|Year : 2016 | Volume
| Issue : 1 | Page : 25-28
Absence of anticonvulsant activity in Asparagus adscendens Roxb. hydroethanolic root extract against acute pentylenetetrazol and maximal electroshock-induced convulsion mice models
Priyanka Pahwa, Rajesh Kumar Goel
Department of Pharmaceutical Sciences and Drug Research, Punjabi University, Patiala, Punjab, India
|Date of Web Publication||19-Feb-2016|
Rajesh Kumar Goel
Department of Pharmaceutical Sciences and Drug Research, Pharmacology Division, Punjabi University, Patiala - 147 002, Punjab
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Introduction: The use of Asparagus adscendens (family:Liliaceae) root powder has been reported traditionally for the treatment of epilepsy. But, it is yet to be validated pharmacologically. Therefore, the present study was undertaken to explore the anticonvulsant effect of the roots in the experimental animal models of convulsions. Materials and Methods: The anticonvulsant effect of hydroethanolic root extract of A. adscendens (AAE) was studied at 25, 50, and 100 mg/kg; intraperitoneally (i.p.) in maximal electroshock (MES), and at 25, 50, 100, and 200 mg/kg; i.p. doses in pentylenetetrazol (PTZ) test in mice. The duration of tonic hind limb extension (s) and latency to tonic-clonic convulsions (min) was noted in MES and PTZ tests, respectively. Phenytoin (25 mg/kg; i.p.) and diazepam (5 mg/kg; i.p.) served as reference standards in MES and PTZ tests, respectively. Percentage mortality was also noted. Results: The AAE treatment did not show any protective effect with regard to induction and duration of tonic hind limb extensor in MES test and latency to tonic-clonic convulsions in PTZ test, as compared to their respective controls. Conclusions: The results obtained from the experiments indicate that the AAE lacks anticonvulsant activity in MES- and PTZ-induced convulsion tests.
Keywords: Asparagus adscendens Roxb., convulsions, epilepsy, maximal electroshock, pentylenetetrazol
|How to cite this article:|
Pahwa P, Goel RK. Absence of anticonvulsant activity in Asparagus adscendens Roxb. hydroethanolic root extract against acute pentylenetetrazol and maximal electroshock-induced convulsion mice models. J Pharm Negative Results 2016;7:25-8
|How to cite this URL:|
Pahwa P, Goel RK. Absence of anticonvulsant activity in Asparagus adscendens Roxb. hydroethanolic root extract against acute pentylenetetrazol and maximal electroshock-induced convulsion mice models. J Pharm Negative Results [serial online] 2016 [cited 2018 May 23];7:25-8. Available from: http://www.pnrjournal.com/text.asp?2016/7/1/25/177057
| Introduction|| |
Epilepsy is a chronic neurological disorder, characterized by the episodic and unpredictable occurrence of epileptic seizures, which affects around 50 million people worldwide.  Despite the availability of several antiepileptic drugs, many drugs limits seizures spread to some extent but a large number of patients suffer from their side effects.  Thus, nowadays researchers diverted their area of interests toward medicinal plants as a source of complementary and alternative medicine. 
In this regard, literature reports have suggested that the root powder of Asparagus adscendens has been traditionally claimed to be effective in the management of epilepsy.  Apart from the traditional use in epilepsy, A. adscendens has also been used for the treatment of diarrhea, dysentery and general debility, galactagogue, demulcent, and general tonic.  A. adscendens have also been reported to possess several other pharmacological activities, such as anti-filarial,  insulin enhancing activity,  in vitro AChE and BChE inhibition activity,  chemo-modulatory potential, antioxidant activity,  and in sexual disorders.  The phytochemical research carried out on its roots had led to the isolation of spirostanol glycosides (asparanin C and D), furostanol glycosides (asparoside C and D), , and shatavarin IV  as a main phytoconstituents. A. racemosus, another species form this genus, was found to be phytochemically, morphologically, and microscopically similar to A. adscendens. Hence, there might be chances of alternative/substitute use of both the species.  Moreover, A. racemosus has already been reported for its antiepileptogenic  and anticonvulsant activity. , Therefore, A. adscendens might have an anticonvulsant activity which has not been reported yet. Hence, an effort was made to pharmacologically validate the anticonvulsant potential of hydroethanolic root extract of A. adscendens (AAE) was evaluated using maximal electroshock (MES)- and pentylenetetrazol (PTZ)-induced convulsions.
| Materials And Methods|| |
Plant material, preparation, and dosing of crude extracts
The roots of A. adscendens Roxb. were collected from the Council of Scientific and Industrial Research (CSIR) - Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh in June 2013 and authentication was done by Dr. Bikram Singh, CSIR- Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh (voucher no: PLP16565).
The authenticated plant parts were cleaned, shade-dried, and grounded into a moderately coarse powder. The powdered material (100 g) was subjected to extraction by 50% ethanol + 50% distilled water using percolation at room temperature. The filtered material was evaporated under reduced pressure using rotavapor (Buchi Type Rotary Vacuum Evaporator, Axiva, Shanghai, China). The remaining viscous liquid was subjected to freeze-drying using lyophilizer (Delvac, India) to get powered hydroethanolic root extract.
Based on the results of some pilot studies carried out on a limited number of animals, three doses were selected for MES test (25, 50, and 100 mg/kg), and four doses for PTZ test (25, 50, 100, and 200 mg/kg). Dosing was carried out by dissolving the extract in normal saline and was injected intraperitoneally (i.p.). The vehicle control groups were injected with equal volume of vehicle (normal saline) i.p. In all cases, the injection volume was kept 10 ml/kg.
Preliminary phytochemical screening
Preliminary phytochemical screening of the crude extract was carried out qualitatively for the presence of alkaloids, glycosides, flavonoids, saponins, sterols, tannins, phenolic compounds, amino acids, proteins, fatty acids, carbohydrates, and terpenes by using different phytochemical screening tests. 
Swiss albino mice weighing 20-30 g were purchased from Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana. The animals were housed on a 12 h light/dark cycle under controlled temperature (22°C ± 2°C) and humidity (50% ±10%). They were allowed to acclimate for 1 week with ad libitum access to food and water. The procedures in this study were conducted in accordance to the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals, ministry of environment and forest, Government of India, and approved by the Institutional Animal Ethical Committee.
Drugs and chemicals
PTZ was procured from Sigma-Aldrich, Co., St. Louis, MO, USA. Phenytoin sodium and diazepam were obtained locally from Jackson Laboratories Pvt. Ltd., India. All the drugs were dissolved in normal saline.
Acute toxicity test of the extract
The A. adscendens extract was administered at different doses, up to 10 times of effective dose, i.e. 25, 50, 100, 200, and 400 mg/kg; i.p., to six different groups of mice (n = 6). One group served as control and received an equal volume of vehicle. , Percentage mortality and gross behavioral changes were observed during 24 h after treatment.
Pentylenetetrazol-induced convulsions test
The PTZ convulsions test was performed by the method of Swinyard et al. with slight modifications. , Mice in different groups were injected with the varying doses of AAE (25, 50, 100 and 200 mg/kg), vehicle, and diazepam (5 mg/kg). After 30 min of these treatments, all groups were injected with PTZ 75 mg/kg. Latency to tonic- clonic convulsions (min) and percentage protection were noted and compared with that of vehicle control group.
Maximal electroshock-induced convulsions test
MES convulsions in mice were induced by delivering a calibrated (through a current calibrator [Rolex, Ambala, India]) trans-auricular electroshock of 56 mA for 0.2 s using a convulsiometer (Rolex, Ambala, India) via a pair of crocodile ear clips.  Mice in different groups were injected with the varying doses of AAE (25, 50 and 100 mg/kg), vehicle, and phenytoin (25 mg/kg). After 30 min of these treatments, all groups were subjected to the MES-induced convulsions. Duration of tonic hind limb extension(s) (THLEs) was noted and compared with the vehicle control group.
All the results were expressed as a mean ± standard error of mean. Data were analyzed using one-way analysis of variance followed by Dunnett's test. The results were regarded as significant at P < 0.001.
| Results|| |
Preliminary phytochemical screening
The extraction of root powder yielded 44.7% w/w of crude hydroethanolic extract. The results of preliminary phytochemical screening confirmed the presence of carbohydrates and glycosides, flavonoid, saponins, steroids, triterpenoids, and amino acids in the extract.
Acute toxicity studies
In acute toxicity test, no mortality was observed in animals treated with the extract at all doses. There was no change in behavior of animals during 24 h of administration of the extract. The extract was found to have no neurotoxic effects up to highest dose.
Effect on pentylenetetrazol-induced convulsions
In PTZ-induced convulsions tests, treatment with AAE showed a slight dose-dependent increase in the latency to tonic-clonic convulsions, however, the change observed in comparison to vehicle control group was not significant [Table 1]. There was a decrease in percentage mortality at lower doses of the extract. Diazepam-treated group showed a significantly (P < 0.001) increase in latency to tonic-clonic convulsions as compared to vehicle control group.
|Table 1: Effect of the hydroethanolic root extract of Asparagus adscendens on pentylenetetrazol-induced convulsions |
Click here to view
Effect of maximal electroshock-induced convulsions
In MES-induced convulsions, treatment with the AAE failed to decrease the duration of the THLE in a dose-dependent manner [Table 2]. There was no significant effect observed on the percentage of animals showing THLE. However, phenytoin-treated group showed significantly (P < 0.001) decrease in duration to THLE as compared to vehicle control group.
|Table 2: Effect of the hydroethanolic root extract of Asparagus adscendens on maximal electroshock-induced convulsions |
Click here to view
| Discussion|| |
The present study was envisaged to validate the traditionally claimed anticonvulsant potential of A. adscendens roots. The phytochemical investigation of crude extract (yielded 44.7% w/w) revealed the presence of carbohydrates and glycosides, flavonoid, saponins, steroids, triterpenoids, and amino acids. Moreover, the AAE at a dose range of 25, 50, 100, and 200 mg/kg; i.p., had no effect on the general behavior of animals.
To screen the new antiepileptic drugs, there is a diversity of animal models that could potentially be used, but PTZ and MES-induced convulsion models were served as "gold standards" tests in the early stages of drug testing.  Clinically, the active anticonvulsants have been found to be protective in at least one of the above tests.  Therefore, both of above models were employed in the present study. In PTZ-induced convulsions test, the AAE treatment showed the slightly dose-dependent increase in the latency to tonic-clonic convulsions (but insignificant) up to 200 mg/kg while the MES - induced convulsion test was also found to be ineffective as no decrease in duration in THLE was observed at any dose. In line to this, the AAE neither acts through GABAergic neurotransmission nor through voltage-gated sodium channel. Similarly, on the basis of clinical evidence various active anticonvulsant drugs such as levetiracetam, seletracetam, etc., are ineffective against acute PTZ- and MES-induced convulsion models, but active in other models. , Hence, the AAE might be effective in other models such as 6-Hz model and kindling models.
| Conclusion|| |
The results of the current study demonstrate the insignificant anticonvulsant activity of AAE against PTZ- and MES-induced convulsion models, indicating its ineffectiveness against generalized tonic-clonic convulsions. Thus, further research is warranted to support the traditional antiepileptic use of the A. adscendens roots.
The authors are deeply grateful to the Council of Scientific and Industrial Research (CSIR), Pusa, New Delhi, India for providing financial assistance (Vide F. No. 38[(339)/12/EMR-II) for the project, and project fellowship to Miss. Priyanka Pahwa. We are also thankful to Dr. Bikram Singh, CSIR - Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh for the authentication of plant material.
Financial support and sponsorship
Council of Scientific and Industrial Research (CSIR), Pusa, New Delhi, India for providing financial assistance (Vide F. No. 38(1339)/12/ EMR-II) for the project, and project fellowship to Miss. Priyanka Pahwa.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Pahwa P,Goel RK. Ameliorative effect of Asparagus racemosus root extract against pentylenetetrazol-induced kindling and associated depression and memory deficit. Epilepsy Behav 2016. [doi: 10.1016/j.yebeh.2016.01.025] (In Press).
Sahu S, Dutta G, Mandal N, Goswami AR, Ghosh T. Anticonvulsant effect of Marsilia quadrifolia
Linn. on pentylenetetrazole induced seizure: A behavioral and EEG study in rats. J Ethnopharmacol 2012;141:537-41.
Abdollahi Fard M, Shojaii A. Efficacy of Iranian traditional medicine in the treatment of epilepsy. Biomed Res Int 2013;2013:692751.
Sharma S, Kumar A. Herbal medicinal plants of Rajasthan: Musli. Int J Life Sci Pharm Res 2012;2:L62-8.
Khan I, Nisar M, Khan N, Saeed M, Nadeem S, Fazal-ur-Rehman, et al
. Structural insights to investigate Conypododiol as a dual cholinesterase inhibitor from Asparagus adscendens
. Fitoterapia 2010;81:1020-5.
Singh R, Khan NV, Singhal KC. Potential antifilarial activity of roots of Asparagus adscendens
Roxb, against setaria cervi in vitro
. Indian J Exp Biol 1997;35:168-72.
Mathews JN, Flatt PR, Abdel-Wahab YH. Asparagus adscendens
(Shweta musli) stimulates insulin secretion, insulin action and inhibits starch digestion. Br J Nutr 2006;95:576-81.
Singh M, Singh S, Kale RK. Chemomodulatory potential of Asparagus adscendens
against murine skin and forestomach papillomagenesis. Eur J Cancer Prev 2011;20:240-7.
Bansode FW, Arya KR, Singh RK, Narender T. Dose-dependent effects of Asparagus adscendens
root (AARR) extract on the anabolic, reproductive and sexual behavioural activity in rats. Pharm Biol 2015;53:192-200.
Sharma SC, Chand R, Sati OP. Steroidal saponins of Asparagus adscendens
Roots. Phytochemistry 1982;21:2075-8.
Bhutani KK, Jadhav AN. Steroidal saponins from the roots of Asparagus adscendens
roxb and Asparagus racemosus
willd. Indian J Chem 2006;45B: 1515-24.
Sharma OP, Kumar N, Singh B, Bhat TK. An Improved method for thin layer chromatographic analysis of saponins. Food Chem 2012;132:671-4.
Panda SK, Das D, Tripathy NK. Botanical studies of plants sold in market as safed musli. Int J Pharm Res Develop 2011;2:42-51.
Amberkar MV, Christopher R, Chandrashekar BR, Pradeepa HD, Kumari M. An experimental evaluation of anticonvulsant activity of Asparagus racemosus
. Int J Pharm Sci Res 2011;11:64-8.
Jalapure SS, Bagewadi V, Shaikh I. Antiepileptic effect of Asparagus racemosus
root extracts. J Trop Med Plants 2009;10:157-61.
Trease G, Evans SM. Pharmacognosy. 15 th
ed. London: Bailer Tindal; 2002. p. 193-393 .
Singh D, Goel RK. Anticonvulsant effect of Ficus religiosa: Role of serotonergic pathways. J Ethnopharmacol 2009;123:330-4.
Sonavane GS, Palekar RC, Kasture VS, Kasture SB. Anticonvulsant and behavioural actions of Myristica fragrans
seeds. Indian J Pharmacol 2002;34:332-8.
Kaur M, Goel RK. Anti-convulsant activity of Boerhaavia diffusa
: Plausible role of calcium channel antagonism. Evid Based Complement Alternat Med 2011;2011:310420.
Swinyard EA, Brown WC, Goodman LS. Comparative assays of antiepileptic drugs in mice and rats. J Pharmacol Exp Ther 1952;106:319-30.
Singh D, Singh B, Goel RK. Hydroethanolic leaf extract of Ficus religiosa
lacks anticonvulsant activity in acute electro and chemo convulsion mice models. J Pharmaceut Neg Results 2011;2:58-61.
Rogawaski MA, Porter RJ. Antiepileptic drugs: Pharmacological mechanisms and clinical efficiency with consideration of promising developmental stage compounds. Pharmacol Rev 1990;42:233-86.
Malawska B. New anticonvulsant agents. Curr Top Med Chem 2005;5:69-85.
Rogawski MA. Diverse mechanisms of antiepileptic drugs in the development pipeline. Epilepsy Res 2006;69:273-94.
Klitgaard H, Matagne A, Gobert J, Wülfert E. Evidence for a unique profile of levetiracetam in rodent models of seizures and epilepsy. Eur J Pharmacol 1998;353:191-206.
[Table 1], [Table 2]