anti-ulcerogenic potential of natural and synthetic

www.wjpps.com Vol 5, Issue 12, 2016.

905

Kabilan et al. World Journal of Pharmacy and Pharmaceutical Sciences

ANTI-ULCEROGENIC POTENTIAL OF NATURAL AND SYNTHETIC

POORA PARPAM AGAINST INDOMETHACIN INDUCED GASTRIC

ULCER IN WISTAR RATS

N. Kabilan*1 and M. Murugesan

2

*1Department of Siddha, The Tamil Nadu Dr.M.G.R. Medical University, Chennai, Tamil Nadu,

India.

2Former Dean, National Institute of Siddha, Chennai-600047

ABSTRACT

Peptic ulcer disease is a term used for both stomach and duodenal

ulcer. In clinical practice, peptic ulcer disease is most commonly

known as a gastrointestinal disease which is interlinked with the acute

and chronic inflammation of the gastric and duodenal epithelium.

According to an estimation of the WHO, almost 80% of people

globally are treated by traditional medicine. Siddha system of medicine

is considered to be one of the oldest rejuvenating therapies known to

mankind since centuries back. Siddha formulations have consistently

proven its efficacy particularly in the management of gastric ulcer

disease. In the present study, anti-ulcer property of siddha formulations

natural and synthetic Poora parpam was evaluated against

Indomethacin induced gastric ulcer model in rats. The result obtained from the study clearly

shows that treatment with Natural and Synthetic Poora parpam significantly reduced the

ulcer lesion index at dose level of dose of 1.15 and 2.30mg / kg. Similarly rats treated with

standard drug Ranitidine (100 mg / kg) showed higher level of reduction in ulcer index when

compare to positive control group. Standard drug Ranitidine (100 mg / kg) treated group has

shown the highest percentage 69.9 % inhibition of ulcer ,where as Natural Poora parpam

exhibit 38.43 and 57.99% inhibition at the dose of 1.15 and 2.30 mg / kg respectively.

Similarly rats treated with Synthetic Poora parpam exhibit 29.12 and 48.21% inhibition at

the dose of 1.15 and 2.30mg / kg respectively. Further these findings were supported and

confirmed by gross anatomical observation and histological examination. From the results of

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

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Volume 5, Issue 12, 905-916. Research Article ISSN 2278 – 4357

*Corresponding Author

Dr. N. Kabilan

Department of Siddha,

The Tamil Nadu

Dr.M.G.R. Medical

University, Chennai,

Tamil Nadu, India.

Article Received on

04 Oct 2016,

Revised on 24 Oct. 2016,

Accepted on 13 Nov. 2016

DOI: 10.20959/wjpps201612-8185


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the present investigation it was concluded that formulation like Poora parpam may serves as

a valuable lead for the clinical management of gastric ulcer.

KEYWORD: Anti-ulcer, Natural Poora parpam, Synthetic Poora parpam, Indomethacin,

Gastric ulcer, Ulcer index, Histopathology.

INTRODUCTION

The term peptic ulcer disease (PUD) generally refers to spectrum of disorders that includes

gastric ulcer (GU), pyloric channel ulcer, duodenal ulcer (DU) and postoperative ulcers at or

near the site of surgical anastomosis.[1]

By definition, they are defects in the mucosa that

penetrate at least into the sub mucosa, and often into the muscularis propria or deeper. Most

are round, sharply punched-out craters 2 to 4 cm in diameter; those in the duodenum tend to

be smaller, and occasional gastric lesions are significantly larger. At least 98% of peptic

ulcers are either in the first portion of the duodenum or in the stomach, in a ratio of about 4:1.

PUD are usually caused by H. pylori, reports from the US show that 30% of gastric ulcers

can be related to aspirin, indomethacin and other non-steroidal anti-inflammatory drugs

(NSAIDs). In developing countries, the ulcer groups are smaller and the gastric cancer group

may be larger. For example, in northern Brazil, gastric cancer is the most common

malignancy in men. Long-term use of NSAIDs is the second most common cause of ulcers,

and the rate of NSAID-caused ulcers is increasing. About 20 million people take prescription

NSAIDs regularly, and more than 25 billion tablets of over-the-counter brands are sold each

year in the U.S. alone. The most common NSAIDs are aspirin, ibuprofen and naproxen.

NSAIDs reduce prostaglandin biosynthesis by blocking COX. There are two forms of COX,

COX-1 and COX-2. Importantly, COX-1, but not COX-2, is expressed as the dominant,

constitutive isoform in gastric epithelial cells and is the major source of cytoprotective

prostaglandin formation. Inhibition of COX-1 at this site is thought to account largely for the

gastric ulcers.[2]

Gastric ulceration is a benign lesion on the mucosal epithelium upon exposure of the stomach

to excess acid and aggressive pepsin activity.[3]

It is the most prevalent gastrointestinal

disorder ever known, accounting for an estimated 15 mortality out of every 15,000

complications yearly in the world [4]

and [5]

.

Vatha gunmum (Peptic ulcer) is now believed to be due to an imbalance between the acids

and pepsin and defensive factors collectively called the mucosal barrier.[6]

Gastric ulcer is


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usually due to weakening of the gastric mucosa and duodenal ulcer due to the dominance of

acid and pepsin. Risk of ulcerogenesis is now greatly enhanced due to socio-economic

problems and exposure of man to many noxious agents and chemicals.[7]

Ulcer is the fourth

largest disease in Asia. Many drugs available on the market greatly reduce the morbidity and

mortality, but may also produce adverse reactions like gynaecomastia8 and also suffer from

high recurrence rates.

Numerous formulations indigenous to Indian system of medicine have been reported to be

helpful in successfully managing. Many Siddha formulations have been successful in

managing ulcer Poora parpam is one among them.

Pooram (Calomel) is one among the Panchasootham (five mercurial compounds) which is

widely used in Siddha preparation. The main component of the Pooram is Mercury. Mercury

is considered as Eesan in Siddha practice ie. Lord Siva performs all the three actions of

aakkal, kaaththal and azhiththal (creation, protection and destruction) through his different

incarnation. Mercury destroys almost all the diseases of mankind. The mercurial compound

has been in use in Siddha since many centuries. Pooram is identified and indicated for many

diseases like Iduppu Soolai (Lumbar spondilitis), Vatha gunmum (Peptic Ulcer), Suram

(Fever), Manjal Kamalai (Jaundice), Viranangal (Ulcers), Mega noigal (Venereal diseases),

Keel vatham (Osteo arthritis), Sirangu (Scabies) etc. (Thiyagarajan R (2004), “ Gunapadam

Part II (Thathu Jeeva vaguppu)”, Edition – IV, pp.283). The present study is an attempt to

compare the anti-ulcer potential of Poora parpam prepared from natural and synthetic

sources by using indomethacin induced ulcer model in rats.

MATERIALS AND METHODS

Experimental Animals

Healthy adult albino Wistar rats weighing between 150-175 g were used for the study. They

were purchased from Laboratory animal Medicine Unit, TANUVAS, Madhavaram Milk

Colony, Chennai - 600 051. The animals were housed in poly propylene cages and were kept

in well ventilated with 100% fresh air by air conditioning. A 12 hr light / dark cycle was

maintained .Room temperature was maintained between 20+2ºC and relative humidity 40–

65%. They were provided with food (Nutrilab Rodent feed, Provimi animal nutrition India

Pvt Ltd, Bangalore) and water ad libitum. All the animals were acclimatized to the laboratory

about 7 days prior to experimentation.


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The experimental protocol was approved by The Institutional Animal Ethics Committee of

National Institute of Siddha, Chennai, Tamil nadu, India.

Approval reference number- No.1248 / AC /09 / CPCSEA – 9 / DEC -2013 / 1 dated

05.12.2013

Animal Grouping

Animals were divided in seven groups of 6 animals each

GROUP I : 0.1% carboxy methyl cellulose (CMC) suspension (p.o)

GROUP II : 0.1% CMC suspension + Indomethacin 100 mg/kg b.w (p.o)

GROUP III : Ranitidine 100 mg/kg b.w + Indomethacin 100 mg/kg b.w (p.o)

GROUP IV :Natural Poora parpam 1.15 mg/kg b.w+ Indomethacin 100 mg/kg b.w (p.o)

GROUP V :Natural Poora parpam 2.30 mg/kg b.w+ Indomethacin 100 mg/kg b.w (p.o)

GROUP VI :Synthetic Poora parpam1.15 mg/kg b.w+ Indomethacin 100 mg/kg b.w (p.o)

GROUP VII :Synthetic Poora parpam2.30 mg/kg b.w+ Indomethacin 100 mg/kg b.w (p.o)

Experimental protocol for Indomethacin induced ulcer model in rats

After 12 h fasting, Group I were administered with 0.1% CMC suspension and Group II

(positive control): 0.1% CMC suspension + Indomethacin (100 mg/kg b.w).Group III with

Ranitidine (100 mg/kg), Group IV and V were administered with Natural Poora parpam at

(1.15 and 2.30 mg /kg b.w), respectively before 1 h of Indomethacin in 0.1% CMC

suspension (100 mg/kg, b.w), Group VI and VII were administered with Synthetic Poora

parpam at (1.15 and 2.30 mg /kg b.w ), respectively before 1 h of Indomethacin in 0.1%

CMC suspension (100 mg/kg,). All the test compounds were administered orally. 4 h after

Indomethacin administration, the animals were sacrificed by using excessive anesthesia. The

stomach was removed and opened along the greater curvature. The stomach was gently rinsed

with water to remove the gastric contents and blood clots. The inner surface of free stomach

was examined for gastric lesions. The number of ulcers was counted. Ulcer scoring was

carried out according to the method by as given below.[9]

The scores were:

0 = no ulcer

1 = superficial ulcer

2 = deep ulcer

3 = perforation


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Ulcer Index

Ulcer Index was measured by using the following formula.[10]

UI = UN + US + UP × 10−1

.

Where UI is the ulcer Index; UN is the average number of ulcers per animal; US is the average

number of severity score and UP is the percentage of animals with ulcers.

Percentage inhibition of ulceration [11]

Percentage inhibition of ulceration was calculated as follows:

× 100

There was a low percentage of ulcer in the study drug treated animals.

STATISTICAL ANALYSIS

The mean changes in Biochemical and Hematological parameters were statistically analyzed

by using Independent T Test and significant differences within groups were calculated using

the one way ANOVA test followed by Dunnett’s test to compare mean differences of control

and test drug treated groups. p ≤ 0.05 were considered statistically significant. The results

were expressed as the mean ± SD. Statistical analysis was performed using the SPSS version

18.

RESULT

Effect of Natural and Synthetic Poora parpam on Indomethacin Induced Ulcer in rats

Anti-ulcer activity of natural and synthetic Poora parpam was evaluated by using

Indomethacin induced ulcer in rats. In this study, it was observed that there was significant

increase in number of ulcer lesion and scoring of ulcer index in Indomethacin (100 mg/kg)

treated group. Treatment with natural and synthetic Poora parpam significantly reduced the

ulcer lesion index at dose level of dose of 1.15 and 2.30mg/kg. Similarly rats treated with

standard ranitidine (100 mg/kg) shown higher level of reduction in ulcer index when compare

to positive control group.

Standard drug ranitidine (100 mg/kg) treated group has shown highest percentage 69.9 %

inhibition of ulcer, whereas natural Poora parpam exhibit 38.43 and 57.99% inhibition at the

dose of 1.15 and 2.30mg/kg. Similarly rats treated with synthetic Poora parpam exhibit 29.12

and 48.21% inhibition at the dose of 1.15 and 2.30mg/kg. The results are tabulated in Table 1

and illustrated in Figure 1.


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Table 1: Effect of Natural and Synthetic Poora parpam on Indomethacin Induced Ulcer

Group Treatment and Dose

Indomethacin Induced Ulcer

Ulcer Index Percentage of

Ulcer Protection

I 0.1% carboxy methyl cellulose (CMC) suspension - 100

II Indomethacin 100 mg/kg b.w +0.1% CMC suspension 10.91±0.05* -

III Ranitidine 100 mg/kg b.w +Indomethacin 100 mg/kg b.w 3.27±0.04** 69.90

IV Natural Poora parpam 1.15mg/kg +Indomethacin 100 mg/kg b.w 6.71±0.03* 38.43

V Natural Poora parpam 2.30mg/kg +Indomethacin 100 mg/kg b.w 4.58±0.04* 57.99

VI Synthetic Poora parpam 1.15mg/kg+ Indomethacin 100 mg/kg b.w 7.73±0.02* 29.12

VII Synthetic Poora parpam 2.30 mg/kg +Indomethacin 100 mg/kg b.w 5.64±0.06* 48.21

Values of ulcer index are expressed as mean ± S.E.M. (N=6).Symbols represent statistical

significance: *p<0.05, **p<0.01, ***p<0.001. One way ANOVA followed by Dunnett’s test

Figure 1 :Effect of Natural and Synthetic Poora parpam on Indomethacin Induced Ulcer

Effect of Natural and Synthetic Poora parpam on Gross Anatomy of rat Stomach in

Indomethacin induced ulcer in rats.


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Gross anatomy of rat stomach treated with indomethacin 100 mg/kg b.w shown increases

number of ulcers with deeper projections and perforation when compare to control group

animals. Oral administration of natural and synthetic Poora parpam at the dose level of 1.15

and 2.30mg/kg, significantly reduces the severity of ulcer and restores the gastric mucosa

back to the normal level similar type of results were observed in animals treated with

ranitidine 100 mg/kg b.w which has significantly reduced the ulcer index and score. The

results are illustrated in Figure 2 and 3.

GROUP I GROUP II

GROUP III

Figure 2: Gross Anatomy of Rat Stomach belongs to Control, Indomethacin and

Ranitidine treated Group


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GROUP IV GROUP V

GROUP VI GROUP VII

Effect of Natural and Synthetic Poora parpam on Histopathological changes of rat

Stomach in Indomethacin induced ulcer in rats.

Figure 3: Gross Anatomy of Rat Stomach belongs to Natural and Synthetic Poora

parpam treated Group

Stomach histology of control group animals showed normal histology with intact epithelial

lining and gastric pits. Histopathological observation of stomach belongs to rats treated with

indomethacin 100 mg/kg b.w showed sever inflammatory changes in mucosa and sub mucosa

layers and also well differentiated ulceration of epithelia lining and mucosa layer in these

groups. Oral administration of natural and synthetic Poora parpam at the dose level of 1.15

and 2.30mg/kg, significantly reduced inflammatory changes and also restoration of

epithelium back to the normal. Animals treated with ranitidine 100 mg/kg b.w shows regular

arranged stomach layer and visible as mucosa layer, sub mucosa layer and muscularis propria

layer almost similar to that of the control group. The results are illustrated in Figure 4 and 5.


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GROUP I GROUP II

GROUP III

Figure 4: Histopathology of Rat Stomach belongs to Control, Indomethacin and

Ranitidine treated Group

GROUP IV GROUP V

GROUP VI GROUP VII

Figure 5: Histopathology of Rat Stomach belongs to Natural and Synthetic Poora

parpam treated Group


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DISCUSSION

Gastric ulcer is a common disorder of the digestive system. Current therapeutic regimens

largely rely on Western medicine. However, numerous studies have demonstrated that Siddha

system of traditional medicines can effectively treat gastric ulcer in humans and various

animal models via divergent mechanisms. The pathogenesis of NSAID-induced

gastrointestinal damage may also depend on prostaglandin-independent mechanisms, such as

uncoupling of oxidative phosphorylation, alterations of mucosal cell turnover as well as

neutrophil activation followed by enhanced endothelial adhesion 12

. These mechanisms, in

combination with those related to prostaglandin suppression, lead to micro vessel occlusion

and subsequent hyper production of reactive oxygen metabolites. Such substances are then

able to induce oxidative tissue injury which seems to play a prominent role in the

development of mucosal ulceration caused by NSAIDs.[13]

Treatment with natural and synthetic Poora parpam significantly reduced the ulcer lesion

index at dose level of dose of 1.15 and 2.30 mg/kg. Similarly rats treated with standard

ranitidine (100 mg/kg) shown higher level of reduction in ulcer index when compare to

positive control group. Standard drug ranitidine (100 mg/kg) treated group has shown highest

percentage 69.9 % inhibition of ulcer ,whereas natural Poora parpam exhibit 38.43 and

57.99% inhibition at the dose of 1.15 and 2.30 mg / kg . Similarly rats treated with synthetic

Poora parpam exhibit 29.12 and 48.21% inhibition at the dose of 1.15 and 2.30 mg / kg.

Gross anatomy of rat stomach treated with indomethacin 100 mg/kg b.w shown increases

number of ulcers with deeper projections and perforation when compare to control group

animals. Oral administration of natural and synthetic Poora parpam at the dose level of 1.15

and 2.30mg/kg, significantly reduces the severity of ulcer and restores the gastric mucosa

back to the normal level.

Histopathological observation of rat stomach shows that oral administration of natural and

synthetic Poora parpam at the dose level of 1.15 and 2.30mg/kg, significantly reduced

inflammatory changes and also restoration of epithelium back to the normal. Animals treated

with ranitidine 100 mg/kg b.w shows regular arranged stomach layer and visible as mucosa

layer, sub mucosa layer and muscularis propria layer almost similar to that of the control

group.


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CONCLUSION

Natural and synthetic Poora parpam at both dose levels shows significant decreases the

number of ulcers. It was observed that severity of ulcer lesion was reduced in the rat belongs

to treatment groups. From the study it was evident that formulation like Poora parpam may

serves as a valuable lead for the clinical management of gastric ulcer. But the underlying

mechanism of which the drug acts may be deeply investigated in near future.

ACKNOWLEDGMENT

The Director, National Institute of Siddha for permitted to carry out the entire study at

National Institute of Siddha, my dear PG students and The Noble research solutions, Chennai,

Tamil Nadu, India to assist me during the study.

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