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www.wjpr.net Vol 3, Issue 3, 2014. 4106

ANTICONVULSANT ACTIVITY OF HYDROALCOHOLIC EXTRACT

OF OCIMUM GRATISSIMUM LEAVES

*

Sreenu Thalla1, K.Venkata Ramana1, K.Suresh Kumar, V.Lavanya Reddy1, Y.Anil Kumar2

*1

A.S.N Pharmacy College, Tenali, Guntur (dt), Andhra Pradesh, India. 2

Hindu College of Pharmacy, Amaravathi road, Guntur, Andhra Pradesh, India.

ABSTRACT

Ocimum gratissimum is used traditionally in India to treat psychoses, epilepsy, and rheumatism. The present study is aimed at investigating the anticonvulsant activity of chloroform and hydromethanolic extract of the leaves of Ocimum gratissimum which is commonly used in the Indian traditional medicine. The anticonvulsant activity of the extract was evaluated using Pentylenetetrazol (PTZ) - induced convulsions in mice and maximum electro shock (MES) -induced convulsions and lithium- pilocarpine induced status epilepticus in rats. In MES Model, both the extracts (100 and 200 mg/kg p.o.) significantly reduced the duration of hind limb extension. In the PTZ model, both extracts (100 and 200 mg/kg p.o.) delayed the latency tomyoclonic spasm and clonic convulsions significantly. In Lithium-pilocarpine model, chloroform extract (100 mg) and hydromethanolic extract (100 & 200 mg) delayed the latency to rearing with forelimb clonus significantly. The results indicate that the extracts of Ocimum gratissimum possess biologically active compound(s) that have anticonvulsant properties, which support the ethnomedicinal use of the plant as antiepileptic agents.

Keywords: Ocimum gratissimum. Lithium . pilocarpine .MES . PTZ

INTRODUCTION

Despite of advancement in modern medicine, the treatment of epilepsy is still grossly inadequate. Several plants have been described in traditional system ofmedicine in India since ancient times to treat epilepsy[1]. Traditional uses of this plant in treatment of epilepsy, insanity, jaundice, yellow fever, syphilis, helminthiasis, cough, and constipation[2]. There

Volume 3, Issue 3, 4106-4112. Research Article ISSN 2277 – 7105

Article Received on 12 January 2014, Revised on 03 February 2014,

Accepted on 23 March 2014

*Correspondence for

Author

Sreenu Thalla

A.S.N Pharmacy College,

Tenali, Guntur (dt), Andhra

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www.wjpr.net Vol 3, Issue 3, 2014. 4107 have been several efforts to verify the traditional uses of Ocimum gratissimum using the principles of modern medicine. Ocimum gratissimum possesses biologically active compound(s) that have anxiolytic and sedative properties, supporting the ethnomedicinal use of the plant as antipsychotic and antiepileptic agents. However there is no recording of anticonvulsant activity of leaves[3]. Therefore we assessed the spectrum of anticonvulsant activity of chloroformextract (CE) and hydromethanolic extract (HME) against MES, PTZ, and Lithium pilocarpine (Li-Pilo.) induced status epilepticus[4].

MATERIALS & METHODS Animals

Male Wistar rats weighing (150-200g) were obtained and they were maintained in animal house as per IAEC guidelines. Animals were access to standard pellet diet and water given ad libitum. The study was approved by Institutional animal ethical committee, IAEC/132/2013.

Plant material

The leaves of Ocimum gratissimum used in the present study was collected from the natural habitat in and around Chennai, Tamilnadu and the plant material was authentified by Dr.P.Jayaraman Ph.D., Plant Anatomy Research Centre(PARC),Tambaram. Voucher number is PARC/2012/803.

Plant extraction: The fresh leaves of Ocimum gratissimum were collected. It was defatted using petroleum ether. The marc obtained was dried and subjected to extraction by adding dried leaf powder of distilled water (1:10), heated to 50-600C under constant stirring conditions for 1hour and filtered. The methanol extract was prepared by using Soxhlet’s appataratus[5,6].

Phytochemical screening: The alcoholic extracts obtained were subjected to preliminary phytochemical screening to identify the chemical constituents. The methods of analysis employed were those described by Harbon et al., 1973.

Assessment of anticonvulsant activity

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www.wjpr.net Vol 3, Issue 3, 2014. 4108 reference standard. In case of Lithium pilocarpine induced status-epilepticus the doses of CE andHME were 100 and 200 mg/kg because the plant contains antihistaminic and anticholinergic constituents. Peripheral anticholinergic activity reduces mortality in this animal model[8].

Maximal electroshock induced seizures (MES)

The albino rats divided in groups of 6, received vehicle, CE & HME (100 or 200 mg/kg p.o.) or phenytoin sodium (40 mg/kg i.p.) 60 min before application of electric current (150 mA, 0.2 s.) using corneal electrodes. The duration of hind limb extension and number of rats exhibiting convulsions was noted in each animal. Pentylenetetrazole (PTZ) induced seizure The CE & HME were administered 60 min before the subcutaneous injection of PTZ (80 mg/kg). Mice were observed for latency tomyoclonic spasmand clonic convulsions. One group received Diazepam (2 mg/kg i.p) as a reference standard 30 min before PTZ[9]. The animals observed for onset of convulsion up to 30 min after PTZ. Lithium pilocarpine induced status-epilepticus. Status epilepticus was induced by administration of pilocarpine (30mg/kg i.p) 24 h after lithium carbonate (3 mEq/kg i.p). The effect of CE & HME (each 100 & 200 mg/kg, p.o) was studied on the rearing with forelimb clonus by administering both extracts 30 min. before injection of pilocarpine.Diazepam was used as a reference standard in a dose of 1 mg/kg i.p[10].

Statistical analysis

All the data were presented as Mean ± SEM. The data was analyzed by one way ANOVAfollowed by Dunnett’s test using the software ‘Primer of Statistics’. p <0.05 was considered significant.

RESULTS AND DISCUSSION

Table 1: Effect of chloroform extract (CE) and hydromethanolic extract (HME) of

Ocimum gratissimum on duration of hind limb extension in M.E.S induced convulsion in

rats

Treatment (dose: mg/kg) Duration of hind limb extension (Sec.)

% Survival

Vehicle (10) 12.5±0.39 66.66

HME (100) 8.4±0.6* 66.66

HME (200) 4.9±0.7* 100

CE (200) 10.6±0.5* 100

CE (400) 4.7±0.62* 100

[image:3.595.96.509.637.756.2]
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www.wjpr.net Vol 3, Issue 3, 2014. 4109 Seizure is a characteristic feature in epilepsy and rhythmic high frequency discharge of impulses by a group of neurons in the brain. Status epilepticus is characterized by repeated episodes of epilepsy without the patient having recovered from the previous attack. Several phytochemicals have been evaluated for assessment of anticonvulsant activity[11,12,13]. The observations emanated in the present study indicated that CE & HME of of Ocimum gratissimum possessed anticonvulsant activity against seizure induced by MES, PTZ, and lithium-pilocarpine. The MES test predicts activity against generalized tonic clonic and cortical focal seizures and the PTZ test against absence seizure, while Lithium-pilocarpine predicts usefulness in status epilepticus[14].

Table 2 Effect of chloroform extract (CE) and hydromethanolic extract (HME) of

Ocimum gratissimum on Pentylenetetrazol- induced convulsion in mice

Treatment (dose: mg/kg)

Latency to myoclonic spasm (Sec.)

Latency to clonic

convulsion (Sec.) % of survival Vehicle (10) 80.83±2.8 120.2±6.2 0

HME (100) 310.2±13.7* 480.7±33.2 50 HME (200) 385.33±9.44* 548.0±25.92 100

CE (200) 408.5±111.4* 683.2±187.7** 33 CE (400) 462.0±29.4* 882.5±64.24** 100

Values are mean ± SEM of 6 observations, *p <0.01, **p <0.001, Oneway ANOVA followed by Dunnett’s test

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www.wjpr.net Vol 3, Issue 3, 2014. 4110 Table 3 Effect of chloroform extract (CE) and hydromethanolic extract (HME) of

Ocimum gratissimum on Lithium-pilocarpine induced convulsion in rats

Treatment (dose: mg/kg) Latency to rearing with forelimb clonus (min) Vehicle (10) 24.2±1.6

HME (100) 46.2±2.9* HME (200) 40.4±2.2* CE (200) 46.7±3.5* CE (400) 32.7±1.8 Diazepam (2) 80.9±4.5**

Values are mean ± SEM of 6 observations NS non significant

*p <0.05 and **p <0.01 One-way ANOVA followed by Dunnett’s test

Inhibition of lithium-pilocarpine-induced convulsion may be either due to involvement of serotonergic or cholinergic mechanisms[19]. The coumarins increase brain levels of GABA inhibiting GABA transaminase enzymes thereby inhibit PTZ- and lithium-pilocarpine-induced convulsions. The MES lithium-pilocarpine-induced seizures are also inhibited by alkaloids which are abundantly present in the leaves of Ocimum gratissimum. These observations support the findings of the present study. Thus it is concluded that of Ocimum gratissimum has a potential to provide anticonvulsant principles. Saponins and coumarins may be responsible for the anticonvulsant activity. Further studies are aimed at isolating the anticonvulsant phytoconstituents of this plant and understanding their mechanism of action.

CONCLUSION

It can be concluded from the study that the anticonvulsant effects of the methanolic extract of Ocimum gratissimum may be via non-specific mechanisms. However, extensive studies are needed to evaluate the precise mechanism(s), active principles, and the safety profile of the plant as a medicinal remedy for convulsive disorders.

REFERENCES

1. Ahmed AS, Kingham AD, Geoffery AC, Norman RF (1981) Isolation of bergapten and marmesin from Balanites agyptica. Planta Med 43:92–103

2. Ambawade SD, Kasture VS, Kasture SB (2002) Anticonvulsant activity of roots and rhizomes of Glycyrrhiza Glabra. Indian J Pharmacol 34:249–253

[image:5.595.134.461.129.243.2]
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www.wjpr.net Vol 3, Issue 3, 2014. 4111 4. Kokate CK. Practical Pharmacognosy, 3rd Ed, Vallabh Prakashan, New Delhi, 1994, 07. 5. Bhutada P, Mundhada Y, Bansod K, Dixit P, Umathe S, Mundhada D (2010)

Anticonvulsant activity of berberine, an isoquinoline alkaloid in mice. Epilepsy Behavior 18:207–210.

6. Curtis and Mortiz, SOD one of the most important enzymes in the enzymatic antioxidant defence system, Ind.J.Pharmacol, 2010, 246-248.

7. Chindo BA, Anuka JA, McNeil L, Yaro AH, Adamu SS, Amos S,ConnellyWK, Lees G, Gamaniel KS (2009) Anticonvulsant properties of saponins from Ficus platyphylla stem bark. Brain Res Bull 78:276–282

8. SreenuThalla, JyothibasuTammu, Subba Reddy Thalla, Nootropic activity of Ocimum gratissimum in streptozotocin induced Amnesia, AJRC, 5(12), 2012, 1437-1439.

9. Choi SY (2005) In vitro GABA transaminase inhibitory compounds from the root of Angelica dahurica. Phytother Res 19:839–845

10.Cordano G, Terrien MA, Plonsky J, Rabanal RM, Varenne P (1978) Balanitol, a new sesquiterpene from B. roxburghii, carbon-13 NMR analysis of eudesonance sesquiterpenoids. J Indian Chem Soc 55: 1148–1151

11.Hydroxybenzaldehyde from Gastrodia elata as active in the antioxidation and GABAergic neuromodulation of the rat brain. J ethnopharmacol 73:329–333

12.Kasture VS, Kasture SB, Pal SC (1996) Anticonvulsant activity of Albizzia lebbeck leaves. Indian J Exp Biol 38:78–81

13.SreenuThalla, K.Venkata Ramana, Delhiraj N. Hepatoprotective effect of Ocimum gratissimum in the Attenuation of Myocardial Infarction Induced by Musa acuminate, IJPRB (2013), 1(5), 649-654.

14.Khandelwal KR (2008) Preliminary Phytochemical Screening, In: Practical Pharmacognosy, Nirali Prakashan, 19th ed.: p85

15.Łuszczki JJ, Andres-Mach M, Gleńsk M, Skalicka-Woźniak K (2010). Anticonvulsant effects of four linear furanocoumarins, bergapten,imperatorin, oxypeucedanin, and xanthotoxin, in the mouse maximal electroshock-induced seizure model: a comparative study. Pharmacol Reports 62:1231–1236

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www.wjpr.net Vol 3, Issue 3, 2014. 4112 17.McNamara JO (1996) Drugs effective in the treatment of the epilepsies. In: Hardman JG, Limbird JE, Molinoff PB, Ruddon RW, Gillman AG (eds) Goodman and Gillman’s the pharmacological basis of therapeutics, 9th edn. McGraw Hill, New York, pp 461–486 18.SreenuThalla, K.Venkata Ramana, JyothibasuTammu, Subba Reddy Thalla,

Cytoprotective effect of Ocimum gratissimum in the Attenuation of Myocardial Infarction Induced by Musa acuminate, AJPTR, 2(6), 2012, 724-730.

Figure

Table 1: Effect of chloroform extract (CE) and hydromethanolic extract (HME) of
Table 3 Effect of chloroform extract (CE) and hydromethanolic extract (HME) of

References

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