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Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 113  

Original Article 

Exploring the protective effects of aqueous extracts of Ruta Chalepensis 

Linn. On drug induced seizures in animal models 

Ramdas Bhat1, Hemalatha C H2, A R Shabaraya3 

From, 1Assistant Professor, 2PG Scholar, Department of Pharmacology, 3Principle and Head of the Department of 

Pharmaceutics, Srinivas College of Pharmacy, Valachil, Post Farangipete, Mangalore, Karnataka, India - 574143. 

ABSTRACT 

Background and Objective: Convulsion is a sudden, involuntary muscle contraction causing rapid and repetitive movements 

of the body. Ruta Chalepensis Linn. In is a medicinal plant used to treat a wide range of disorders including seizure. But, the 

anticonvulsant activity of this plant has not been studied in deep. We therefore sought to evaluate the anticonvulsant activity of 

an aqueous leaf extract of Ruta Chalepensis Linn. On convulsing induced in mice. Method: The anticonvulsant activity of an 

aqueous leaf extract of Ruta Chalepensis Linn. On convulsing induced in mice, by strychnine nitrate (2.0 mg/kg) and 

Pentylenetetrazole (PTZ) (80 mg/kg) in mice. Chronic study was conducted using low, medium and high doses of ARC (250, 

500 and 750mg/kg respectively). The above-mentioned doses were administered daily once for a period of 7 consecutive days. 

Aqueous extract (250, 500 and 750 mg/kg), diazepam (10mg/kg intraperitoneally or oral.), and distilled water (10 ml/kg, i.p) 

were administered before induction of seizures. Results: In strychnine induced convulsion test the onset of seizure was 

significantly delayed in the animals treated with Ruta Chalepensis Linn leaf aqueous extract at 750mg/kg. In PTZ induced 

convulsion model the onset of seizure was significantly delayed in the animals treated with Ruta Chalepensis Linn leaf 

aqueous extract at a dose of 750mg/kg. Conclusion: As a result of foregoing Ruta Chalepensis Linn leaf aqueous extract has 

considerable diuretic activity at higher doses. 

Key words: Aqueous extracts of Ruta chalepensis Linn (AERC), Strychnine-induced convulsion test, Pentylenetetrazole (PTZ) 

induced Seizures, Epilepsy, Glycine, GABAnergic. 

pilepsy is a neurological disorder characterized by 

recurrent and unpredictable seizures caused by 

sudden bursts of abnormal electrical activity in the 

brain. Seizures can be of different types, including tonic-

clonic, myoclonic, absence, and others, and are often 

treated with anticonvulsant medications to reduce the risk 

of seizures [1]. Despite the availability of different medi-

cations, there are still a considerable number of patients 

who experience inadequate seizure control or suffer from 

medication side effects. This has led to a search for 

alternative treatment options, including herbal remedies 

that have been used for centuries to treat seizures [2]. Ruta 

chalepensis Linn. Is a medicinal plant that has been 

traditionally used for various ailments, including seizures. 

Access this article online 

Received – 05th April 2023 

Initial Review – 08th April 2023 

Accepted – 25th April 2023 Quick Response Code 

The plant contains several bioactive compounds, such 

as alkaloids, flavonoids, and coumarins, which have 

various pharmacological activities, including anticon-

vulsant, anti-inflammatory, and analgesic effects [3]. 

Pentylenetetrazole (PTZ), strychnine are commonly used 

compounds to induce convulsions in animal models [4]. 

Strychnine is a potent convulsant that acts by blocking the 

inhibitory neurotransmitter glycine in the spinal cord, 

leading to increased excitability and ultimately, 

convulsions [5]. While PTZ-induced convulsions are 

mainly mediated by the inhibition of the neurotransmitter 

GABA, strychnine -induced convulsions are not mediated 

by GABA [6], making them a useful model for investi-

gating alternative pathways and mechanisms involved in 

seizures. The use of animal models in the study of 

convulsions and seizures has been widely accepted and 

employed in the field of epilepsy research [7]. 

_______________________________________ 

Correspondence to: Ramdas Bhat, Assistant Professor, 

Department of Pharmacology, Srinivas College of 

Pharmacy, Valachil, Post Farangipete, Mangalore, 

Karnataka, India-574143. Email: ramdas21@gmail.com 

E 

mailto:ramdas21@gmail.com


Bhat R et al.                              Protective Effects of Ruta Chalepensis Linn. Extracts on drug-induced seizures 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 114  

Animal models allow for the investigation of various 

factors that contribute to the development of convulsions, 

such as age, sex, species, diet, water, day/light cycle, 

temperature, preparation dose, and route of administration. 

Furthermore, animal models allow for the evaluation of the 

efficacy and safety of potential anticonvulsant treatments 

[8, 9]. The aqueous extracts of Ruta chalepensis Linn. 

Have been reported to have several pharmacological 

activities, including anticonvulsant effects, which makes 

them a promising candidate for further investigation. The 

evaluation of the potential protective effects of Ruta 

chalepensis Linn. Against both PTZ and strychnine 

induced convulsions in animal models may provide new 

insights into the mechanisms of action involved in these 

seizures and identify new therapeutic strategies for the 

treatment of epilepsy. 

METHODOLOGY 

Animals: The following study used mice of both sexes 

weighing around 18-22g. The animals were fed a standard 

pelleted diet (Lipton India Ltd., Mumbai) and distilled 

water ad libitum under a constant 12 hours light and dark 

cycle. Prior to the experiment, all animals were housed in 

laboratory conditions for 5 days. The animals are now 

divided into 5 groups containing 6 animals each. All 

experiments were carried out in accordance with the 

ethical standards. 

Plant extract: Plant was collected in Mangalore and 

authentication was done by Botanist, Pilikula 

Nisargadhama, Vamanjoor. The leaves are separated shade 

dried for one day and the aqueous drug extract was 

prepared by steam distillation. The extract was suspended 

in 1% tween 80, and was administered orally to the 

animals by gastric intubation using force feeding needle. 

Dose was selected as 250mg/kg and 500mg/kg and 

750mg/kg as lower, intermediate and higher dose 

respectively based on the acute toxicity study (LD50). 

Study type: Screening of Ruta chalepensis Linn leaf 

aqueous extract for its anticonvulsant activity using 

experimental animals. 

Screening models used:  

a) Strychnine-induced Convulsions Test 

b) Pentylenetetrazole (PTZ) -induced Seizures. 

Study site: Department of Pharmacology, Srinivas 

College of pharmacy, Valachil, Mangalore. 

Sample size: 60 mice were selected for the entire study 

Study Duration: 7 days 

Inclusion criteria: Mice of both sex having weight of 18-

22g was included in study  

Exclusion criteria: Mice above 25g was excluded from 

the study. 

Experimental Design 

Acute toxicity studies: Acute toxicity study was 

performed in accordance with OECD guidelines. No 

adverse effect or mortality was detected in mice up to 2 

gm/kg, p.o of Ruta chalepensis Linn during the 24 to 72 

hrs observation periods. The rats were continually 

monitored for 5 hours during this time for any obvious 

behavioral, neurological, or autonomic toxic effects, 

mortality after 24 to 72 hrs (Table 1 and 2). 

Strychnine induced convulsion: The mice utilized for 

this study were groups of six of either sex with a weight of 

between 18-22 respectively. They were now treated with 

Diazepam 10 mg/kg which is a standard drug. One hour 

after the administration of the standard drug the mice were 

injected with 2 mg/kg Strychnine nitrate i.p the time until 

occurrence of tonic extensor convulsions and death noted 

during a 1 h period. With this dose of strychnine 

convulsions were observed in 80% of the controls. The 

animals employed are mice of either sex that weigh 

between 18 and 22 g. The animals are divided into 6 

groups and 6 groups receive either the test substance or the 

reference medication through injection, intravenously, or 

orally. As a control, six mice from another group are used. 

Thirty minutes following an intravenous injection, sixty 

minutes following a subcutaneous injection PTZ is 

subcutaneously given at an 80 mg/kg dose. Each animal is 

housed in its own plastic cage for a one-hour observation 

period. There are records of seizures and tonic-clonic 

convulsions. Convulsions must be present in at least 80% 

of the animals in the control group. 

Table 1: Grouping of animals in strychnine induced convulsion 

Group Treatment Dose No. of  animals Parameters to be assessed 

Group I (control) Saline + Strychnine 10ml/kg + 2mg/kg 6  

Percentage inhibition of 

seizures relative to 

control 

Group II (standard) Diazepam + Strychnine 10mg/kg + 2mg/kg  6 

Group III (Low Dose) AERC + Strychnine 250mg/kg + 2mg/kg 6 

Group IV (Moderate Dose) AERC + Strychnine 500mg/kg + 2mg/kg 6 

Group V (High Dose) AERC + Strychnine 750mg/kg + 2mg/kg 6 



Bhat R et al.                              Protective Effects of Ruta Chalepensis Linn. Extracts on drug-induced seizures 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 115  

Table 2 - Pentylenetetrazole-induced Seizures (PTZ) 

 

Statistical analysis: All the data were expressed in mean 

± SEM. The significance of differences in mean between 

control and treated animals for different parameters 

determined by one way ANOVA followed by Dunnett’s 

multiple comparison test. Significance for difference 

between groups were evaluated for student’s t-test to come 

to final conclusion. 

RESULT AND DISCUSSION 

 

AERC leaves were screened for anticonvulsant activity 

using Strychnine and PTZ induced convulsion model in 

mice. Chronic study was conducted using low, medium 

and high doses of ARC (250,500& 750mg/kg 

respectively). The above-mentioned doses were 

administered daily once for a period of 7 consecutive days.  

 

It was observed that lower dose (250mg/kg) of AERC did 

not produce significant anticonvulsant effect as compared 

to control. But medium and high doses (500&750mg/ kg 

respectively) exhibited a significant anticonvulsant effect 

by increasing onset time of seizures and reducing the 

duration of tonic-clonic seizures (Figure 1 – 3).  

Strychnine induced convulsion: The mice were given 

Ruta chalepensis leaf extract in three different doses 

considerably delayed the onset of seizures. Of the 6 

animals tested, 4 survived at the higher dose (750 mg/kg), 

2 survived at the intermediate level (500 mg/kg), and none 

survived at the lower dose. For higher and moderate doses, 

the percentage protection was 66.66 and 33.33 respectively 

(Table 3). 

Table 3: Effect of AERC on Strychnine induced convulsion.  

All values are expressed as (Mean ± S.E.M), n=6, **p≤0.01, ***p≤0.001 when compared with control. (Statistically 

analyzed by ANOVA allowed by Dunnett’s-test). 

Contro
l

Diazepam

AERC (2
50mg/kg)

AERC (5
00 m

g/kg)

AERC (7
50 m

g/kg)
0

100

200

300

400

500

                  Effect of AERC on Strychnine induced convulsion

Treatment

On
se

t o
f t

on
ic

-c
lo

ni
c 

co
nv

ul
si

on
(s

ec
)

Control

Diazepam

AERC (250mg/kg)

AERC (500 mg/kg)

AERC (750 mg/kg)

 

Fig. 1: Effect of AERC on Strychnine induced convulsion. 

AERC leaves were tested for their ability to prevent 

convulsions induced by strychnine in mice. A chronic 

study was conducted using three different doses of AERC 

(250mg/kg, 500mg/kg, and 750mg/kg), administered once 

daily for seven consecutive days. The lower dose did not 

show a significant anticonvulsant effect compared to the 

control group, but the medium and high doses (600mg/kg 

and 800mg/kg) demonstrated a significant effect by 

Group Treatment Dose No. of animals Parameters to be assessed 

Group I (control) Saline + Pentylenetetrazole 10ml/kg + 80mg/kg 6  

Percentage inhibition of 

seizures relative to 

control 

Group II (standard) Diazepam + Pentylenetetrazole 10mg/kg + 80mg/kg 6 

Group III (Low Dose) AERC + Pentylenetetrazole 250mg/kg + 80mg/kg 6 

Group IV (Moderate Dose) AERC + Pentylenetetrazole 500mg/kg + 80mg/kg 6 

Group V (High Dose) AERC + Pentylenetetrazole 750mg/kg + 80mg/kg 6 

Treatment Onset of tonic clonic convulsion Mean ± 

SEM 

Status of animal (1hr) (No. of animals 

Alive) 

% Protection 

Control 254.50 ± 0.29 0 0 

Diazepam 481.72 ± 0.87*** All 100 

AERC (250mg/kg) 252.83 ± 0.57 0 0 

AERC (500mg/kg) 276.55 ± 0.89 2 33.33 

AERC (750mg/kg) 276.50 ± 1.52** 4 66.66 



Bhat R et al.                              Protective Effects of Ruta Chalepensis Linn. Extracts on drug-induced seizures 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 116  

delaying the onset of seizures and reducing the duration of 

tonic-clonic seizures. The protective effect of the medium 

and high doses of ARC was 33.33% and 66.66% for up to 

one hour after administration, respectively, while the lower 

dose showed no protective effect. The standard drug 

diazepam (5mg/kg) exhibited a significant anticonvulsant 

activity and provided 100% protection (Table 4). 

PTZ induced convulsion: Ruta Chalepensis leaf extract, 

750 mg/kg, considerably delayed the onset of seizures in 

mice treated in the PTZ-induced convulsion model, and it 

significantly decreased the length of tonic-clonic seizures 

for both the moderate and higher dosages of test as well as 

standard. In the test with the higher dose, only two of the 

six animals showed signs of survival, whereas one animal 

did so for the test with the moderate level and none at the 

low amount. The percentage of protection was 33.33 for 

higher doses and 16.66 for moderate doses, respectively.

Contro
l

Dia
ze

pam

AERC (2
50m

g/k
g)

AERC (5
00 m

g/k
g)

AERC (7
50 m

g/k
g)

0

200

400

600

800

1000

Effect of ARC on PTZ induced convulsion

Treatment

O
ns

et
 o

f s
ei

zu
re

 (s
ec

)

Control

Diazepam

AERC (250mg/kg)

AERC (500 mg/kg)

AERC (750 mg/kg)

Fig. 2: Effect of AERC on PTZ induced convulsion.  

Contro
l

Diaze
pam

AERC (2
50m

g/k
g)

AERC (5
00 m

g/k
g)

AERC (7
50 m

g/k
g)

0

10

20

30

40

50

Effect of ARC on PTZ induced convulsion

Treatment

D
ur

at
io

n 
of

 to
ni

c-
cl

on
ic

 s
ei

zu
re

 (s
ec

)

Control

Diazepam

AERC (250mg/kg)

AERC (500 mg/kg)

AERC (750 mg/kg)

Fig. 3: Effect of AERC on PTZ induced convulsion 

Table 4: Effect of AERC on PTZ induced convulsion.  

Treatment Onset of Seizure 

(Sec) Mean± SEM 

Duration of Tonic Clonic 

Seizure (Sec) Mean± SEM 

Status of Animal (No. of 

Animals Alive) 

  % Protection(1hr) 

Control 660.17 ± 4.01 38.83 ± 1.85 0 0 

Diazepam 924.67 ± 2.65*** 6.50 ± 0.76*** All 100 

AERC (250mg/kg) 663.33± 2.30 34.00 ± 1.39 0 0 

AERC (500mg/kg) 671.67 ± 4.02 28.5 ± 0.99*** 1 16.66 

AERC (750mg/kg) 689.83 ± 4.40*** 23.50 ± 1.05*** 2 33.33 

All values are expressed as (Mean ± S.E.M), n=6, ***p≤0.001 when compared with control. (Statistically analyzed by 

ANOVA allowed by Dunnett’s-test). 

AERC leaves were tested for anticonvulsant activity in 

mice using the PTZ induced convulsion model. Three 

different doses of ARC (250mg/kg, 500mg/kg, and 

750mg/kg) were administered once daily for seven 



Bhat R et al.                              Protective Effects of Ruta Chalepensis Linn. Extracts on drug-induced seizures 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 117  

consecutive days. The lower dose did not show significant 

anticonvulsant activity compared to the control group, but 

the medium and high doses showed a significant effect by 

delaying the onset of seizures and reducing the duration of 

tonic-clonic seizures. The protective effect of the medium 

and high doses of AERC was 83.33% and the lower dose 

was 33.33% for up to one hour after administration. The 

standard drug, diazepam (1mg/kg), exhibited significant 

anticonvulsant activity and provided 100% protection. 

Ruta chalepensis is an ancient medicinal plant still used 

in the traditional medicine of many countries as a laxative, 

anti-inflammatory, analgesic, anti-spasmodic, 

abortifacient, anti- epileptic, emmenagogue and for 

dermatopathy treatment. The main aim of this work is to 

investigate the antiepileptic activity of Ruta chalepensis 

aqueous leaf extracts in experimental animals [10]. The 

antiepileptic activity of the Ruta chalepensis leaf extracts 

has not yet been studied for anti-convulsant activity. He 

exact mechanism underlying antiepileptic activity of Ruta 

chalepensis Linn. Leaf extract is not clear but it may be 

apparently related to active compounds present in Ruta 

chalepensis are reviewed. Chemical studies have reported 

the presence of several flavonoids, alkaloids, phenols, 

amino acids, furanocoumarins and saponins in the leaves 

of Ruta chalepensis Linn [11]. The results indicate that 

aqueous leaf extract of Ruta chalepensis Linn. May have 

an antiepileptic like effect. However further experiments 

are necessary to confirm this hypothesis. 

Future Prospectives 

Research in the field of plant-based anticonvulsants has 

yielded promising results in recent years. However, it is 

important to note that this research is still in its early 

stages, and more studies are needed to determine the safety 

and efficacy of these treatments. Nonetheless, the 

discovery of new plant-based anticonvulsants has the 

potential to lead to the development of new and more 

effective therapies for convulsions. Further research and 

clinical trials will be necessary to fully evaluate the 

therapeutic potential of these plant-based treatments. 

CONCLUSION 

In six meticulously planned studies involving laboratory 

animals, the aqueous leaf extract of Ruta chalepensis Linn. 

Showed strong anticonvulsant action, indicating the 

possibility of using it to treat different kinds of epilepsy. 

By using actual measurement and statistical validation, the 

results were validated and observer bias was removed. 

Also, the chemical components of the extract were  

determined. These findings give a scientific basis for the 

extract's therapeutic efficacy even if more research on 

human patients is required. 

Acknowledgements: We are thankful to Research guide, 

Principal and Management of Srinivas college of 

Pharmacy, Mangalore for providing all the necessary 

facilities to carry out this research work. 

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How to cite this article: Ramdas Bhat, Hemalatha C 

H, A R Shabaraya. Exploring the protective effects of 

aqueous extracts of Ruta Chalepensis Linn. On drug 

induced seizures in animal models. Indian J Pharm 

Drug Studies. 2023: 2(3) 113-117. 

Funding: None          Conflict of Interest: None Stated 

 


