Screening of Aerial Parts of the Plant Clerodendrum paniculatum Linn for Anti-Convulsant Activity

 

Priyanka K*, Dr. Kuppast I. J, Gururaj S. V, Chethan I. A

Department of Pharmacology, Sri Adichunchanagiri College of Pharmacy, Adichunchanagiri University,

B.G. Nagara, Mandya (Dist) – 571448, Karnataka.

*Corresponding Author E-mail: Priyasmg2016@gmail.com

 

ABSTRACT:

The present study was undertaken for investigating and validating the traditional claims of Clerodendrum paniculatum Linn for its anti-convulsant activity. Clerodendrum paniculatumlinn belongs to verbenacae family. Ethno medical importance of various species of Clerodendrum genus has been reported in various indigenous systems of medicines and folk medicines. It is used as a medicine for the treatment of sore eyes, urinary tract, gonorrhoea and kidney problems. However pharmacological properties of plants have not yet been extensively studied, it is therefore thought worthwhile to study about the unrevealed properties of this plant. The aerial parts of plant are used for preparing extraction using ethyl acetate, petroleum ether and aqueous as a solvent. Shade dried, powdered (40 mesh size) leaves of Clerodendrum paniculatum Linn were extracted with ethyl acetate, petroleum ether and macerated with distilled water. On using solvent extraction phytochemical screening was analyzed, it reveals the presence of flavanoids, tannins, glycosides, saponins, and terpenoids like compounds. Anti-convulsant activity of ethyl acetate extract shows significant activity comparable with standard drug, Phyntoin. Petroleum ether and aqueous extracts shows little anti-convulsant activity.

 

KEYWORDS: Clerodendrum paniculatum Linn Anti-convulsant activity.

 

 


INTRODUCTION:

Clerodendrum paniculatum Linn. (Family Verbenacea) commonly known as ‘Red Pagoda plant’ is a semi woody shrub of 1-2 m height growing naturally in shady places throughout India. It is used traditionally in India, China and Japan, in the treatment of rheumatism, neuralgia, ulcer, inflammation, and for healing wounds [1-3]. Preliminary phytochemical screening showed the presence of terpenes, flavonoids, tannins, alkaloids, phenolic acid, sterols, glycosides, phenolic acid, sterols, and glycosides.

 

 

The plant has got immense medical importance which is used for treatment for inflammation ulcer, vitratedvata, pitta wound and skin disease. A decoction of root is used as tonic for aches and pains. It is used as a medicine for the treatment of sore eyes, urinary tract, gonorrhoea and kidney problems (4,5)

 

The objective of this work is to evaluate the phytochemical analysis and anti-convulsant properties of ethyl acetate, petroleum ether and aqueous extract of aerial parts of Clerodendrum paniculatum Linn as a part of exploration of new and novel bio-active compounds.

 

MATERIALS AND METHODS:

Plant Materials:

Clerodendrum paniculatum Linn plant was collected from the local areas of Shivamogga and it has been authenticated by Dr. Rudrappa, Dean and Botanist, Department of Biological sciences, S.R.N.M. College, Shivamogga.

 

The aerial parts of Clerodendrum paniculatum Linn were shade dried and reduced to acoarse powder in a Pulverizer (Sunbeam, Munger, India) using Mesh No. 3 and passed through a Sieve No. 40 to obtain about 2 kg of powder for further analysis.

 

Preparation of extracts:

Various extracts of the plant material were prepared by soxhlet extraction method. The powdered material of Clerodendrum paniculatum Linn was extracted with different solvents (ethyl acetate, petroleum ether and aqueous,) in a soxhlet extractor for 48 hrs in 8 batches of 50g each. The extract was concentrated in vacuum using rotary flash evaporator (Buchi, Flawil, Switzerland). The solvent was removed completely over the water bath and finally desiccator dried. The extract, so obtained was labelled, weighed and the yield was calculated in terms of grams percent of the weight of the powdered aerial parts of the plant. These extracts are then used for the activities.

 

Preliminary screening:

The presence of bio- active compounds was screened for saponins, flavanoids, terpenoids, saponins, glycosides and tannins. (6,7)

 

Animals:

Healthy young adult male and non-pregnant female Swiss albino mice (20 – 30g) and rats (150 – 200 g) of Wistar strain of either sex were used for the acute toxicity and pharmacological studies (anti-convulsant) using ethyl acetate, petroleum ether and aqueous extracts of the aerial parts of the plant Clerodendrumpaniculatum linn. The animals were procured from Central Animal House, National College of Pharmacy, Shivamogga, Karnataka. After randomization into various groups, animals were acclimatized for period of 10 days under standard husbandry conditions. Room temperature 270 ± 300C, Relative humidity 65 ± 10%, 12 hours– Light/dark cycle

 

All the animals were fed with rodent pellet diet (Gold Mohr, Lipton India Ltd.,) and water was allowed ad-libitum under strict hygienic condition. Ethical Clearance (NCP/IAEC/CL/04/2016-17) for performing experiments on animals was obtained from Institutional Animal Ethical Committee (IAEC)

 

Statistical analysis:

All the values were expressed as mean ± S.E.M. Statistical analysis was carried out by performing one-way ANOVA followed by Pair wise comparisons of Tukey'sHSD (honestly significant difference) test. A probability level of P<0.05 was considered significant, P<0.01 is considered as moderately significant and P<0.001 is considered as highly significant.

 

Acute toxicity study8:

Acute oral toxicity study for the proprietary formulation was carried out using OECDguideline 425 (modified, adopted 23 rd march 2006). Tween-80 (1%) was used as a vehicle to suspend the extracts and was administered orally and the first animal receives a dose step below the level of the best estimate of the LD50 and dose progression factor should be chosen to be the antilog of 1/(the estimated slope of the dose-response curve) and should remain constant throughout testing (a progression of 3.2 corresponds to a slope of 4). The testing samples were prepared by suspending the different extracts (ethyl acetate, petroleum ether and aqueous) in distilled water using tween 80 (1%) as suspending agent. The initial dose in this experiment was 200mg/kg body weight and there was no mortality or toxicity in animals hence this dose is considered as lethal. To study pharmacological activities the fraction was administered in the dose of 200mg/kg body weight which is equal to 1/10th of 2000mg/kg body weight.

 

 Ethyl acetate, petroleum ether and aqueous extracts of Clerodendrumpaniculatumlinnwere used to investigate the following anti-convulsant activity at the dose of 200mg/kg body weight.

 

Anti-convulsant activity:

Convulsions are also known as seizures and epilepsy. Anti-convulsant are the drugs used to abolish the seizures. Anti-epileptic are the drugs which are administered prophylactically to prevent the seizures. Anti-convulsant activity was performed by maximal electro-shock induced convulsions and PTZ induced convulsion. Phenytoin was taken as standard reference drug. Two models have been used to evaluate the anti-convulsant activity,

 

A. Maximal Electric Shock Induced convulsion

(In-vivo)9,10:

In this model, the animals were divided into five groups with six animals in each group. The animals of group I served as solvent control, received distilled water (1 ml/100gm b.w.); group II receive phenytoin (4 mg/kg b.w), treated as positive control; III, IV and V groups treated with total petroleum ether and ethyl acetate extract of the plant at the dose of 200mg/kg and 200mg/kg b.w. respectively. All the treated groups were administered orally1hour prior to the electric shock induced in the entire animals passing a current of 50 mA for 0.2 sec duration through electro convlsiometer (Techno India) using ear electrodes. The duration of flexion, extensor, clonus and stupor phase were noted and percentage ofinhibition of seizures relative to controls wascalculated.

B. Pentylenetetrazole – induced seizure model9,10:

In this type of seizure model, the animals were divided into five groups with six animals in each group. Group I served as solvent control, received distilled water (1ml/100gm b.w), Group II received phenytoin (4 mg/kg b.w), treated as positive control and III, IV and V groups treated with total petroleum ether, aqueous and ethyl acetate leaves extract at the dose of 200 mg/kg b.w. respectively. All the treated groups were administered 60min prior to the administration of Pentylenetetrazole (80 mg/kg b.w) by i.p route. The animals were observed for 1 hour by placing in a separate cage. The duration of seizures (tonic, clonic convulsions) were recorded and Percentage of inhibition of seizures relative to controls wascalculated.


 

 

RESULTS AND DISCUSSION:

Anti-convulsant activity of various extracts:

 

Table No. 1: Effect of total ethyl acetate, petroleum ether and aqueous extract of aerial parts of the plant Clerodendrum paniculatum Linn on Maximal Electric Shock induced convulsion in mice after a hour.

Groups

Drug Used

Flexion (in sec)

Extension (in sec)

Clonus (in sec)

Stupor (in sec)

Recovery / Death

I

Control

8.2±0.4778

18.5±0.5187

22.25±0.4730

50.58±3.009

52.9±4.636

II

Phenytoin (80mg/kg)

1.17±0.2021

0.4±0.083

8.36±0.3903

90.56±3.496

100.5±2.73***

III

Ethyl acetate (200mg/kg)

1.75±0.2075**

10.48±0.4831*

14.01±0.3972*

75.85±3.427**

98.68±2.800***

IV

Pet ether, (200 mg/kg)

1.98±0.26*

10.28 ±0.4831**

13.59±0.5722**

70.41±3.976*

95.32±3.322

V

Aqueous, (200mg/kg)

1.82±0.2472*

11.36±0.6004NS

13.93±0.6102*

72.41±27655*

94.53±3.89

Note: Data was analysed using one way ANOVA followed by pairwise comparision. Values are expressed as mean ± S.E.M. n=6, ***P < 0.001(highly significant), **P< 0.01(moderately significant) and *P < 0.05(significant).

 

 

Figure No.1: Histogram showing the effect of Clerodendrum paniculatum Linn extracts on Convulsion after a hour of drug administration in Maximal Electric Shock Induced convulsion.

 

 

 

Table No.2. Effect of ethyl acetate, petroleum ether and aqueous extract of aerial parts of plant Clerodendrum paniculatum Linn on Pentylenetetrazole (PTZ) induced convulsion in mice after a hour.

Group

Drug used

On set of time in seconds (Mean ± SEM)

Jerks

Clonus

Extensor

I

Control

85.25±2.86

60.55±3.474

279.6±6.474

II

Phenytoin (80 mg/kg)

0.216±0.06009

0

0

III

Ethyl acetate extract (200 mg/ kg)

74.71±4.4330**

62. 05±3.602*

251.64±6.684**

IV

Petroleum ether extract (200 mg/ kg)

68.76±4.022*

65.81±2.169*

258.75±4.818

V

Aqueous extract (200 mg/ kg)

69.78±3.972ns

67.38±4.607*

254.76±3.38*

 

 

Figure No.2: Histogram showing the effect of Clerodendrum paniculatum Linn extracts on Convulsion after a hour of drug administration in Pentylenetetrazole (PTZ) Induced convulsion.

 

 

 


CONCLUSION:

Anti-convulsant activity study of ethyl acetate at the test concentrations shows significant activity (P<0.001) comparable with standard drug, Pheyntoin. Petroleum ether and aqueous extracts shows little anti-convulsant activity.

 

Thus the study implicates that the aerial parts of Clerodendrum paniculatum Linn possess significant dose dependant anti-convulsant activity and therefore provides scientific base for its use in folklore remedies as an anti-convulsant drug of natural origin. Hence there is need for further study to rationalize the active chemical entity.

 

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Received on 13.10.2018       Modified on 30.10.2018

Accepted on 16.11.2018       ©A&V Publications All right reserved

Res.  J. Pharmacology and Pharmacodynamics.2019; 11(1):01-04.

DOI: 10.5958/2321-5836.2019.00001.6