|Year : 2016 | Volume
| Issue : 1 | Page : 33-36
Insignificant anticonvulsant activity of Padina tetrastromatica (Brown macroalgae) in mice
Subhash R Yende1, Uday N Harle2, Abhay M Ittadwar1
1 Department of Pharmacology, Gurunanak College of Pharmacy, Nagpur, Maharashtra, India
2 Clinical Research Consultant, Nagpur, Maharashtra, India
|Date of Web Publication||19-Feb-2016|
Subhash R Yende
Department of Pharmacology, Gurunanak College of Pharmacy, Nagpur, Maharashtra
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Introduction: Marine macroalgae or seaweeds are found in the coastal region have created a promising significance in the biomedical area, mainly because of their contents of bioactive substances. The objective of the present study was to investigate the anticonvulsant activity of chloroform and ethanol extracts of Padina tetrastromatica (PT), a marine macroalgae (brown algae) in mice. Materials and Methods: The anticonvulsant activity of chloroform and ethanol extracts of PT was studied at 400 and 600 mg/kg, against maximal electroshock (MES) and pentylenetetrazole (PTZ) induced convulsion in mice. The duration of tonic hind limb extension (THLE), latency to onset of clonic convulsions and percent protection was noted in MES and PTZ tests, respectively. Phenytoin (25 mg/kg) and phenobarbitone (20 mg/kg) served as reference standards. Results: The chloroform extract of PT at 600 mg/kg significantly decreased the duration of THLE, while ethanol extract did not alter the duration of THLE in MES model. Further, chloroform and ethanol extracts of PT was found to be ineffective as an anticonvulsant when assessed by PTZ-induced convulsive model, as compared to their respective vehicle-treated groups. Conclusion: From the results of the present study it can be concluded that the chloroform extract of PT at 600 mg/kg showed significant anticonvulsant activity, while other extracts lack anticonvulsant activity in MES and PTZ model. However, further studies are required using different animal models to support these findings.
Keywords: Anticonvulsant activity, brown algae, maximal electroshock-induced convulsion, Padina tetrastromatica, pentylenetetrazole-induced convulsion
|How to cite this article:|
Yende SR, Harle UN, Ittadwar AM. Insignificant anticonvulsant activity of Padina tetrastromatica (Brown macroalgae) in mice. J Pharm Negative Results 2016;7:33-6
|How to cite this URL:|
Yende SR, Harle UN, Ittadwar AM. Insignificant anticonvulsant activity of Padina tetrastromatica (Brown macroalgae) in mice. J Pharm Negative Results [serial online] 2016 [cited 2018 May 23];7:33-6. Available from: http://www.pnrjournal.com/text.asp?2016/7/1/33/177061
| Introduction|| |
Epilepsy is a chronic neurological disorder characterized by recurrent derangement of the nervous system due to sudden excessive disorderly discharge from the cerebral neurons.  There are a number of synthetic anticonvulsant drugs currently available for management, control, and/or treatment of epilepsy. However, most of these synthetic drugs are associated with serious side effects including teratogenicity, chronic toxicity, and adverse effects on cognition and behavior. , Therefore, there is a dire need for the development of cheap, effective, and safe anticonvulsant agents from plants and other natural sources.
Marine macroalgae or seaweeds are found in the coastal region between high tide to low tide and in the subtidal region up to a depth where 0.01% photosynthetic light is available and can be classified into three classes; brown algae (Phaeophyta), green algae (Chlorophyta), and red algae (Rhodophyta). Marine macroalgae have created a promising significance in the biomedical area, mainly because of their contents of bioactive substances. Polysaccharides, terpenoids, phlorotannins, fucoidans, sterols, and glycolipids obtained from marine macroalgae showed a wide range of pharmacological properties which includes anticancer, anti-inflammatory, antimicrobial, antiviral, antioxidant, hypoglycemic, hepatoprotective, and neuroprotective activities. ,, Also, some marine organism and marine macroalgae showed the potential as a source of new drugs for the treatment of neurological disorders. , Many traditional herbs and herbal medicines have been reported for their central nervous system (CNS) activities, whereas the marine flora has not explored up to that extent. Hence, we undertook the study to evaluate CNS potential of some marine macroalgae.
Padina tetrastromatica (PT) (Hauck) is a brown alga found in the coastal areas of India. Methanolic extract of PT have reported for spasmogenic, antifertility, hypotensive properties,  and in vitro antioxidant activity. , Sulphated polysaccharide from PT showed significant anti-inflammatory activity against carrageenan-induced paw edema.  Moreover, it reported to possess antihepatitis B virus activity.  Furthermore, chloroform, ethanol, and water extract of PT showed antimicrobial activity against Gram-positive, Gram-negative bacterial, and fungal test.  However, anticonvulsant activity of PT has not been investigated. Hence, we evaluated the anticonvulsant activity of chloroform and ethanol extracts of PT against seizure induced by maximal electroshock (MES) and pentylenetetrazole (PTZ) in mice.
| Materials And Methods|| |
Swiss albino mice (25-30 g) were used for the experiment purpose. The animals were housed in solid-bottomed polypropylene cages and acclimatized to animal house conditions. The mice were fed with commercial standard diet and water ad libitum. The experiments were designed and conducted in accordance with the guideline of the CPCSEA, Government of India and approved by the Institutional Animal Ethical Committee (Approval No. GNCP/IAEC/2011-12/P'cology-01).
Drugs and chemicals
PTZ (Sigma-Aldrich, USA), phenytoin (Zydus Cadila, Ahmedabad, India), and phenobarbitone (Abbott India Ltd.,) were used in this study. The drug was diluted with distilled water before use.
Seaweed collection and extracts preparation
The brown seaweeds, PT were collected from the inter-tidal rocky shore of Bhatkarwada, Ratnagiri coast in November-December 2011. The seaweed species was identified by Professor B. B. Chaughule, Emeritus Professor, Department of Botany, University of Pune, Pune (India). The fresh samples were washed with sea water followed by fresh water to remove salts, epiphytes, microorganisms, and other suspended materials, and dried at room temperature. The air-dried and coarsely powdered sample was extracted successively using Soxhlet apparatus by Petroleum Ether, Chloroform and Ethanol.
The chloroform and ethanol extracts of PT were analyzed for the presence of phytochemicals by qualitative analysis. ,
Acute toxicity studies
The acute toxicity was determined on Swiss albino mice as per the OECD-423 guidelines.  The overnight-fasted animals were administered extracts orally at the dose level of 2000 and 5000 mg/kg body weight and were continuously observed for 2 h to detect changes in the autonomic or behavioral responses and then, monitored for any mortality for the following 7 days.
Maximal electroshock-induced convulsion
Mice were divided into six groups consisting of five mice in each group. Group I served as vehicle treated group (5% Tween-80 p.o.), Group II as a standard drug pretreatment group (Phenytoin 25 mg/kg, i.p.), Group III and IV treated with chloroform extract of PT (CPT 400 and 600 mg/kg p.o.), and Group V and VI treated with ethanol extract of PT (EPT 400 and 600 mg/kg p.o.), respectively. Group I, III-VI animals received extract treatment continuously for 7 days, and electroshock was applied after 1 h of the last dose of respective treatments, whereas Group II animals received phenytoin (25 mg/kg, i.p.) 30 min prior to the electroshock. Seizure was induced using electroconvulsiometer (INCO, Ambala, India). An electric shock (45 mA, 0.2 s) was applied to each mouse through ear-clip electrode, and duration of tonic hind limb extension (THLE) and percent protection was recorded. 
Mice were divided into six groups consisting of five mice in each group. Group I served as vehicle treated group (5% Tween-80 p.o.), Group II as a standard drug pretreatment group (Phenobarbitone 20 mg/kg, i.p.), Group III and IV treated with (CPT 400 and 600 mg/kg p.o.), and Group V and VI treated with (EPT 400 and 600 mg/kg p.o.), respectively. All mice received PTZ (80 mg/kg, i.p.) 1 h after their respective treatment, except Group II that received PTZ after 30 min and observed for the onset of clonic convulsion, duration of THLE, and percent protection. ,
Results were expressed as a mean ± standard error of mean. The data were analyzed using one-way analysis of variance followed by Dunnett's test, and P < 0.05 was considered as statistically significant.
| Results|| |
The qualitative phytochemical analysis revealed the presence of steroids, terpenoids, flavonoids, alkaloids and glycosides in chloroform extract and steroids, alkaloids, and glycoside in EPT.
Acute toxicity studies
Since no mortality was observed at 2000 mg/kg as well as at 5000 mg/kg, doses of 100, 200, and 400 mg/kg of PT extracts were selected. But, 100 and 200 mg/kg of PT extracts were showed an insignificant effect in the preliminary behavioral test. Therefore, 400 and 600 mg/kg of PT extracts were selected for further study.
Maximal electroshock-induced convulsion
In MES test, phenytoin (25 mg/kg) and CPT (600 mg/kg) showed significant (P < 0.05) decrease in the duration of THLE. However, CPT (400 mg/kg) and EPT (400 and 600 mg/kg) showed an insignificant effect on the duration of THLE as compared with the vehicle-treated group [Table 1].
|Table 1: Effect of Padina tetrastromatica on tonic hind limb extension and percent protection on maximal electroshock-induced convulsion |
Click here to view
Treatment with phenobarbitone (20 mg/kg) significantly increased (P < 0.01) the latency to onset of clonic convulsion as well as the duration of THLE. While, CPT (400 and 600 mg/kg) and EPT (400 and 600 mg/kg) did not significantly increase the latency to onset of clonic convulsion and duration of THLE, as compared with the vehicle-treated group. Further, extract administration showed 20% protection against PTZ-induced seizure [Table 2].
|Table 2: Effect of Padina tetrastromatica on onset of clonic convulsion, tonic hind limb extension and percent protection on pentylenetetrazole-induced convulsion |
Click here to view
| Discussion|| |
The MES test is considered to be a predictor of likely therapeutic efficacy against generalized tonic-clonic seizures, this tonic extension can be prevented either by a drug that inhibits voltage-dependent Na + channels, such as phenytoin, valproate, and lamotrigine.  The PTZ-induced convulsion test is a valid model for human generalized myoclonic and absence seizures. PTZ may be exerting its convulsive effect by inhibiting the activity of gamma aminobutyric acid (GABA) at GABA A receptors, the major inhibitory neurotransmitter which is implicated in epilepsy. It has been indicated that PTZ-induced seizures can be prevented by drugs that reduce T-type Ca 2+ currents, such as ethosuximide, and also by drugs that enhance GABA A receptor-mediated inhibitory neurotransmissions, such as benzodiazepines and phenobarbital. ,
The finding of this study showed that CPT at 600 mg/kg significantly decreased the duration of THLE, while ethanol extract did not alter the duration of THLE in MES model. Further, chloroform and ethanol extracts of PT were found to be ineffective as an anticonvulsant when assessed by PTZ-induced convulsive model.
| Conclusion|| |
From this, it can be concluded that the CPT at 600 mg/kg can be effective against generalized tonic-clonic seizures and may act by inhibiting voltage-dependent Na + channels. While ethanol extract was ineffective against MES model. Further, both chloroform and ethanol extracts lack anticonvulsant activity in PTZ model. Hence, further studies are required from other regions using other models of convulsion to support or negate the anticonvulsant effects of PT.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Bhosle V. Anticonvulsant and antioxidant activity of aqueous leaves extract of Desmodium triflorum
in mice against pentylenetetrazole and maximal electroshock induced convulsion. Rev Bras Farmacogn 2013;23:692-8.
Tripathi KD, editor. Essentials of Medical Pharmacology. 6 th
ed. New Delhi, India: Jaypee Brothers Medical Publishers (P) Ltd.; 2008. p. 401-5.
Arzimanoglou A, Hirsch E, Nehlig A, Castelnau P, Gressens P, Pereira de Vasconcelos A. Epilepsy and neuroprotection: An illustrated review. Epileptic Disord 2002;4:173-82.
Smit AJ. Medicinal and pharmaceutical uses of seaweed natural products: A review. J Applied Phycol 2004;16:245-62.
Liu L, Heinrich M, Myers S, Dworjanyn SA. Towards a better understanding of medicinal uses of the brown seaweed Sargassum
in traditional Chinese medicine: A phytochemical and pharmacological review. J Ethnopharmacol 2012;142:591-619.
Yende SR, Harle UN, Chaugule BB. Therapeutic potential and health benefits of Sargassum
species. Pharmacogn Rev 2014:815;1-7.
Nunez R, Garateix A, Laguna A, Fernández MD, Ortiz E, Llanio M, et al
. Caribbean marine biodiversity as a source of new compounds of biomedical interest and others industrial applications. Pharmacologyonline 2006;3:111-9.
Pangestuti R, Kim SK. Neuroprotective effects of marine algae. Mar Drugs 2011;95:803-18.
Naqvi SW, Kamat SY, Fernandes L, Reddy CV, Bhakuni DS, Dhawan BN. Screening of some marine plants from the Indian coast for biological activity. Bot Mar 1980;24:51-6.
Chandini SK, Ganesan P, Bhaskar N. In vitro
antioxidant activities of three selected brown seaweeds of India. Food Chem 2008;107:707-13.
Chew YL, Lim YY, Omar M, Khoo KS. Antioxidant activity of three edible seaweeds from two areas in South East Asia. LWT Food Sci Technol 2008;41:1067-72.
Mohsin S, Kurup GM. Mechanism underlying the anti-inflammatory effect of sulphated polysaccharide from Padina tetrastromatica
against carrageenan induced paw edema in rats. Biomed Prev Nutr 2011;1:294-301.
Subramaniam D, Nawabjan N, Malayan J, Mohanam L, Vaikuntam M, Manickan E. Anti hepatitis B virus (HBV) activity of marine brown algae, Padina tetrastromatica.
Vet Sci Res 2011;2;25-9.
Rangaiah SG, Lakshmi P, Manjula E. Antimicrobial activity of seaweeds Gracillaria, Padina
and Sargassum sps
on clinical and Phytopathogens. Int J Chem Anal Sci 2010;1;114-7.
Trease GE, Evans WC. Textbook of Pharmacognosy. 12 th
ed. London: Bailere Tindall; 1983. p. 169-357.
Jana S, Shekhawat GS. Phytochemical analysis and antibacterial screening of in vivo
and in vitro
extracts of Indian medicinal herb: Anethum graveolens
. Res J Med Plant 2010;4:206-12.
Acute Oral Toxicity: OECD guidelines for testing of chemicals- 423. (AOT); Environmental Health and Safety Monograph Series on Testing and Assessment. 2001. p. 1-14.
Swinyard EA, Brown WC, Goodman LS. Comparative assays of antiepileptic drugs in mice and rats. J Pharmacol Exp Ther 1952;106:319-30.
Jain NN, Ohal CC, Shroff SK, Bhutada RH, Somani RS, Kasture VS, et al
. Clitoria ternatea
and the CNS. Pharmacol Biochem Behav 2003;75:529-36.
Luszczki JJ, Antkiewicz-Michaluk L, Czuczwar SJ. 1-Methyl-1,2,3, 4-tetrahydroisoquinoline enhances the anticonvulsant action of carbamazepine and valproate in the mouse maximal electroshock seizure model. Neuropharmacology 2006;50:133-42.
McNamara JO. Pharmacotherapy of the epilepsies. In: Bruton LL, Lazo JS, Parker KL, editors. Goodman and Gilman's The Pharmacological Basis of Therapeutics. 11 th
ed. New York: McGraw Hill; 2006. p. 501.
Meldrum BS. GABA agonists as antiepileptic agents. Neurology 1994;44:514-23.
[Table 1], [Table 2]