ORIGINAL ARTICLE
Year : 2013 | Volume
: 1 | Issue : 1 | Page : 6--9
Anti-convulsant activity of ethanolic extract of Moringa concanensis leaves in Swiss albino mice
Anu Elizabeth Joy1, Shankar Bhat Kunhikatta1, Shyamjith Manikkoth2,
1 Department of Physiology, Yenepoya Medical college, Mangalore, Karnataka, India
2 Department of Pharmacology, Yenepoya Medical college, Mangalore, Karnataka, India
Correspondence Address:
Shyamjith Manikkoth
Department of Pharmacology, Yenepoya Medical College, Mangalore - 575 018, Karnataka
India
Abstract
Objectives : To investigate the anti-convulsant activity of ethanolic extract of Moringa concanensis leaves on maximal electroshock-induced seizures (MES) and pentylenetetrazole (PTZ)-induced seizures in Swiss albino mice. Materials and Methods : The ethanolic extract of Moringa concanensis leaves (200 mg/kg, i.p) was studied for its anti-convulsant effect on MES and PTZ-induced seizures in Swiss albino mice. In MES seizures, suppression of tonic hind limb extension was noted. In PTZ seizures, abolition of the convulsions was noted. Results : The ethanolic extract of the leaves of Moringa concanensis (200 mg/kg, i.p) significantly (P < 0.001) abolished the hind limb extension induced by MES. The same dose also significantly (P < 0.001) protected the animals from PTZ-induced tonic convulsions. Conclusion : The data suggests that the ethanolic extract of Moringa concanensis leaves may produce its anti-convulsant effects via multiple mechanisms since it abolished the hind limb extension induced by MES as well as abolished seizures produced by PTZ.
How to cite this article:
Joy AE, Kunhikatta SB, Manikkoth S. Anti-convulsant activity of ethanolic extract of Moringa concanensis leaves in Swiss albino mice.Arch Med Health Sci 2013;1:6-9
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How to cite this URL:
Joy AE, Kunhikatta SB, Manikkoth S. Anti-convulsant activity of ethanolic extract of Moringa concanensis leaves in Swiss albino mice. Arch Med Health Sci [serial online] 2013 [cited 2023 Mar 23 ];1:6-9
Available from: https://www.amhsjournal.org/text.asp?2013/1/1/6/113548 |
Full Text
Introduction
Epilepsy is a CNS disorder associated with recurrent episodes of seizures due to the abnormal electrical activity in the brain. Nearly 40 million people all over the world are affected by this disease. [1] In India, overall prevalence rate of this neurological disorder is 5.59 per 1000 population. [2] Currently, many drugs are available for treating this disorder, but use of these anti-epileptic agents are associated with many adverse effects. Moreover, these agents fail to provide a complete cure. Hence, there is a need for a potent anti-epileptic agent, which is devoid of side-effects. Plants and their phytoconstituents have important role in the development of a potent anti-convulsant agent. [3],[4]
Moringa concanensis belongs to the family Moringaceae. [5] It is abundantly seen in Perambalur district of Tamilnadu [commonly known as Kattu murungai or Peyi murungai] [6] and in Warangal district of Andhra Pradesh [commonly known as Adavi Mulaga, Konda mulga]. [7],[8] This small tree with strong central trunk, thick bark, bipinnate leaves and with an intense horseradish odor has vast therapeutic uses in traditional medicine. [9] Around 20 types of human ailments can be cured by using different parts of this plant. [6] Seed oil can be used in sunscreen. [10] Its analgesic and anti-inflammatory, [11] anti-pyretic properties [12]were proved in pre-clinical studies.
Materials and Methods
Drugs and chemicals
Phenytoin sodium (Abbott Group, Acme formulation Pvt. Ltd., Himachal Pradesh), Sodium Valproate (Sun Pharma, Sikkim) was obtained from a pharmacy in Mangalore. Pentylene tetrazole (Sigma Aldrich, China) was obtained from Rajesh Chemicals, Mumbai.
Instruments
Soxhlet apparatus was used for preparing the plant extract.
Electro-convulsiometer (INCO) was used for inducing convulsions.
Plant material
The leaves were used for the study. Moringa concanensis leaves were collected from Warangal district of Andhra Pradesh. They were authenticated by Dr. Noeline J. Pinto, Head of Botany department, St. Agnes College, Mangalore, Karnataka, India. The leaves were shade-dried and then grinded into coarse powder.
Preparation of the Extracts
Moringa concanensis ethanolic extract (MCEE): A weighed quantity (500 g) of the coarse powder was taken and extracted with ethanol (90%) in a Soxhlet apparatus. The extract was concentrated on a water bath at a temperature not exceeding 60°C (yield 10% w/w). The ethanolic extract was suspended in distilled water. MCEE was administered at a dose of 200 mg/kg/day i.p.
Animals
Swiss albino mice of either sex weighing 25-30 gms were used in this study after obtaining Institutional Animal Ethical Committee Clearance (IAEC), Yenepoya University. The mice were maintained under standard conditions in the Animal House (CPCSEA approved, Reg. No: 347) under Dept of Pharmacology, Yenepoya University, Mangalore. The mice were kept in Polypropylene cages (U. N. Shah manufacturers, Mumbai) under standard housing conditions and maintained on standard pellet diet (Amrut Lab Animal Feed, Pranav Agro Industries Ltd., Sangli, Maharashtra) and water ad libitum. The mice were maintained on a 12:12 hour light-dark cycle.
Acute Toxicity Study [13]
Acute toxicity study was performed according to the OECD guidelines on Swiss albino mice, and the animals were kept overnight fasting providing water ad libitum, after which the extracts were administered orally 1600 mg/kg and observed for the mortality.
Anti-Convulsant Activity [13]
a. Maximal electroshock (MES) seizure
Electrical stimulation was applied using ear electrodes. The electrodes were moistened with saline before application. All animals were stimulated with 50 mA for 0.2 seconds, with constant voltage stimulators of 250 V. [14] The animals were divided into 3 groups. Each group consisting of 6 males and 6 females (n = 12).
Group I: Normal Saline (0.1 ml i.p for 10 days) + MES on 10 th day
Group II: MCEE (200 mg/kg body weight i.p for 10 days) + MES on 10 th day
Group III: Phenytoin (25 mg/kg body weight i.p for 10 days) + MES on 10 th day
On 10 th day, the test samples were given 1 hour prior to induction of convulsions. Suppression of tonic hind limb extension was taken as a measure of efficacy in this test.
b. Pentylene tetrazole (PTZ)-induced convulsion
PTZ 70 mg/kg i.p was administered to mice. The parameter noted was duration of convulsions. [14] The animals were divided into 3 groups. Each group consisting of 6 males and 6 females (n = 12).
Group I: Normal Saline (0.1 ml i.p for 10 days) + PTZ on 10 th day
Group II: MCEE (200 mg/kg body weight i.p for 10 days) + PTZ on 10 th day
Group III: Sodium Valproate (75 mg/kg body weight i.p for 10 days) + PTZ on 10 th day
On 10 th day, the test samples were given 1 hour prior to induction of convulsions. Abolition of the convulsions was taken as a measure of efficacy in this test.
Phytochemical Screening of Mcee
A preliminary phytochemical analysis of the ethanolic extract was carried out for the presence of various phytoconstituents like alkaloids, saponins, tannins, phenols, flavanoids, and carbohydrates. [15]
Analysis
Statistical Significance
The results of the study is expressed as Mean ± SD, n = 12. One Way ANOVA was used to analyze and compare the data, followed by Tukey Krammer multiple comparison tests.
Results
Acute toxicity study
There was no mortality among the graded dose groups of animals, and they did not show any toxicity or behavioral changes at a dose level of 1600 mg/kg. This finding suggests that MCEE was safe and non-toxic to rats up to 1600 mg/kg. Hence, a lower dose of 200 mg/kg was selected for this study.
Anti-epileptic activity
MES-induced seizures
In the case of MES-induced seizures, there was no hind limb extension in MCEE and Phenytoin groups on comparing with the control group (P < 0.001). There were 4 deaths in the control group. No mortality was observed in MCEE and Phenytoin groups [Table 1].{Table 1}PTZ-induced seizuresIn the case of PTZ-induced seizures, there was considerably significant decrease (P < 0.001) in the mean duration of convulsions in MCEE and Sodium Valproate groups on comparing with the control group. There were 9 deaths in the control group. No mortality was observed in MCEE and Sodium Valproate groups [Table 2].{Table 2}
Phytochemical screening of the MCEE
Preliminary phytochemical analysis of the ethanolic extract confirmed the presence of various phytoconstituents like alkaloids, tannins, phenols, flavanoids, and carbohydrates [Table 3].{Table 3}
Discussion
Among the test used for screening the anti-convulsant activity of an agent, MES and PTZ test are regarded as the best. MES seizures resembles generalized tonic and clonic seizures in humans and PTZ-induced convulsions resembles the absent seizures in humans. [16]
The drugs that block the voltage-gated sodium channel and NMDA receptor abolishes MES seizures. The drugs that block T-type calcium channels in the thalamus or the agents which has GABA mimicking activity will prevent PTZ seizures. [17]
Moreover, MES- and PTZ-induced convulsions are associated with oxidative damage. [18],[19]
Since Moringa concanensis abolishes both MES and PTZ seizures, it might posses sodium channel blockade, NMDA blockade, calcium channel blockade, or GABA agonist activity. The anti-convulsant activity of Moringa concanensis can also be due to the antioxidant property. The anti-convulsant activity can be due to the presence of various phytoconstituents like alkaloids, tannins, phenols, flavanoids, and carbohydrates. Further studies are ongoing using different extracts of Moringa concanensis to elucidate the exact mechanism by which this plant acts as an anti-epileptic agent.
Acknowledgement
I hereby thank Dr. Shankar Bhat K, Professor and Head, Department of Physiology, Dr. S N Rao, Professor and Head, Department of Pharmacology, Yenepoya Medical College, Mangalore for their valuable advice and guidance.
References
| 1 | Ngo Bum E, Taiwe GS, Moto FC, Ngoupaye GT, Vougat RR, Sakoue VD, et al. Antiepileptic medicinal plants used in traditional medicine to treat epilepsy, clinical and genetic aspects of epilepsy. In: Zaid Afawi 2011. ISBN: 978-953-307-700-0. Available from: http://www.intechopen.com/books/clinical-and-genetic-aspects-ofepilepsy/antiepileptic-medicinal-plants-used-in-traditional-medicine-to-treat-epilepsy. Croatia. [Last accessed date on 2013 Jan 13]. |
| 2 | Kumar D, Singh J, Baghotia A, Kumar S. Anticonvulsant effect of the ethanol extract of Caesalpinia pulcherrima (L.) Sw., Fabaceae, leaves. Available from: http://www.scielo.br/pdf/rbfar/2010nahead/aop1410.pdf. [Last accessed date on 2013 Jan 13]. |
| 3 | Hegde K, Thakker SP, Joshi AB, Shastry CS, Chandrasekhar KS. Anticonvulsant activity of Carissa carandas Linn. Root extract in experimental mice. Tropical J Pharm Res 2009;8:117-25. |
| 4 | Available from: http://www.cdc.gov/nccdphp/publications/aag/pdf/epilepsy.pdf. [Last accessed date on 2013 Jan 12]. |
| 5 | Paliwal R, Sharma V, Pracheta. A review on horse radish tree (Moringa Oleifera): A multipurpose tree with high economic and commercial importance. Asian J Biotechnol 2011;3:317-28. |
| 6 | Anbazhakan S, Dhandapani R, Anandhakumar P, Balu S. Traditional medicinal knowledge on Moringa concanensis Nimmo of Perambalur District, Tamilnadu. Anc Sci Life 2007;26:42-5. |
| 7 | Krishna IS, Sujatha M. Statu s s urvey of medicinal plant diversity at Kondapalli reserve forest, Andhra Pradesh, India. Int J Appl Sci Eng Technol 2012;1:1-5. |
| 8 | Reddy KN, Trimurthulu G, Reddy CS. Plants used by the ethenic people of Krishna district of Andhra Pradesh. Indian J Tradit Knowl 2010;9:313-7. |
| 9 | Manzoor M, Anwar F, Iqbal T, Bhanger MI. Physico-Chemical Characterization of Moringa concanensis Seeds and Seed Oil. J Amer Oil Chem Soc 2007;84:413-9. |
| 10 | Kale S, Gajbhiye G, Chaudhari N. Formulation and in vitro Evaluation of Moringa concanensis, Nimmo. Seed Oils Sunscreen Cream. Int J Pharm Tech Res 2010;2:2060-2. |
| 11 | Rao CV, Hussain T, Verma AR, Kumar MN. Vijayakumar GD. Reddy evaluation of the analgesic and anti-inflammatory activity of Moringa concanensis tender fruits. Asian J Tradit Med 2008;3:95-103. |
| 12 | Jayabharathi M, Chitra M. Evaluation of anti-inflammatory, analgesic and antipyretic activity of Moringa concanensis Nimmo. J Chem Pharm Res 2011;3:802-6. |
| 13 | Manikkoth S, Deepa B, Joy AE, Rao SN. Anticonvulsant activity of phyllanthus amarus in experimental animal models. Int J Appl Biol Pharm Technol 2011;2:144-9. |
| 14 | Vogel GH, Vogel WH. "Drug Discovery and Evaluation; Pharmacological assays". Germany: Verlag, Heidelberg; 1997. p. 487,696. |
| 15 | Koruthu DP, Manivarnan NK, Gopinath A, Abraham R. Antibacterial evaluation, reducing power assay and phytochemical screening of Moringa Oleifera leaf extracts: Effect of solvent polarity". Int J Pharm Sci Res 2011;2:2991-5 |
| 16 | Gopalakrishna HN, Sudhakar P, Shilin G, Ashok KS, Holla GK, Nair V, et al. Effect of Acorus calamus on electrical and chemical induced seizures in mice. Int J Appl Biol Pharm Technol 2010;1:465-72. |
| 17 | Suresh Babu AR, Karki SS Anticonvulsant activity of various extracts of leaves of Calotropis giganeta Linn against seizure induced models. Int J Pharm Pharm Sci 2011;3:200-3. |
| 18 | Rauca C, Zerbe R, Jantze H. Formation of free hydroxyl radicals after pentylenetetrazole-induced seizure and kindling. Brain Res 1999;847:347-51. |
| 19 | Rola R, Swiader M, Czuczwar SJ. Electroconvulsions elevate the levels of lipid peroxidation process in mice. Pol J Pharmacol 2002;54:521-4. |
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