kg.
Standard drug: The ranitidine was dissolved in 1% tween 80 and administered i.p in a dose of 27mg/kg.
c.       Acute Toxicity studies :
The Acute Toxicity studies were performed in order to establish the therapeutic index of a test drug. The experiment was conducted according to the OECD, 423 guidelines. It was administered as 5,100, 1000 and 2000 mg/kg.
d.   Experimenal design:
Rats were divided in to 4 groups. Each group contains 6 rats and treated with        the following drug for five successive days and 6^th day aspirin was administered and to evaluate the antiulcer activity.
Group 1: Control    - Normal saline (1ml/kg)    
Group 2: Ranitidine - 27mg/kg (standard control)
Group 3: MELBO   -      200mg/kg
Group 4: MELBO   -      400mg/kg.

ASPIRIN INDUCED ULCER METHOD:
Aspirin at a dose of 200mg/kg (20mg/ml suspension in 1%CMC (Carboxy methyl cellulose)) was administered orally to 18hr fasted animals. After 1hr the test drug was administered on 6^th day. The ulcers were scored after 4hrs. The stomach was taken out and cut open along the greater curvature and ulcers were scored by a person in the glandular portion of the stomach. The ulcer index was calculated by adding the total number of ulcers per stomach and the total severity of ulcer per stomach ⁽¹⁾.

PYLORUS LIGATION INDUCED ULCER METHOD:
This is the most acceptale method ⁽¹⁾.Rats were divided in to 4 groups. Each group contains 6 rats. Each weighing about 150-200gm and fasted for 4hr with free access to water. Pylorus ligation induced uler method was performed under diethyl ether anaesthesia to each animal. Animals were given to MELBO extract of 200mg/kg and 400mg/kg b.w. ranitidine was prepared in 1% tween 80 suspension as a vehicle orally immediately after pylorus ligation. Animals were sacrified 6hrs later the stomach was carefully removed and gastric contents were collected. The gastric juice was centrifused at 3000 rpm for 30 mins and then volume of gastric juice was measured. Acidity in the supernant was determined by titration with0.01N NaoH and expressed as m.eq/l. the stomach was cut open along the greature curvature and pinned on a soft board for evaluating gastric ulcers and ulcer index is calculated using formula:
Ulcer index = 10/x
Where x = Total mucosal area / Total ulcerated area.

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RESULTS:
ASPRIN – INDUCED GASTRIC ULCER METHOD:

MELBO: Methanolic extract of Boswellia ovalifoliolata Bal & Henry leaves
Table 1: Values are expressed in Mean + SEM, (n=6), when compared with control,
P<0.05, P<0.01, P<0.001 *, **, *** respectively, Oneway ANOVA followed by Dunnet’s t – Test.

1. Pylorus Ligated induced ulcer Method :

MELBO: Methanolic extract of Boswellia ovalifoliolata Bal & Henry leaves

Table 2: Values are expressed in Mean + SEM, (n=6), when compared with control, P<0.05, P<0.01, P<0.001, *, **, *** respectively, Oneway ANOVA followed by Dunnet’s t – Test

N= 6 animals in each group.

MELBO: Methanolic extract of Boswellia ovalifoliolata Bal & Henry leaves

Table 3: Values are expressed in Mean + SEM, (n=6), when compared with control,P<0.05, P<0.01, P<0.001 *, **, *** respectively, Oneway ANOVA followed by Dunnet’s t – Test

DISCUSSIONS:
The anti ulcer activity of the plant of methanolic extract of leaves ofBoswellia ovalifoliolata bal&henry was evaluated by employing pyloric ligation induced ulcer method & aspirin induced ulcer methods. Ulcers results from extrinsic factors like Helicobacter pylori, Non steroidal anti-inflammatory drugs (NSAIDs), Tobacco smoking, Alcohol, Stress and Intrinsic factors like Acid & Pepsin, Acid Reflux. These factors can be used to induce ulcer in animal models.

The general ulcer inducing methods are pyrolic ligation method, aspirin induced ulcer, cold restraint stress induced ulcers, ethanol induced ulcers, cold water immersion method, swimming stress ulcer method. These models represent some of the most common causes of gastric ulcer in humans.

From the above results, we observed that methanolic extract of leaves of Boswellia ovalifoliolata bal&henry provide significant anti ulcer activity by pyloric ligation induced method & aspirin induced methods against gastric ulcers in rats.

conclusion:
In the present  study, methanolic extract of leaves of Boswellia ovalifoliolata bal&henry showed  a promising antiulcer effect on peptic ulcer models in pyloric ligation and aspirin induced methods in  rats. Hence oral administration of the methanolic extract of leaves of   Boswellia ovalifoliolata bal&henry have the potential to reduce the formation of peptic ulcers.

This property of methanolic extract of leaves of Boswellia ovalifoliolata bal&henry act would be highly beneficial for treatment of various types of peptic ulcers in human beings and reduce the cost burden of the society. How ever Further the active principles of Boswellia ovalifoliolata bal&henry leaves 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:
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5. N.Savitramma, M. Linga Rao and P. Suvarnalatha Devi, Evaluation of Antibacterial Efficacy of Biologically Synthesized Silver Nanoparticles using Stem Barks of Boswellia ovalifoliolata Bal. and Henry and Shorea tumbuggaia Roxb. Journal of Biological Sciences 2011, 11: 39-45.
6. N.Savitramma, Antifungal efficacy of silver nanoparticles synthesized from the medicinal Plants), 2011.
7. N. Savithramma1M. Linga Rao*1, K. Rukmini2 and P. Suvarnalatha devi, Antimicrobial activity of Silver Nanoparticles synthesized by using Medicinal Plants,   International Journal of ChemTech Research CODEN( USA): IJCRGG ISSN : 0974-4290 Vol. 3, No.3, pp 1394-1402, July-Sept 2011.
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