Anti-ulcer activity of Cleome viscose linn. against gastric ulcer in rats

Poonia Lalita*, Singh Gajendra Kamal, Nagori Badri Prakash

Lachoo Memorial College of Science and Technology (Autonomous) Pharmacy Wing, Jodhpur, Rajasthan India

ABSTRACT:

Cleome viscosa linn. (Capparidaceae) is also known as Tickweed, or Spider plant. In Asia and Africa the leaves and seeds used as a rubefacient and vesicant and to treat infections, fever, rheumatism and headache. A decoction is used as an expectorant and digestive stimulant and the vapour from a steaming decoction of the whole plant is inhaled to treat headache. The seeds and its oil have antihelminthic properties but they are ineffective in treating roundworm infections. The roots are a remedy for scurvy and rheumatism. An aqueous seed extract displayed significant analgesic activity in mice and local anaesthetic activity in guinea pigs. In tests with rats the anti-diarrhoeal and antipyretic. The whole herb is used in treatment of inflammation of the middle ear and applied on wounds and ulcers, so the research was to prove experimentally anti ulcer activity in whole plant of Cleome viscosa linn. The effect of alcoholic extract of Cleome viscosa linn. was investigated in rats to evaluate the anti-ulcer activity by using pyloric ligation model experimentally induced gastric ulcer. The parameters taken to assess anti-ulcer activity were volume of gastric secretion, pH, free acidity, total acidity and ulcer index. The results indicate that the alcoholic extract significantly (P < 0.05) increases pH and decreases the volume of gastric acid secretion, free acidity, total acidity and ulcer index with respect to control.

KEYWORDS: Cleome viscosa linn., anti-ulcer, free acidity, pyloric ligation, ulcer index

INTRODUCTION:

Peptic ulcer is the most common gastrointestinal disorder in clinical practice. Considering the several side effects (arrhythmia’s, impotence, and haematopoeitic changes) of modern medicine¹, herbal plant drugs possessing fewer side effects should be looked for as a better alternative for the treatment of peptic ulcer. There is evidence concerning the participation of reactive oxygen species in the etiology and pathophysiology of human diseases, such as neurodegenerative disorders, inflammation, viral infections, autoimmune pathologies and digestive system disorders such as gastrointestinal inflammation and gastric ulcer². The study assumes significance in the context that prolonged use of synthetic anti-ulcer drugs leads to adverse drug reactions and a search for new anti-ulcer agents that retain therapeutic efficacy and are devoid of adverse drug reaction is warranted. A study of the efficacy of an alcoholic extract of Cleome viscose linn. in gastric ulcer with pylorus ligation model was undertaken in a rat model.

Cleome viscosa Linn. (Capparidaceae) is also known as Tickweed, or Spider plant. It occurs in woodland and grassland, and is a weed of fallow land, fields, roadsides and wasteland, often occurring on sandy soils, but sometimes on calcareous and rocky soils.

In Asia and Africa the leaves and seeds used as a rubefacient and vesicant and to treat infections, fever, rheumatism and headache. The whole herb is used in treatment of inflammation of the middle ear and applied on wounds and ulcers. A decoction is used as an expectorant and digestive stimulant and the vapour from a steaming decoction of the whole plant is inhaled to treat headache³. The seeds and its oil have antihelminthic properties but they are ineffective in treating roundworm infections⁴. The roots are a remedy for scurvy and rheumatism⁵. An aqueous seed extract displayed significant analgesic activity in mice and local anaesthetic activity in guinea pigs⁶,⁷. In tests with rats the anti-diarrhoeal⁸ and antipyretic⁹ activities of the extracts have been confirmed. Hence in the present study, the Cleome viscose linn. plant has been selected to investigate the anti-ulcer study.

MATERIALS AND METHODS:

Plant material:

Collection of whole plant of the Cleome viscose linn. was done personally from the nearby area of Umed palace, Jodhpur (Raj.) in the month of July-september 2010. Taxonomical identification and authentication of the plant was done by Dr. P. J. Parmar, Joint Director, Botanical Survey of India, Arid Zone Regional Center, Jodhpur (Raj.)

Whole plant material was air dried at room temperature under shade and the dried parts were then ground to coarse powder with the help of suitable grinder. The powdered plant material was then kept in airtight polythene bags and stored in cool and dark place to avoid deterioration by elevated temperature, light and moisture.

Extract Preparation:

The whole plant was shade-dried and made into a coarse powder which was passed through a 40-mesh sieve to get a uniform particle size and then used for extraction. A weighed quantity (200 g) of the powder was then subjected to continuous hot extraction in Soxhlet apparatus with 450 ml ethanol after defattation with pet ether and the residual marc was collected. The extract was filtered through a cotton plug, followed by whatman filter paper (no.1). The extract was evaporated under reduced pressure using a rota evaporator at a low temperature (40-60^oC) until all the solvent had been removed to give an extract sample with a yield of 3.2% w/w in relation to the dried starting material. Preliminary phytochemical analysis was carried out to identify presence of phytoconstituents in the crude alcoholic extract. ¹⁰

Preliminary phytochemical analysis:

The alcoholic extract of Cleome viscose linn. was then subjected to preliminary phytochemical ¹¹ analysis to assess the presence of various phytoconstituents, it revealed that the presence of alkaloids, steroids, flavonoids, glycosides and tannins. Preliminary Thin layer chromatography studies also confirmed these constituents ¹².

Animals:

Wistar albino rats weighing 150-200g of either sex maintained under standard husbandary conditions (temp 23±2oC, relative humidity 55±10% and 12 hours light dark cycle) were used for the screening. Animals were fed with standard laboratory food and ad libitum during the study period. The experimental protocol has been approved by institutional animal ethics committee.

Pylorus- ligation induced gastric ulcer:

Either sex of albino rats weight 150-200g were selected for modified pyloric ligation ulcer model according to Goel et al. (1985) ¹³. Rats were divided into five groups, each group consisting of six animals. Animals were fasted for 24 h before experiment. One group received normal saline 2 ml/kg (negative control), the second group received Ranitidine 100 mg/kg by oral route (positive control) and the third, fourth and fifth groups received alcoholic extract Cleome viscose linn. (200, 400.0, 600.0 mg/kg respectively) by oral route, 3 hour prior to aspirin administration. Treatment was continued for 3 days and pylorus was ligated on fourth day under ether anaesthesia. The abdomen was closed and the animals were left to recover. Animals were sacrificed 4 h later with over dose of ether and the stomach was opened to collect the gastric contents. The total volume of gastric content was measured. The gastric contents were centrifuged at 5000 rpm for 5 min. One ml of the supernatent liquid was pipetted out and diluted to 10 ml with distilled water. The solution was titrated against 0.01N NaOH using Topfer’s reagent as indicator, to the endpoint when the solution turned to orange colour. The volume of NaOH needed was taken as corresponding to the free acidity. Titration was further continued till the solution regained pink colour¹³. The volume of NaOH required was noted and was taken as corresponding to the total acidity.

Acidity was expressed as:

Acidity = (Volume of NaOH x Normality of NaOH x 100 mEq/l) /0.01

Ulcer index (UI):

The mucosa was flushed with saline and stomach was pinned on frog board. The lesion in glandular portion was examined under divider a 10 X magnifying glass and length was measured using a divider and scale and gastric ulcer was scored. Ulcer index of each animal was calculated by adding the values and their mean values were determined.¹⁴

0 - normal coloured stomach.

0.5 - Red colouration.

1 – Spot colouration.

1.5 – hemorrhagic streak.

2 – Ulcers.

3 – Perforations.

Statistical analysis:

The values Mean±SEM were calculated for each parameter. For determining the significant inter group difference each parameter was analysed separately and one-way analysis of variance¹⁵ (Gennaro, 1995) was carried out and the individual comparisons of the group mean values were done using Dunnet’s test ¹⁶.

RESULT & DISCUSSION:

Preliminary phytochemical screening revealed the presence of alkaloids, steroids, flavonoids, glycosides and tannins. In Pylorus ligation induced ulcer model alcoholic extract of the Cleome viscose linn. in the doses of 400 and 600 mg/kg produced a reduction in the ulcer index, gastric volume, free acidity, total acidity and raised gastric pH significantly in comparison with control group. Ranitidine reference drug produced significant reduction gastric ulcer and total acid output as compared to control group (Table 1). The control animals had ulcers and haemorrhagic streaks, whereas in animals administered with the extracts of Cleome viscose linn.showed significant reduction in ulcer index (P < 0.05) (Figure 1).

Figure 1

(a) Stomach of control rat;

(b) Standard drug treated

(c) Effect of alcoholic extract(dose-200 mg/kg) on aspirin induced gastric ulcer in rat;

(d) Effect of alcoholic extract(dose-400 mg/kg) on aspirin induced gastric ulcer in rat;

(e)Effect of alcoholic extract (dose-600 mg/kg) on aspirin induced gastric ulcer in rat;

Table 1:

Treatment	Dose(mg/kg)	pH	Vol. of gastric juice(ml)	Free acidity (mEq/L)	Total acidity(mEq/L)	Ulcer index
Control (Normal saline)	2 ml/kg	3.9 ±0.341	2.26 ±0.298	27.03 ±0.882	70.06 ±2.481	6.16 ±2.460
Standard (Ranitidine)	100 mg/kg	5.51 ±0.367	2.03 ±0.188	10.33 ±0.442	22.26 ±1.241	2.583 ±0.533
Alcoholic extract	200 mg/kg	3.16±0.849	2.08±0.279	25.83±1.502	37.46±1.308	1.91±0.786
	400 mg/kg	5.16±0.811	1.90±0.191	20.0±2581	32.23±1.202	0.75±0.629
	600 mg/kg	5.33±0.657	1.41±1.106	24.5±1.644	34.23±1.22	2.083±0.975

Results are mean ± S.E.M. (n = 6). Statistical comparison was performed by using ANOVA coupled with student‘t’ test. P<0.05 were consider statistically significant when compared to control group

The anti-ulcer activity of the plant of Cleome viscose linn. was evaluated by employing pylorus ligation ulcer models. This model represent some of the most common causes of gastric ulcer in humans. Many factors and mechanisms are implicated in the ulcerogenesis and gastric mucosal damage induced by pylorus ligation model employed in the present study involving, depletion of gastric wall, mucin mucosal damage induced by non-steroidal anti-inflammatory drugs and free radical production ¹⁷. The extract of Cleome viscose linn. has significantly protected the gastric mucosa against pylorus ligated model challenge as shown by reduced values of lesion index as compared to control group suggesting its potent cytoprotective effect. It has been proposed that in pyloric ligation, the digestive effect of accumulated gastric juice and interference of gastric blood circulation are responsible for induction of ulceration ¹⁸. The antiulcer activity of Cleome viscose linn. extract in pylorus ligation model is evident from its significant reduction in gastric volume, total acidity, free acidity, ulcer index and increase in pH of gastric juice. Because of animals treated with Cleome viscose linn. extract significantly inhibited the formation of pylorus ulcer in the stomach and also decreased both acid concentration, gastric volume and increased the pH values. It is suggested that Cleome viscose linn. extract can suppress gastric damage induced by aggressive factors. It is generally accepted that gastric ulcers result from an imbalance between aggressive factors and the maintenance of the mucosal integrity through endogenous defence mechanisms. The preliminary phytochemical studies revealed the presence of flavonoids in alcoholic extract of Cleome viscose linn.; various flavonoids have been reported for its anti-ulcerogenic activity with good level of gastric protection ^{19, 20}. So the possible mechanism of antiulcer action against gastric ulcer of Cleome viscose linn. may be due to its flavonoid content.

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Received on 25.01.2013

Modified on 05.02.2013

Accepted on 09.02.2013

Research Journal of Pharmacognosy and Phytochemistry. 5(3): May-June 2013, 115-118