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Research Article ISSN: 0974-6943

A.K. Manna et al. / Journal of Pharmacy Research 2009, 2(5),785-788

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The antiulcer activity of Pterospermum acerifolium bark extract in experimental animal


A.K. Manna**, AK Behera, J Jena, S Manna, S Karmakar*, Dr.S Kar**, B.R.Panda**, S. Maity** *Dept. of pharm Tech. Jadavpur University, kolkata -700032. **Seemanta Institute of pharmaceutical sciences, Jharpokharia, Orissa. 757086. Received on: 27-09-2008; Accepted on: 28-03-2009 ABSTRACT The role of ethanolic fraction of Pterospermum acerifolium bark extract on different experimental ulcer models in rats was investigated.The extract demonstrated significant antiulcer activity against aspirin, indomethacin & ethanol induced ulcerations, significant inhibition of gastric secretary volume, and total acidity in pylorus ligated rats were observed to occur with the extract. Keywords: Pterospermum acerifolium, ulcer , Leukotrienes INTRODUCTION Pterospermum acerifolium wild (Sterculiaceae) commonly known as Kanak champa is a shurbs distributed in tropical Asia. It has been traditionally used for blood troubles, inflammation, ulcer, tumors, leprosy and for small pox eruptions (wealth of India).In an earlier study in our laboratory, the ethanolic extract of P. acerifolium was found to possess anti-inflammatory activity.The work was aimed at the scientific validation of the ethnopharmacological calim about anti ulcer properties of the bark extract.Earlier studies in our laboratory have revealed that the ethanolic fraction of Pterospermum acerifolium bark extract possesses a significant anti-inflammatory and analgesic property.6 However, our studies revealed that Pterospermum acerifolium possess significant antiulcer activity against various experimental models of gastro intestinal ulceration in rat. The present study was undertaken to evaluate the effect of Pterospermum acerifolium on different experimental models of gastro intestinal ulceration. MATERIALS & METHODS: Plants materials P. acerifolium bark were collected from East Midnapur (West Bengal, India) in July 2007 and authenticated by comparison with a voucher. Specimen in Botanical Survey of India, Kolkata.One kg. of the air dried barks were blended to a fine powder and extracted with Pet. Ether, chloroform and ethanol for 6 days (144hours). The extract was concentrated using a rotavapor. The extract was dissolved in normal saline before orally administrating 150 and 300 mg /kg of the extract to the rats. The preliminary investigation for the antiulcer activity

According to Shay and Sun,21 the genesis of peptic ulcer mechanism is related to the balance between the defensive and aggressive factors. Gastric mucosa is an active area of arachidonic acid metabolism (where both cyclo oxygenase and lipoxygenase enzyme are involved) the integrity of gastric mucosal barrier is influenced by mucus secretion, acid secretion, and gastric blood flow. 18 The gastric Phytochemical screening: irritance of non-steroidal anti-inflammatory drugs (NSAIDs) on the gastric mucosa is one of the major disadvantage of their using inflamThe extract and its fraction were tested by the librman Burchard, matory. It is well established that NSAIDs causes inhibition of the Ferric chlorides, Magnesium tracings and Vanillin sulphuric acid tested synthesis of cytoprotective prostaglandin. to determine the presence of sterols phenolic compound, flavonoids and saponins respectively. However, according to Cohn4 NSAIDs increase the susceptibility of gastric mucosa to injury by gastric acid bile salt and ethanol. Chemicals: As it is essential to evaluate the ulcerogenic potential of any antiinflammatory agent, the same was evaluated with ethanolic fraction Indomethacin (sigma), aspirine (sigma) and all the chemical collected of Pterospermum acerifolium bark extract and it is found that from Sisco laboratory Mumbai. Pterospermum acerifolium did not possesses ulcerogenic activity. Plant material: *Corresponding author. E-mail: amanna73@yahoomail.com Authenticated bark of Pterospermum acerifolium were air dried powdered and subsequently extracted with petroleum ether, chloroform and ethyl alcohol in a soxhlet extractor. The ethanol extract was dried in vacuum and yellowish brown residue was obtained. Just prior Journal of Pharmacy Research Vol.2.Issue 5.May 2009
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A.K. Manna et al. / Journal of Pharmacy Research 2009, 2(5),785-788 to testing, the extract was dissolved in distil water which serve as the et al.5 Gastric juice was diluted (1:250) with 0.01N HCL and 1 ml of vehicle in our experiments. diluted gastric juice was taken for further estimation, which was added to 2.5ml of 2% haemoglobin solution in 0.06M HCL. The minture was Animals used: incubated at 370C for 20min. and thereafter 0.6m trichloro acitic acid added. The tubes were kept in an ice bath for 15min. and then centriAdult Charles Foster rats (of male 140-160g) house and norTable-1. Effect of Pterospermum acerifolium extract on Aspirin induced mal laboratory condition (24 + 20C) were used. They were supplied gastric ulceration (values are expressed as mean + S.E.; n = 6) (Dose of with standard food pellets (water ad libitum). The animal were fasted Pterospermum acerifolium extract (T1 150 mg/kg and T 2 = 300 mg/kg) = for 24 hours prior to experimentation, however they were supplied standard drug omeprazole = S = 10 mg/kg) with water adlibitum till 1 hour before commencement of the experiAnimal No. Ulcer Index mentation, how ever they were supplied with water ad libitum till one Control T1 T2 S hour before commencement of experimentation. Aspirin induced gastric ulcer: Animals were divided into three groups (n = 6) Pterospermum acerifolium (150, 300mg/kg) inter peritoneal and control vehicle were administered 30 min. before the administration of aspirin (400mg/kg) per oral [PO]. The animals were scarified, after 6 hours following the administration of aspirin, stomachs were removed and 2% formalin was injected into the stomach. The stomach was open along with greater curvature and immersed in 2% formalin solution. The length of each lesion was measured under the dissecting microscope (X 10). The sum of the length (mm) of all lesions for each rat were used in lesion index. 1, 12 Indomethacin induced ulceration:
1 2 3 4 5 6 Mean + S.E. % Inhibition 3.5 3.2 3.1 4.2 4.5 3.8 3.71 0.095 1.9 1.7 1.2 1.5 1.3 1.4 1.5 0.047* 59.66 0.4 0.2 0.8 0.4 0.5 0.3 0.43 0.008* 88.40 0.1 0.2 0.3 0.2 0.2 0.3 0.21 0.007* 94.33

P values vs. control (by studentt test) * p <0.05

Ulcer index

4 2 * 0
con .. T2 T1 S
S 1.2

Male rats tested for 24 hours administered with Pterospermum acerifolium (150mg/kg) [PO] or the control vehicle and 30min. later, indomethacin (20mg/kg) [PO] was administered. The animals were sacrificed 18 hours later of the ulcer index was determined P values vs. control (by studentt test) * p <0.05 microscopically.17,10 Fig.1.Effect of Pterospermum acerifolium extract on Aspirin induced gastric ulceration (values a* re expressed as mean + S.E.; n = 6) Ethanol induced ulceration: (Dose of Pterospermum acerifolium extract (T1 150 mg/kg and T2 = 300 mg/kg) = standard drug omeprazole = S = 10 mg/kg Male Albino rats were divided into three groups (n = 6) Pterospermum acerifolium (150, 300mg/kg) and control vehicle was Table-2. Effect of Pterospermum acerifolium extract on Alcohol induced administered [PO] after 30 min., ethanol (50%, 5ml/kg) was adminis- gastric ulceration (values are expressed as mean + S.E.; n = 6) (Dose of tered orally. The animals was sacrificed after one hour and the lesion Pterospermum acerifolium extract (T1 150 mg/kg and T 2 = 300 mg/kg) = standard drug omeprazole = S = 10 mg/kg index was determined microscopically.18
Animal No.

Gastric secretory study in Pylorus ligated rats:


1

Ulcer Index Control T1 8.1 5.1

T2 2.2

The effect of Pterospermum acerifolium (150, 300mg/kg) on 2 8.3 6.8 2.8 1.8 8.2 6.1 3.6 1.4 gastric secretion (volume, pH, total acidity and peptic activity) was 3 7.6 5.6 3.1 1.3 studied in Pylorus ligated rats, Pterospermum acerifolium was ad- 4 5 7.2 5.4 3.8 1.7 ministered orally 30 min. pyloric ligation. The animals were sacrificed 6 8.4 6.2 2.7 1.1 7.9 5.86 3.03 1.41 after 4 hours following pyloric ligation,1 gastric content were con- Mean 0.481 0.066** 0.099* 0.038* trolled and analyzed for volume, acidity ,pH and peptic activity. For S.E. % Inhibition 25.82 61.65 82.15 determination of acidity, 0.2ml of gastric juice was taken and diluted to 2ml with distil water in small conical flask; 1 drop of 1% phenopthalin P values vs. control (by studentt test)* p <0.01),** p <0.05 was then added to it. The total acidity was determined by tritrating with 0.01N NaOH and was expressed in meq/L/hr.Peptic activity was fuged. 1ml of supernatant was used to determine the concentration of 9 determined by the modified Anson method2 as described by Debnath liberated amino acids by the lowry et al . Journal of Pharmacy Research Vol.2.Issue 5.May 2009
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A.K. Manna et al. / Journal of Pharmacy Research 2009, 2(5),785-788


Ulcer index 10 8 6 4 2 0
co ntr ol

acerifolium was very little protection of gastric mucosa (Table-3).


** *

Ethanol induced gastric ulceration:


1 2 St d.

Pterospermum acerifolium (150, 300 & 400mg/kg) showed significant inhibitory activity against ethanol induced gastric ulcer* ation. Pterospermum acerifolium at a dose of 150mg/kg showed very P values vs. control (by student t test)* p <0.01),** p<0.05 little protective activity and there was considerable damage of the Fig.2. Effect of Pterospermum acerifolium extract on Alcohol induced mucosal layer accompanied with diffused hyperaemia and glandular gastric ulceration (values are expressed as mean + S.E.; n = 6) (Dose of disruption (Table-2).
T

Table-3. Effect of Pterospermum acerifolium extract on Indomethacin (20mg/kg) induced gastric ulceration (values are expressed as mean + S.E.; n = 6) (Dose of Pterospermum acerifolium extract (T1 150 mg/kg and T 2 = 300 mg/kg) = standard drug omeprazole = S = 10 mg/kg)
Animal No. 1 2 3 4 5 6 Mean S.E. % Inhibition Ulcer Index Control T1 2.6 2.8 2.1 2.2 2.3 2.7 2.4 0.118 28.97 2.1 2.2 1.4 1.3 1.9 1.7 1.7 0.15** 64.08 T2 0.8 0.9 0.5 1.1 0.8 1.2 0.88 0.101* 81.63 S 0.4 0.2 0.2 0.9 0.6 0.4 0.45 0.074*

Ulcer index(in mm)

Pterospermum acerifolium extract (T1 150 mg/kg and T 2 = 300 mg/kg) = standard drug omeprazole = S = 10 mg/kg)

3 2 1 0

**

P values vs. control (by student t test)* p <0.01),** p<0 Fig.3.Effect of Pterospermum acerifolium extract on Indomethacin (20mg/kg) induced gastric ulceration (values are expressed as mean + S.E.; n = 6) (Dose of Pterospermum acerifolium extract (T1 150 mg/kg and T2 = 300 mg/kg) = standard drug omeprazole = S = 10 mg/ kg Table 4. Effect of Pterospermum acerifolium extract on Gastric secretion following Administra-tion of 50% V/v alcohol (5 ml/kg; P.O) pylorus ligated rats [Dose of Pterospermum acerifolium extract T1 150 mg/kg; T2 300mg/kg; S-10mg/kg P.o]
Animal no Volume (ml/100gm) C T1 T2 S PH C T1 T2 S Total acidity (Meq/lit/hr) C T1 T2 Peptic activity mg of tyrosine/ml C T1 T2

1. 1.8 1.7 1.4 1.3 4 5 5 5 42 37 31 28 16.42 11.13 8.71 2. 1.7 1.6 1.4 1.2 4 5 6 6 42 30 28 27 16.67 14.23 6.12 3. 1.6 1.5 1.3 1.3 4 4 6 5 39 31 30 26 17.64 14.18 9.52 4. 1.9 1.7 1.3 1.2 5 5 5 5 38 33 29 29 15.12 15.54 10.6 5. 2.1 1.8 1.4 1.4 6 5 5 5 39 30 28 28 16.47 14.16 8.72 6. 2.1 1.6 1.4 1.3 4 5 6 5 36 33 29 27 17.12 13.61 9.22 Mean 1.87 1.65 1.37 1.28 4.5 4.83 5.5 5.1 39.7 32.7 29.1 27.6 16.57 13.64 8.8 S.E.M .08 .04 .02 .03 .34 .17 .23 .17 .93 .59 .30 .34 .35 .51 .61 %inhibition 13.15 36.58 45.52 6.89 18.2 12.9 21.3 36.4 43.5 21.5 88.1 Remarks: *Significant decrease of volume, total acidity, and peptic activity. The PH of gastric content remained unaffected in pylorus ligated rats.

l ntro Co

T1
S

T2

. Std

RESULTS: Gastric secretory study following pylorus ligation in rats: Aspirin induced gastric ulcer: Pterospermum acerifolium (150, 300mg/kg) significantly inhibited gastric ulceration in rats (table 1).There was significant inhibition of ulcer formation with bark extract gastric mucosa retained almost normal appearance with the higher dose of Pterospermum acerifolium (300 & 400mg/kg). Indomethacin induced gastric ulceration: The risk of gastric ulceration is a predominant factor which reSignificant inhibition of ulceration was observed with bark stricts the use of NSAIDs in a variety of inflammatory condition. Studies extract (150, 300 & 400mg/kg). However at lower dose of Pterospermum with several neutral products in our laboratory reveals that plants Journal of Pharmacy Research Vol.2.Issue 5.May 2009
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The bark extract at the dose of(300mg /kg) significantly decreased gastric volume and total acid contain pH remain unchanged (Table-4).However, no significant change could be observed on the peptic activity in the groups pretreated with Pterospermum acerifolium (300mg/kg). DISCUSSION:

A.K. Manna et al. / Journal of Pharmacy Research 2009, 2(5),785-788 such as Brophyllumpinatum, 14 P. indica19,20 etc. possess significant din. In: Prostaglandin and Leukotrienes in Gastrointestinal disanti inflammatory and anti- ulcer activity. ease. Berlin: Springer Verlag : 1988. p. 148-51.
5.

Studies with ethanolic fraction of Pterospermum acerifolium root extract initially reveals that anti-inflammatory activity may be due to inhibition of arachidonic acid metabolism. Aspirin and indomethacin causes gastro intestinal ulceration by inhibition of gastric mucosal biosynthesis of cytoprotective prostaglandin PGE2 and PGI2.16 It may also be mentioned that the inhibition of prostaglandin biosynthesis may lead to gastric hyper secretion, the damage of gastric mucosa may occur due to the direct action of these drug and thereby leading to mast cell degranulation and release of histamine NSAIDs cause shifting of arachidonic acid metabolism to the 5-LO pathway thereby leading to over production of leucotrienes.18 Our studies with Pterospermum acerifolium showed that it was capable of inhibiting aspirin and indomethacin induced gastric ulceration in rats. Ethanol causes extensive ulceration in fasted animals. Such ulceration may be attributed to stasis of gastric blood flow, thereby leading to haemorrhage and necrosis. Early studies reveals that 5-lo product gastric mucosa from ethanol induced ulceration.10 Thus studies have also shown that intragastric administration of ethanol causes increased generation of LTC4 resulting in mucosal and sub-mucosal vasoconstriction of sub-mucosal micro-vessels and tissue necrosis.15 LTD4 has been found to decrese trans gastric potential.16 All these factors effectively contribute to gastric ulceration and damage. Further investigation on gastric secretion in pylorus ligated rat, p acerifolium extract produce significant decrease in gastric volume, total acidity and peptic activity of gastric juice in animal treated with alcohol after pylorus ligation. And pH of gastric juice was found to be unaffected in this contest it may mentioned that ethanol administration probably induces LTD4 production their by leading to increases gastric protein contained and decreases transgastric potential (Sen et al 2002). Thus the possible reasons of the antiulcer effect of P.acerifolium may be due to (i) the inihibition of 5-LO enzyme (ii) blockade of LTC4, LTD4 synthesis (iii) generation of free radicals; and/or (iv) inhibition of histamine release following mast cell degranulation. One or more of the mentioned reasons may be responsible for the antiulcer activity P.acerifolium. Further work in this regard may be done.
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Source of support: Nil, Conflict of interest: None Declared

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