Evaluation of Gastroprotective Effect of Aqueous Leaf Extract of Commicarpus chinensis in Pylorus Ligted Albino Rats.

Halkai M.A¹, Nitin Mahurkar², Akhil Ahmed M.D.¹*, Beknal A.K.¹, Sayeed Ul Hasan S.M.², Swathi M.¹

¹Department of Pharmacognosy, H.K.E.’s MTR IPS, Gulbarga – 585 105

²Department of Pharmacology, H.K.E.’s MTR IPS, Gulbarga – 585 105

ABSTRACT:

Objective

To evaluate the gastroprotective effect of aqueous extract of leaves of Commicarpus chinensis in pylorus ligated albino rats.

Materials and Methods

The plants were collected from the campus Gulbarga University, Gulbarga, Karnataka and authenticated by department of Botany, Gulbarga University, Gulbarga. The leaves were shade dried and subjected to extraction by percolation to obtain aqueous extract. The aqueous extract was evaluated for antiulcer activity by pylorus ligation model in albino rats. Albino rats of either sex weighing between 180-220 g were procured from the Central animal house, MR medical college, Gulbarga, Karnataka. The animals were divided into three groups of six animals in each. The animals were fasted for 48h and subjected to pylorus ligation technique for evaluating the antiulcer effect of C.chinensis. Prior approval of the IAEC was obtained for conduction of experiment.

Results

The aqueous leaf extract showed significant antiulcer activity on comparison with that of standard rabeprazole. The aqueous extract reduced the acidity of gastric juice and increased the pH of the gastric secretions, exhibiting statistically significant ulcer protection activity.

Conclusion

The antiulcer effect of C.chinensis may be due to antioxidant property contributed by the presence of tannins, flavonoids, saponins. Thus, C.chinesis possesses effective antiulcer property.

KEYWORDS: Commicarpus chinesis, Pylorus ligation, Rabeprazole, Antiulcer activity.

INTRODUCTION:

Medicinal plants are an important therapeutic aid for various ailments. In India, from ancient times, different parts of medicinal plants have been used to cure specific ailments. Today, there is widespread interest in drugs derived from plants. This interest primarily stems from the belief that green medicine is safe and dependable, compared with costly synthetic drugs that have adverse effects.^[1]

Commicarpus chinensis commonly known as “gum fruit” is a perennial herb belonging to the family Nyctaginaceae. The plant is widely distributed in South-eastern slopes of Western Ghats, Madurai district, Tamilnadu.^[2] Its Synonym is Boerhavia chinensis^[3] and its vernacular name is satodi. In Tamil, it is locally known as Piri kechoara.^[2]

The plant Commicarpus chinensis has been used in various ayurvedic and siddha preparations as hepatoprotective, gastroprotective, in down’s syndrome, as analgesic, antipyretic and anti-inflammatory agent.^[3]

Table.1: Gastroprotective effect of rabeprazole and C.chinensis on various GI biochemical parameters in pylorus ligated rats.

Groups	Volume of Juice (ml)	pH	Free acidity (mEq/l)	Total acidity (mEq/l)	Ulcer index	Percentage protection
Control	6.200±0.08	1.817±0.05	119.7±0.98	124.3±1.18	3.167±0.10	-
Rabprazole	1.450±0.05	5.267±0.10	64.90±0.79	84.00±1.23	0.500±0.12	84.2%
C.chinensis	2.150±0.05**	4.150±0.08**	75.33±1.40**	94.50±0.88**	0.916±0.08*	71.0%

Values are the mean±S.E.M, n= 6; *P<0.05 are significant on comparison of C.chinensis with rabeprazole

The whole plant of C.chinensis was evaluated for its antibacterial property, thus it is antibacterial in nature^[1]. In addition to the above mentioned properties the plant also possesses diuretic activity.^[4]The roots of the plant are used for its nutritional properties since it possess carbohydrates (starch),vitamins (niacin and ascorbic acid) and mineral.^[4]The plant also possesses antioxidant property^[5] and is used in treating skin diseases.^[2]

The present study mainly focuses upon evaluation of the gastroprotective property of C.chinensis in pylorus ligated albino rats using various biochemical parameters viz., volume of gastric juice (ml), pH, free acidity (mEq/L), total acidity (mEq/L), ulcer index and percentage protection.

MATERIALS AND METHODS:

The plant material was collected and authenticated by the department of Botany, Gulbarga University, Gulbarga, Karnataka. During the collection of plant material, care was taken so that the specimens to be studied were healthy, since microbial and other infections may change the metabolites produced by the specimens. After the collection, the plant was identified, authenticated and voucher specimen was deposited at departmental herbarium at Gulbarga University, Gulbarga, Karnataka for further reference.

Extraction Procedure

During percolation process, the plant material i.e. leaves were exhaustively extracted by fresh solvent (water). Only fresh water was used and the extraction consumes a large quantity of it. Re-percolation was performed, where the drug was first extracted with fresh solvent and then some of the percolate was used for exhaustive extraction by stage wise concentration in another percolator. Aqueous extract of the plant was obtained by percolation methods wherein care was taken to avoid over heating of material.

Percolation Procedure^[3]

Percolation is usually one of the most widespread-methods employed for plant extraction since it does not require much time. The equipment used was a conical glass container with a tap at the base of the apparatus used to set the rate of the solvent elution. Hot or cold solvent may be used. The sample was coarsely fragmented such that the particles were easily passed through a 3mm sieve. Particles of too large size may produce a high elution rate precluding the necessary equilibrium for the dissolution of the metabolites, and the menstrum (solvent) would percolate unsaturated. Since it is a continuous process in which the saturated solvent was constantly being displaced by fresh menstrum. Normally, percolation was not used as a continuous method because the sample was kept in solvent in the percolator for 24h (for three times), and then the extracted materials were collected and pooled. It has been observed that, after a triple-solvent extraction, the remaining marc does not contain valuable material.

Evaluation of Antiulcer activity^[6]

Wistar albino rats of either sex weighing between 180 – 220g were procured from the Central animal house, MR medical college, Gulbarga, Karnataka. The protocols were approved by IAEC.

The antiulcer effect of C.chinensis 500mg/kg b.w., p.o., was compared with reference drug rabeprazole 20mg/kg b.w., p.o. The ulcer index was calculated and other biochemical parameters like free acidity, total acidity, pH, volume of gastric juice, percentage protection were determined.

The animals were randomly selected and divided into 3 groups of 6 animals in each and were housed in separate perspex cages for a 12h light and dark cycle, allowing them to acclimatize with the laboratory conditions, during the whole period the animals were provided with sufficient food pellets and with water ad. libitum.

Following the 24h period of acclimatization the animals were fasted for a period of 48h. During this period the animals were placed in separate individual cages with raised mesh bottom in order to avoid coprophagy. After 48h of fasting the animals were administered with test and standard drugs, after one hour of drug administration the animals were anaesthetised under ether anaesthesia. The abdomen was opened with a mid line incision, the stomach was carefully lifted and the pylorus part was ligated following which the mid line incision was sutured and the animals were placed in separate cages for recovering from anaesthesia.

After 6h of pyloric ligation the animals were euthanesed under excess ether anaesthesia and the mid line incision was opened and stomach was dissected out. The gastric content of the stomach was collected in centrifugation tubes by making a small puncture in the stomach. The biochemical parameters like – free acidity, total acidity, ulcer index, pH of gastric juice, volume of gastric juice and percentage protection were evaluated.

The gastric contents were centrifuged at 1000 rpm for 10 min. The volume was noted. 1ml of supernatant liquid was pipette out and diluted to 10ml with distilled water (1:9). The pH of this solution was noted with the help of pH meter. The solution was titrated against 0.01N NaOH using Topfer’s reagent as indicator.

Titration was continued till the end point when the solution obtained an orange tinge. The volume of NaOH was noted which corresponds to free acidity. Titration was continued further following the addition of phenolphthalein indicator till the solution regains pink colour. This volume of NaOH corresponds to total acidity. Acidity was expressed as mEq/l/100gm. The acidity was determined using the following formula.

Acidity =

Vol. of NaOH x Normality x 100

0.1

The stomach was opened along the greater curvature and washed slowly under running tap water. The stomach was fixed on glass slide and was observed under 10X magnification for ulcers. The ulcers were scored as follows:

0.0 – Normal coloured stomach.

0.5 – red colouration.

1.0 – spot ulcers.

1.5 – Haemorrhagic streaks.

2.0 – ulcers ≥ 3 but ≤ 5

3.0 – ulcers > 5

Mean ulcer score for each animal was expressed as ulcer index.

RESULTS:

The aqueous leaf extract of C.chinensis significantly increased the pH (P<0.001) of the gastric content and reduced the volume (P<0.001), free acidity (P<0.001) and total acidity (P<0.001) significantly and provided significant gastro protection on comparison with the standard rabeprazole (table 1). The ulcer index (P<0.05) was found to be reduced significantly following C.chinensis treatment in comparison to rabeprazole. All the results were calculated using student t test (mann-whitney test) and were statistically significant. *P<0.05 was considered statistically significant.

DISCUSSION:

The peptic ulcer refers to painful sores or lesions present in the mucosal lining of the stomach (gastric ulcer) or duodenum (duodenal ulcer), which is the beginning of the small intestine. Peptic ulcer is thought to result from an imbalance between the aggressive (acid, pepsin, bile and H.pylori) and the defensive (gastric mucus and bicarbonate secretion, prostaglandins, nitric oxide, innate resistance of mucosal cells) factors.

Peptic ulcers are treated with drugs that kill the bacteria, reduce stomach acid, and protect the stomach lining. Antibiotics are used for eliminate H. pylori and two types of acid-suppressing drugs are used that include: H2 blockers and proton pump inhibitors.

The various causes of gastric ulceration lesions caused by stress, alcohol, H.pylori and due to use of NSAIDS have been shown to be mediated largely through generation of reactive oxygen species (ROS), especially the hydroxyl radical. A number of excellent drugs, developed over the decades have proven useful in controlling hyperacidity and ulceration but their long term use is reported to have serious side effects. Hence the investigations continue with an objective to find a herbal compound with more efficacy and lesser side effects.

In this study, an attempt was made to use an herbal drug i.e. C.chinensis which was evaluated for its gastroprotective effect, since, it possesses antioxidant^[3] property due to the presence of ascorbic acid, tannins, saponins and flavonoids in its leaves, hence the aqueous leaf extract was used for assessing the antiulcer property using pylorus ligation model. Being an antioxidant it can neutralize all the free radicals, nitrates, nitrites that causes cellular damage, at the same time it increased the gastric pH by reducing the acidity of gastric juice.

In order to determine the effectiveness of the extract, it was compared with a proton pump inhibitor i.e. rabeprazole, which exhibited maximum therapeutic effect by providing significant ulcer protection. Rabeprazole, being a potent proton inhibitor, decreases the excess acid secretion, by irreversibly blocking the H⁺,K⁺-ATPase of the parietal cells.

Thus both the leaf extract and the proton pump inhibitor were compared and the antiulcer activity was evaluated and it was found that C.chinensis possesses significant antiulcer effect that may be due to its antioxidant properties (Fig. 1).

CONCLUSION:

C.chinensis exhibited significant gastro protection by increasing the pH, reducing the acidity and decreasing the ulcer index thereby, exhibiting significant gastric protection. Thus, it is an effective gastroprotective agent and can be used as antiulcer drug.

ACKNOWLEDGEMENT:

The authors are thankful to authorities of HKE society and HKES’s Matoshree taradevi rampure institute of pharmaceutical sciences, Gulbarga, Karnataka for providing the necessary facilities.

REFERENCES:

1) Nair R, Chanda S. Activity of some medicinal plants against certain Pathogenic bacterial strains. Indian J Pharmacol. 2006; 38(2): 142-4.

2) Vanila D, Ghanthikumar S, Manickam VS. Ethnomedicinal uses of plant in the plains area of tirunelveli - district, tamilnadu, india. Ethnobotanical leaflets 12. 2008; 1198-1205.

3) Boxi M, Rajesh Y, Kumar RV, Praveen B, Mangamma K. Extraction, phytochemical screening and in vitro evaluation of antioxidant properties of Commicarpus chinensis (aqueous leaf extract). Int J Pharma Bio Sci 2010; 1(4): 537- 47.

4) Harvey S.K. A preliminary experimental study of the diuretic activity of some indigenous drugs. Indian J Med Res. 1966; 54(8): 774-78.

5) Mohan VR, Chinnamadasam. Nutritional and antinutritional evaluation of some unconventional wild edible plants. Tropical and Subtropical agroecosystems 2010; 12: 495-506.

6) Kulkarni SK. Handbook of Experimental Pharmacology 3^rdEd. Vallabh Prakashan, New Delhi. 2005: 148-50.

Received on 02.07.2012

Modified on 27.07.2012

Accepted on 06.08.2012

Research J. Pharmacology and Pharmacodynamics. 4(5): September –October, 2012, 307-310