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 Table of Contents  
ORIGINAL ARTICLE
Year : 2015  |  Volume : 3  |  Issue : 1  |  Page : 12-17

Evaluation of analgesic, anti-inflammatory, antipyretic and antiulcer effect of aqueous and methanol extracts of leaves of Polygonum minus Huds. (Polygonaceae) in rodents


1 Unit of Pharmacology, Faculty of Pharmacy, AIMST University, Bedong; School of Pharmaceutical Sciences, University Sains Malaysia, Penang, Malaysia
2 Unit of Pharmacology, Faculty of Pharmacy, AIMST University, Bedong, Malaysia
3 School of Pharmaceutical Sciences, University Sains Malaysia, Penang, Malaysia

Date of Web Publication13-Apr-2015

Correspondence Address:
Parayil Varghese Christapher
Unit of Pharmacology, Faculty of Pharmacy, AIMST University, Bedong - 08100
Malaysia
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2321-4848.154919

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  Abstract 

Background: Polygonum minus (Kesum) is an annual plant that grows throughout South East Asian countries. The Leaf of P. minus is commonly used as diet ingredient in Malaysia. Traditionally the decoction of leaves of this plant is used to treat stomach ache and digestive problems. The plant has known antioxidant activity, and its pharmacological properties are remaining unclear. Hence the study is planned to evaluate the analgesic, anti-inflammatory, antiulcer and antipyretic activity of kesum. Materials and methods: P. minus leaves was extracted with methanol and distilled water by simple maceration. The dried extract was used for further phytochemical and pharmacological analysis. The analgesic effect of methanol and aqueous extract of P. minus was studied using acetic acid, tail immersion and formalin induced pain in rats. The anti-inflammatory effect of both extracts was studied using carrageenan induced paw edema in rats. The pyloric ligation model was used to study the antiulcer effect. The antipyretic effect was studied using Brewer's yeast induced pyrexia. Results: The percentage yield of aqueous and methanol extract of P. minus leaves were 1.15 and 2.57% w/w respectively. Both the extract showed significant analgesic effect against acetic acid writing, tail immersion and formalin induced pain methods, but the effect was not equivalent to that of standard. Aqueous extract showed significant anti-inflammatory action and methanol extract showed significant anti-ulcer effect. Conclusion: The aqueous extract of the P. minus has significant analgesic and anti-inflammatory action, whereas methanolic extract showed presence of analgesic and anti-ulcer activity. Both aqueous and methanolic extract did not show any significant antipyretic activity.

Keywords: Analgesic, anti-inflammatory, anti-ulcer, Polygonum minus


How to cite this article:
Christapher PV, Xin TY, Kiun CF, Leng LC, Fu NG, Yuan GL, Parasuraman S, Vikneswaran M. Evaluation of analgesic, anti-inflammatory, antipyretic and antiulcer effect of aqueous and methanol extracts of leaves of Polygonum minus Huds. (Polygonaceae) in rodents. Arch Med Health Sci 2015;3:12-7

How to cite this URL:
Christapher PV, Xin TY, Kiun CF, Leng LC, Fu NG, Yuan GL, Parasuraman S, Vikneswaran M. Evaluation of analgesic, anti-inflammatory, antipyretic and antiulcer effect of aqueous and methanol extracts of leaves of Polygonum minus Huds. (Polygonaceae) in rodents. Arch Med Health Sci [serial online] 2015 [cited 2019 Dec 11];3:12-7. Available from: http://www.amhsjournal.org/text.asp?2015/3/1/12/154919


  Introduction Top


Herbs are plants or plant parts used for its scent, flavor, or therapeutic properties. [1] Herbal medicine remains largely an unproven, inexact science. Although the history of herbal medicine provides decades, sometimes centuries, of anecdotal information, scientific study of herbal medicine is relatively new. Despite the criticism of herbal medicine among mainstream medical professionals, it is wise to remember that many common drugs we use today were derived from plant-based sources. For example, scientists originally derived aspirin from willow bark; herbalists prescribe white willow for headaches and pain control. There are a number advantages associated with using herbal medicines as opposed to pharmaceutical products. [2]

Polygonum minus Huds. (Polygonaceae) is also known as 'laksa leaf' or 'Kesum' by the Malaysians and it is often used in cooking to add flavor to the dish since olden days. In Malaysia, several types of fresh vegetables are consumed raw, locally known as 'ulam' or equivalent to salad in other countries. These include 'selom' (Oenanthe javanica), 'pegaga' (Centella asiatica) and 'kesum'. [3] P. minus is an annual plant and distributed Britain, Scandinavia, Spain and Southeast Asian countries namely Malaysia, Thailand, Vietnam and Indonesia. [4] P. minus is an herbaceous plant which is described as shrubby, lanky and creepy. The leaves are long and lanceolate measuring about 5-7 cm long and 0.5-2.0 cm wide. The dark green leaves are aromatic and arranged alternately on the stem. The stem is cylindrical, green and slightly reddish having short internodes with nodes that are easily rooted. Generally, there are two types of kesum available namely creepy and vertical growing. P. minus leaf contains flavone, methyl flavonol and essential oil such as decanal, dodecanal, 1-decanol, 1-dodecanol, undecanal, tetradecanal, 1-undecanol, nonanal, 1-nonanol and β-caryophyllene. It is rich in beta carotene, vitamin A, vitamin C and other minerals such as potassium, calcium and phosphorus.

P. minus leaves is used for digestion, getting rid of dandruff in scalp and as tonic taken after birth. As a natural source of aliphatic aldehyde, kesum oil is believed to have high potential in flavor. [5] The antioxidant, antimicrobial and antiulcer activities of the P. minus are well-known. It promotes high levels of free radical scavenging activity and reducing power compared to other natural antioxidants. [6],[7],[8] The analgesic, anti-inflammatory and antipyretic effect of the P. minus leaves is unclear, hence the present study was planned to determine the analgesic, anti-inflammatory and antipyretic effect of the aqueous and methanol extract of P. minus in rodents.


  Materials and Methods Top


Animals

Wistar albino strain of mice, weighing about 20-30 g and Wistar rats weighing 150-200 g were obtained from Central animal House, AIMST University and used in the experiments. The experiment protocol was approved by the AIMST University Human and Animal Ethics Committee (AUHAEC 70/FOP/2011). Animals were kept in animal house at an ambient temperature of 25°C and 45-55% relative humidity, with 12 hour each of dark and light cycles. Animals were fed pellet diet and water ad-libitum. All the experiments are conducted as per AIMST University animal care guidelines which are based on the guidelines of Animal Research Review Panel.

Plant collection

The taxonomically identified Polygonum minus Huds. (Polygonaceae) plant was collected in sub-urban areas of Penang, Malaysia. The plant was identified and authenticated by pharmacogonsist in Faculty of Pharmacy, AIMST University, Bedong, Malaysia. The plant leaves were washed with running water for 10 minutes and dried under the shadow for 3 days.

Preparation of methanol extract of Polygonum minus (MEPM)

The powdered leaf of P. minus was extracted with methanol by simple maceration for 5 days. The extract was filtered, concentrated and the solid marc was weighed, percentage yield was calculated.

Preparation of aqueous extract of Polygonum minus (AEPM)

The powdered leaf of P. minus was extracted with distilled water by simple maceration for 5 days. During maceration period, few drops of chloroform were added to avoid fungal growth. The extract was filtered, concentrated and the solid marc was weighed, percentage yield was calculated.

Phytochemical analysis

Phytochemical screening test were performed for the MEPM and AEPM to check the presence of reducing sugars, cardioglycosides, anthroquinones, terpenoids, flavonoids, saponins, tannins and alkaloids by using standard procedures. [9]

Analgesic effects of AEPM and MEPM in mice

The analgesic effect of AEPM and MEPM were determined using acetic acid- induced writhing test, tail immersion method and formalin test at the dose levels of 100 and 200 mg/kg respectively. The analgesic effect of AEPM and MEPM was compared with indomethacin (250 mg/kg), a methylated- indole belongs to the acetic acid derivative class of NSAIDs.

Acetic acid- induced writhing test in mice

Either gender of Wistar albino mice were divided into six groups (five animals/group) as follows:

Group I : Normal control

Group II : Standard drug (Indomethacin 250mg/kg)

Group III : AEPM 100 mg/kg

Group IV : AEPM 200 mg/kg

Group V : MEPM 100 mg/kg

Group VI : MEPM 200 mg/kg

AEPM and MEPM were dissolved in 0.5% of CMC and administered orally. Thirty minutes after vehicle or drug administration, all the mice were given intraperitoneal injection of 0.1 ml/10 g 1.1% acetic acid and the time was immediately recorded. The mice were observed for first writhing (latency) and duration of writhing. The writhing times between the groups were compared.

Tail immersion method in mice

The animals were tested for its tail flick response, and the animals which did not showed any tail flicking response in 20 sec were excluded from the study. Pre-screened either gender of Wistar albino mice were divided into six groups (five animals/group) as follows:

Group I : Normal control

Group II : Standard drug (Indomethacin 250mg/kg)

Group III : AEPM 100 mg/kg

Group IV : AEPM 200 mg/kg

Group V : MEPM 100 mg/kg

Group VI : MEPM 200 mg/kg

The animals are housed in large spaced cages leaving the tail hanging out freely. The animals were allowed to adapt cage and experimental environment for one hour. The lower 5 cm portion of the tail was marked (immersed portion) and immersed in a cup of freshly filled hot water of 55 ± 2 °C. The reaction time was recorded in 10 sec units by a stopwatch for 20 sec. The reaction time was noted before and periodically after standard/ test drug administration (all the drugs were dissolved in 0.5% w/v of CMC and administered orally) at 1, 3 and 5 hours. [10]

Formalin test in mice

Either gender of Wistar albino mice were divided into six groups (five animals/group) as follows:

Group I : Normal control

Group II : Standard drug (Indomethacin 250mg/kg)

Group III : AEPM 100 mg/kg

Group IV : AEPM 200 mg/kg

Group V : MEPM 100 mg/kg

Group VI : MEPM 200 mg/kg

AEPM and MEPM were dissolved in 0.5% w/v of CMC and administered orally. Forty-five minutes after vehicle or drug administration, all the mice were subcutaneously administered with 0.2 ml of 3% formalin in the mouse right hind paw, and the licking time was recorded from 0-5 minutes and from 30 minutes after the formalin injection. The mean of licking time was determined and compared with the mean for the control group.

Anti-inflammatory activity of AEPM and MEPM by carrageenan induced paw edema model

Either gender of Wistar albino mice were divided into six groups (five animals/group) as follows:

Group I : Normal control

Group II : Standard drug (Indomethacin 250 mg/kg)

Group III : AEPM 100 mg/kg

Group IV : AEPM 200 mg/kg

Group V : MEPM 100 mg/kg

Group VI : MEPM 200 mg/kg

The animals were housed in large spaced cages and baseline paw volume (right hind paw) was taken. The animal was pre-treated with extract or standard drug (administered orally) 1 hour before 0.1% of carrageenan (in CMC) solution into the sub-plantar region of right hind paw. The paw volume was measured displacement of mercury in plethysmometer (Inco, India) at 0, 1, 3, 5 and 12 hours after the carrageenan administration. The inflammation reduction was compared with vehicle control group. [11]

Antiulcer activity of AEPM and MEPM by pyloric ligation model

The rats (180-200 g) were divided in to four different groups each of six animals as follows:

Group I : Normal control

Group II : Standard drug (Ranitidine 10 mg/kg)

Group III : AEPM 200 mg/kg

Group IV : MEPM 200 mg/kg

AEPM and MEPM were dissolved in 0.5% w/v of CMC and administered orally for five days. Before administering the last dose of ranitidine/AEPM/MEPM the animals were fasted for 12 hours, but they had free access to water. After one last dosing, pyloric ligation was done for all the animals as described. The animal was anesthetized with the help of anesthetic ether and the abdomen was opened by a small midline incision below the xiphoid process. The pyrloic portion of the stomach was slightly lifted out and ligated according to the method of shay et al and the stomach was replaced carefully, abdominal wall was closed by sutures. After 4 hours of pyrloic ligation, the animals were sacrificed by an over dose of anesthetic ether. The abdomen was opened and cardiac end of the stomach was dissected out for collecting the gastric juice. The volume of gastric juice was measured and centrifuged at 2500 rpm for 10 min. From the aliquot, one ml of supernatant was taken for determination of pH and total acidity. The stomach as scored according to information severity. [12],[13] On the day of pyloric ligation the animals were placed in cages with grating floor to avoid coprophagy and divided.

Determination of pH

One ml of gastric juice was diluted with one ml of distilled water and pH of the solution was estimated using pH meter.

Determination of total acidity

One ml of gastric juice was diluted with 1 ml of distilled water and two drops of phenolphthalein indicator was added to it and titrated with 0.01N NaOH until a permanent pale pink colour was observed. The total acidity was calculated using formula (Vol. of NaOH × N × 100)/0.1 and values are expressed as mEq/L

Antipyretic activity of AEPM and MEPM by Brewer's yeast induced pyrexia

Brewer's yeast induced pyrexia is a standard and well established model for screening of anti-pyretic activity of any investigational product. Brewer's yeast induces fever in 9-18 hours in rats. The rats (180-200 g) were divided in to four different groups each of six animals as follows:

Group I : Normal control

Group II : Standard drug (paracetamol 100 mg/kg)

Group III : AEPM 200 mg/kg

Group IV : MEPM 200 mg/kg

The rats were fasted for 12 hours before administering 20% Brewer's yeast and rectal temperature was recorded using digital telethermometer. Immediately after recording basal rectal temperature, Brewer's yeast (20% in normal saline) was administered intraperitoneally. The changes in rectal temperature were recorded in every 6 hours. After 24 hours of Brewer's yeast administration, the animals which showed a raise in temperature of at least 0.6°C (1°F) were included in the study. The pyrogenic rats were divided in to three groups (group II, III and IV) each of six animals. The drug was administered orally and the rectal temperature was checked at 1, 3, 5 and 24 hours of post dose. [14],[15]

Statistical analysis

The mean ± SEM values were calculated for each group. Statistical differences among the groups were determined using one-way or repeated measure ANOVA followed by Bonferroni multiple comparison test. P < 0.05 was considered to be significant.


  Results Top


The percentage yield of AEPM and MEPM was found to be 1.15% and 2.57% w/w respectively. Phytochemical analysis of the MEPM and AEPM showed the presence of reducing sugar, terpenoids, saponins, tannins, flavonoids in both extracts.

Both MEPM and AEPM significant analgesic effect in acetic acid writhing, tail immersion and formalin induced pain methods. In tail immersion method, both the extract showed marked analgesic effects, but the effect was not equivalent to that of standard. At the end of the fifth hour, MEPM 200 mg/kg showed equivalent effect when compare to the standard (delayed action). In formalin test, significant reductions in number/duration of paw licking (latency) observed at fifth and twenteith minutes. Both AEPM and MEPM showed significant reduction in latency when compare to the control [Table 1] [Table 2] [Table 3]. In all the models MEPM showed better analgesic activity than AEPM.
Table 1: Analgesic effect of aqueous and methanolic extract of P. minus in mice using acetic acid induced writhes method

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Table 2: Analgesic effect of aqueous and methanolic extract of P. minus in mice using tail immersion method

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Table 3: Analgesic effect of aqueous and methanolic extract of P. minus in mice using formalin test in mice

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The anti-inflammatory activity of AEPM and MEPM was studied using carrageenan induced paw edema model. In acute model, only AEPM showed anti-inflammatory action and this effect was comparable with that of standard drug. In carrageenan induced paw edema model, MEPM did not show any significant anti-inflammatory activity [Table 4], but MEPM showed significant anti-ulcer activity against pyloric ligated rats and AEPM failed to prevent acidic gastritis [Table 5]. In brewer's yeast induced pyrexia rat, both AEPM and MEPM did not showed any significant antipyretic effect [Table 6].
Table 4: Anti-inflammatory effect of aqueous and methanolic extract of P. minus in rats using carrageenan induced paw edema model

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Table 5: Antiulcer effect of aqueous and methanolic extract of P. minus in rats using pyloric ligation model

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Table 6: Antipyretic effect of aqueous and methanolic extract of P. minus in rats using brewer's yeast induced pyrexia

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  Discussion Top


In this study aqueous extract of P. minus showed significant analgesic and anti-inflammatory activity and methanol extract shows showed analgesic and anti-ulcer activity. This difference in activity may be due to the variations in phytoconstituents present in the extracts. Qader et al., reported the gastroprotective effect of ethyl acetate: Methanol (1:1) fraction of Polygonum minus in SD rats, and the study results showed significant gastroprotective effect against ethanol induced gastric irritation model and the gastroprotective effect was dose dependent. [6]

Herbs are the important source of drugs and many drugs are derived from herbs, including atropine, digoxin, d-tubocurarine, etc. Non-steroidal anti-inflammatory drugs are act by inhibiting by cyclooxygenase and production of prostaglandins. The presence of flavonoids is known to inhibit the prostaglandin synthetase. The chemical analysis of the MEPM, and AEPM showed the presence of flavonoids, and the pharmacological actions (anti-inflammatory/ analgesic/antipyretic effects) of the AEPM and MEPM may be due to flavonoids present in the plant. [16] Urones et al, isolated two important flavone from Polygonum minus viz., 7-methylenedioxy-5, 3´, 4´, 5´-tetramethoxyflavone and 6, 7-4´, 5´-dimethylenedioxy-3, 5, 3´-trimethoxy flavone and the pharmacological actions of isolated phytoconstituents remain unknown. [17]

Antioxidant effect of P. minus is well known. [18] Antioxidant effect of P. minus is comparable with Zingiber officinale and Curcuma longa. The total phenol content may contribute for the antioxidant effect of P. minus. The antioxidant properties may potentiate the pharmacological actions of P. minus. [19]


  Conclusion Top


The present study does confirm that, the aqueous extract of the P. minus does exhibit significant analgesic and anti-inflammatory action, whereas methanolic extract showed presence of analgesic and anti-ulcer activity. Both aqueous and methanolic extract did not show any significant antipyretic activity.

 
  References Top

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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]


This article has been cited by
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Regulatory Toxicology and Pharmacology. 2017; 86: 33
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