International Journal of Scientific & Engineering Research Volume 2, Issue 8, Auguest-2011 1
ISSN 2229-5518
Evaluation of Analgesic and Antipyretic activity of Marsilea trifolia Blanco
Mohammad Didar Khan
Abstract— The main aim of this study was to find out the Antipyretic and analgesic effects of Marsilea trifolia Blanco (Family: Marsilea- ceae) fresh leaf aqueous extract, a Bangladeshi medicinal plant. The extract exhibited marked analgesic effect by reduction of writhings inducted by acetic acid at dose of 100, 200 and 300mg/kg (p.o) in mice. Analgesic effect was also observed with the hot plate device main- tained at 55°C. Moreover, the extract produced a significant inhibition (P<0,01) in yeast induced pyrexia in rats. This study confirms tradi- tional use of Marsilea trifolia Blanco as remedy for fever.
Index Terms— Marsilea trifolia Blanco, Marsileaceae, aqueous extract, analgesic, antipyretic, writhings, hot plates test.
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n Bangladesh, Marsilea trifolia Blanco, commonly known as “Aamrul”, is traditionally used for the treatment of fever and gastro-intestinal disturbance. Several studies have reported biological effects of Marsi- lea trifolia Blanco such as antimicrobial, antioxidant and cytotoxic. In this study, I examined the effect of the fresh leaves aqueous extract of Marsilea trifolia Blanco for analgesic and antipyretic activities in experimental ani-
mals.
The plant, M. trifolia was collected from Noakhali dis- trict in the month of March, 2011 and identified by Dr. M.A. Razzaque Shah, Tissue Culture Specialist, BRAC Plant Biotechnology Laboratory, Bangladesh. The Voucher specimen is deposited at the national herbarium of Bangladesh.
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The aqueous extract was prepared according to the me- thod used in Bangladeshi traditional medicine. 20g of fresh leaves was crushed in blender in 500ml of distilled water. The extract obtained was filtered, evaporated, lyophilized and stored at 4°C until further use..
Male Swiss Albino mice (20-25g) and male Sprague Daw-
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ley rats (140-240g) were housed with free access to food and water on 12h light/12h dark cycle. The temperature was maintained at 25±2°C. The animals were deprived of food for 24h before the test, but they have free access to water.
The different concentrations of leaf aqueous extract of M. trifolia (100, 200 and 300mg/kg) was administered orally to three different groups of six mice each. The reference drug (acetylsalicylic acid 100mg/kg orally) was adminis- tered orally and used as positive control. After one hour,
0.6% acetic acid solution (10ml/kg) was injected Intrape-
ritonneally.
Nociception was evaluated 15min after acetic acid injec- tion by counting the number of abdominal constrictions. A significant reduction in the number of abdominal con- tractions (treated animals) compared to the control group (received only distilled water) was considered as anal- gesic response.
Three different groups of mice received orally 100, 200 and 300mg/kg of the extract. Acetylsalicylic acid (100mg/kg) was administered orally to positive control. Distilled water (10ml/kg) was given to control group. One hour after treatment, the animals were placed on a hot plate maintained at 55±2°C. The time taken by the mice to start licking the paw or jump out of the hot plate was considered as the reaction time. The test was carried before the treatment and at 60, 90, 120, 150 and 180 min after administration.
2.5 Antipyretic Activity
Initial rectal temperature of rats was recorded using an Ellab thermometer. Hyperthermia was induced by sub- cutaneous injection of 10ml/kg of 20% aqueous suspen-
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sion of brewer's yeast. After 18h of yeast injection, the animals showing a rise in temperature inferior to 0.7°C was discarded. Aqueous extract was given orally (300,
500 and 800mg/kg). Acetylsalicylic acid (100mg/kg oral-
ly) was used as reference drug. Control group received only distilled water (10ml/kg). The temperature was recorded at 1h, 2h, 3h and 4h after treatment.
The result of the experiments was expressed as mean±S.E.M. Statistical significance was determined by the Student's test. Values with P<0,05 were considered significant.
The present study is evaluation of new analgesic and antipyretic active principles from Bangladeshi medicinal plants. The analgesic effect with chemical and thermal stimuli has been investigated. The aqueous extract of M. trifolia (100, 200 and 300mg/kg orally) presented a po- tent antinociceptive activity in acetic acid-induced writh- ings response (Table 1). This activity was dose depen- dant, so the percentages of inhibition were respectively
65.96%; 79.52% and 9.77%. The maximum of inhibition
(89.77%) was obtained with 300mg/kg of M. trifolia ex-
tract and was comparable to that obtained by acetylsali- cylic acid 100mg/kg orally).
Groups | Dose (mg/kg ) | Number of writhings | Inhibi- tion (%) | |
Control (distilled water) | - | 55.33 ± 0,42 | - | |
Acetylsalicylic acid | 100 | 4.33 ± 0,55 | 92.17 * | |
Aqueous extract | 100 | 18.83 ± 4,02 | 65.96 * | |
200 | 11.33 ± 1,74 | 79.52 * | ||
300 | 5.66 ± 0,21 | 89.77 * | ||
Each value represents the mean ± S.E.M. (n=6) * P < 0,001 compared with control. |
This activity has been confirmed by the results of hot plate method (Table 2). However, the extract (300mg/kg) exhibited significant analgesic effect 90min after adminis- tration. Acetic acid and hot plate tests are used to study the peripheral and central analgesic effects. So the results obtained may be supported the ability of the aqueous extract to have peripheral and central pain inhibition mechanisms.
Other property exhibited by M. trifolia is antipyretic
effect.
A dose dependant antipyretic effect observed with dif-
ferent doses (300-800mg/kg orally) (Table 3). The aqueous extract showed significant degree of antipyretic activity at dose of 500 and 800mg/kg. The reduction of hyperthermia was pronounced 60 min after administra- tion and was prolonged for three hours. Subcutaneous injection of yeast induces pyrexia by synthesis of pros- taglandin. Since, the inhibition of prostaglandin may be responsible for antipyretic effect of aqueous extract of M. trifolia.
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Groups | Dose (mg/ kg) | Rectal tem- pera- ture °C before yeast injec- tion | Rectal temperature °C after yeast injection | ||||
Groups | Dose (mg/ kg) | Rectal tem- pera- ture °C before yeast injec- tion | 0h | 1h | 2h | 3h | 4h |
Control (distill- ed water) | - | 38,37± 0,09 | 39,3 3±0, 08 | 39,5 7±0, 14 | 39,5 8±0, 11 | 39,5 0±0, 12 | 39,6 2±0, 12 |
Acetyl- salicylic acid | 100 | 37,85± 0,15 | 38,9 2±0, 09 | 38,1 2±0, 07** | 37,7 8±0, 17** | 37,5 5±0, 06** | 37,9 7±0, 16** |
Aqueo us ex- tract | 300 | 37,45± 0,15 | 38,4 0±0, 21 | 38,3 8±0, 26* | 38,2 7±0, 26* | 38,0 8±0, 19** | 38,0 0±0, 16** |
500 | 37,40± 0,12 | 38,5 8±0, 16 | 38,2 6±0, 20** | 38,1 5±0, 18** | 37,8 8±0, 15** | 37,9 5±0, 20** | |
800 | 37,20± 0,11 | 38,7 0±0, 12 | 37,7 2±0, 33** | 37,4 0±0, 35** | 37,3 3±0, 25** | 37,6 5±0, 34** | |
Each value represents the mean ± S.E.M. (n=6) ** P < 0,001 compared with control. *P < 0,01 |
The present study indicated that the aqueous extract of the aerial part of M. trifolia have got profound analgesic and antipyretic effect and may have potential use in medicine.
In comparison with the positive control (acetylsalicylic acid), the analgesic and antipyretic activities exhibited by the aqueous extract of the plant showed potent activity. This clearly indicates the presence of potent bioactive principles in this aqueous extract which might be very useful as analgesic and antipyretic agent.
Further studies comprising of phytochemical investiga-
tions of the used plant and evaluation for analgesic and
antipyretic activities using other methods (e.g. various biochemical assays both in vivo and in vitro) are essential to characterize them. It may be concluded from this study that M. trifolia is active in the tested animals. In addition, the results confirm the use of the plant in tradi- tional medicine. The results of the investigation do not reveal that which chemical compound is responsible for aforementioned activity. Now my next aim is to explore the lead compound liable for aforementioned activity from this plant.
[1]. Cox, P.A. and M.J. Balick, 1994. “The ethnobotanical approach
to drug discovery”, Scientific American 270, pp. 60-65.
[2]. Dubey, N.K., R. Kumar and P. Tripathi, 2004. “Global promotion of herbal medicines: India’s opportunity”. Current Science 86, pp. 37-41.
[3]. Duke. J. A. and E. S. Ayensu, 1985. “Medicinal Plants of China”,
Reference Publications Inc. ISBN 0-917256-20-4.
[4]. Fransworth N.R., 1993. “Ethnopharmacology and future drug development: the North American experience”, Journal of Ethnopharmacol 38, pp. 45-152.
[5]. Houghton, P.J., 1995. “The role of plants in traditional medicine and current therapy”, Journal of Altern and Complement Med 1, pp. 131-143.
[6]. Sadhu, S. K.; E. Okuyama, H. Fujimoto and M. Ishibashi,
2003."Separation of Leucas aspera, a medicinal plant of Bangladesh,
guided by prostaglandin inhibitory and antioxidant activities" Chemical & Pharmaceutical Bulletin 51, pp.595-598.
[7]. Schofield, J. J., 1989. “Discovering Wild plants, Alaska, Western Canada, the Northwest”. Alaska Northwest Books, G.TE Discovery Publications, Inc. 22023 20th Ave. S.E. Bothell, WA. 98021.
[8]. Kapoor, L. D., Srivastava, S. N., Singh, A., Kapoor, S. L. & Shah, N. C. (1972). Survey of indian plants of saponins alkaloids and flavonoïdes. Liodya, 35: 288-295.
[9]. Bentley, G. A., Newton, s. H.&Starr, J. (1983). Studies on the antinociceptive action of agonist drugs and their interactions with opioïds mechanism. British Pharmacol., 79: 125-134.
[10]. Abena, A., Diatewa, M., Gakosso, G., Gbeassor, M. & Hondi Assah, J. M. (2003). Analgesic, antipyretic and ant-inflammatory effects of essential oil of Lippia Multiflora. Fitoter., 74: 231-236.
[11]. Markouk, M., Lazrek, H. B.&Jana, M. (1999).Analgesic effect of extracts from Cotula cinerea L. Phyto. Res., 13: 229-230.
[12]. Koster, R.,Anderson, M.&De Beer, E. J. (1959).Acetic acid for analgesic screening. Fed., Proc., 18: 412-412.
[13]. Al-Ghamdy, M. S. (2001). Theantiiflammatory, analgesic and antipyretic activity of Nigella sativa. J. pharmacol., 76: 45-48.
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