INTRODUCTION:

These are a group of CNS disorders characterized by paroxysmal cerebral dysrhythmia, manifesting as brief episodes (seizures) of loss or disturbance of consciousness, with or without characteristic body movements (convulsions), sensory or psychiatric phenomena.

Epilepsy is a chronic disorder of the brain that affects people worldwide. As per WHO, epilepsy is characterized by recurrent seizures, which are brief episodes of involuntary movement that may involve a part of the body (partial) or the entire body (generalized), and are sometimes accompanied by loss of consciousness and control of bowel or bladder function¹. The strange behaviour caused by some seizures has contributed through the ages to many superstitions and prejudices. From greek word attack, the word epilepsy is derived.

Traditional systems of medicines are popular in developing countries and upto 80% of the population relies on traditional medicines/folk remedies for their primary health care need². Hence, there is a need to discover an alternative agent from natural sources³. Juglans regia used as a herbal medicine and is well known for its traditional uses such as antioxidants, neuropeotective, anticancer, cardioprotective etc. Various studies shows that the active principle diterpene alkaloids having a crucial role in treatment of epilepsy^4,5. Juglans regia is rich in alkaloids, phenols.

Fig No.1 Comparison of normal and seizure activity of brain

Epilepsy accounts for 0.6%, of the global burden of disease, a time-based measure that combines years of life lost due to premature mortality and time lived in less than full health. In terms of health care needs, premature death and lost work productivity, epilepsy has significant economic implications. The search for agents with anticonvulsant activity with more selectivity and lower toxicity continues to be an area of investigation in future².

Having a seizure does not necessarily mean that a person has epilepsy. Only when a person has had two or more seizures is he or she considered to have epilepsy³.

MATERIAL AND METHODS:

Material:

Plant collection and authentication:

The fruit of Juglans regia was collected from the local market of Pusad. The fruit was identified by Dr R.J. Mandade.

Authentication of plant was done by Dr Sakhare sir Ayurved Mahavidyalay Pusad.

Extraction of plant (Juglans regia fruit):

Juglans regia fruit were cracked then separated into two parts (septum and kernel), then coarsely powdered and passed through mesh sieves. The powdered material (kernel)was extracted with ethanol using soxhlet apparatus at room temperature for 24hrs. The solvent was evaporated by rotary evaporation at 35degree Celsius, yielding a dark red colour mass and stored at 4 degree Celsius.

Drugs:

Pentylenetetrazole-60mg/kg, Diazepam- 1mg/kg, Juglans regia-100mg/kg

Experimental animal:

Mice- (Swiss albino mice) of both sex weighing between 30-40gm.

Fig No.2 Mice

PTZ- induced convulsion in mice:

1. Divide the animal into three groups (PTZ, STD, Extract) consisting of 3mice.

2. Weigh and number them.

3. Inject pentylenetetrazole to first group animal.

4. Note the onset of action of the jerk movement of the body, duration of jerk.

5. Inject diazepam to the standard group, injetct extract to the test group.

6. After 30min inject PTZ to these animal which received STD and extract.

7. Note the onset and the severity of the convulsions.

8. Note eighter delay or complete abolition of convulsion in the mice treated with STD, extract.

MES- induced convulsion in mice:

1. Divide the animal into three groups (control, STD, extract) each consisting of 3 mice.

2. Weigh and number them.

3. Hold the animal (control) properly place electrodes on ear pinna.

4. Apply prescribed current (12mA for 0.2 sec)

5. Note the time sec spent by the animal in each phase of the convulsion a) tonic flexion, b) tonic extensor phase, c) clonic convulsion, d) stupor, e) recovery or death.

6. Inject diazepam intraperitonially to the STD group. Extract to the test group.

After 30 min subject the animal to electro convulse meter and hold the animal properly place the electrodes on the ear pinna. Apply prescribed current

(12mA for 0.2 sec)

7. Note down the reduction in time or abolism of tonic extensor and flexion phase of MES.

Grip strength test-Rota rod:

1. Dividethe animal into three groups (control, STD, extract) each consisting of 3 mice.

2. Weigh and number them.

3. Turn on Rota rod. Select speed (25rpm).

4. Place animal one by one on rotating rod.

5. Inject diazepam intraperitonially to the STD group. Extract to the test group.

6. Note down the ‘fall of time’

7. Compare the fall of time of animals in control, STD, extract treated groups.

RESULTS:

Preliminary phytochemical analysis of plant Juglans regia

Table No. 1 Preliminary phytochemical analysis of plant Juglans regia

Sr No	Constituents	Presence/Absence
1	Alkaloids	+
2	Carbohydrate	+
3	Glycoside	+
4	Phenolic compound	+
5	Flavonoids	+
6	Acidic compound	-

PTZ induced seizure:

Table No.2: Effect of ethanolic fruit extract of juglans regia on PTZ Induced Seizure in mice

Groups	Treatment	Onset of Seizure (Min)	Recovery / Death
Group I	PTZ Treated 60 mg/kg	3.0±0.57	Death
Group II	STD 1mg/kg	23.67±0.88^***	Recover
Group III	Extract 100mg/kg	28.0±1.0^***	Recover

Value expressed in mean ± SEM [n=3]; statistical analysis done by one-way ANOVA which is followed by Tuckey test

*indicate the comparison of inducer group with treatment group (*** p<0.0001)

Fig No.3 Effect of ethanolic fruit extract of juglans regia on PTZ Induced Seizure in mice

· MES induced seizure:

Table No.3 Effect of ethanolic fruit extract of juglans regia in MES Induced Seizure in mice

Treatment	Duration of Flexion	Duration of Extension	Duration of stupor	Recovery / Death
Control	8.33±0.88	17.33±0.66	28±1	Recover
Standard 1mg/kg	2.33±0.33^**	8.66±0.66^***	Nil	Recover
Extract Treated 100mg/kg	5±0.57^*	12±1.15^*	Nil	Recover

Value expressed in mean ± SEM [n=3]; statistical analysis done by one-way ANOVA which is followed by Tuckey test

*indicate comparison of Treatment group with control group (* p<0.05, ** p<0.001, ***p<0.0001)

Fig No. 4: Effect of ethanolic fruit extract of juglans regia in MES Induced seizure in mice

Fig No. 5 Effect of ethanolic fruit extract of juglans regia in

MES induced seizure in mice

Fig No. 6 Evaluation of grip strength in different treated groups

DISCUSSION:

Juglans regia used as a herbal medicine and is well known for its traditional uses such as antioxidant, neuroprotective etc^7,8. Various studies shows that the active principle of phenolic compound having a crucial role in treatment of epilepsy. Since Juglans regia have not been studied for its antiepileptic activity, the present study was aimed to evaluate the antiepileptic activity ethanolic extract of Juglans regia fruit.

Epilepsy may develop because of an imbalance of nerve signaling chemicals called neurotransmitters. In case of epilepsy, there may be abnormally high level of excitatory neurotransmitters (glutamate) that increase neuronal activity, while abnormally low level of inhibitory neurotransmitters (GABA) that increase neuronal activity in the brain⁹.

In case of PTZ induced convulsion, the result of the present study shows that the ethanolic extract of Juglans regia, at dose 100 mg/kg significantly reduced the duration and also delayed the onset of convulsion and P value was found to be P<0.0001.

The maximal electroshock induced convulsion in animals represents grand mal type of epilepsy. The tonic extensor phase and flexion phase is selectively abolished by the drugs effective in generalized tonic clonic seizure¹⁰.

The maximal electroshock induced convulsion, the result of the present study shows that the ethanolic extract of Juglans regia at dose 100 mg/kg significantly delayed and reduced the onset of flexion and extension, P value was found to be *p<0.05, **p<0.001, ***p<0.0001.

In case of Grip strength test – the result of the present study shows that the ethanolic extract of Juglans regia, at dose 100mg/kg significantly reduces the fall off time and P value Was found to be **p<0.001, ***p<0.0001.

Result shows that the extract gives the protection against both physical and chemical method that is in PTZ induced seizure and MES induced seizure.

CONCLUSION:

Epilepsy is a neurological disorder that affects a wide range of people throughout the world. Approximately 30% of the patients continue to have seizures with current antiepileptic drug therapy. Natural products from folk remedies have contributed significantly in the discovery of modern drugs and can be an alternative source for the discovery of antiepileptic drugs with novel structures and better safety and efficacy profiles.

The ethanolic extract of Juglans regia delayed the onset and reduced the duration of convulsion in MES and PTZ induced convulsion models and can be used as an adjuvant therapy against cognitive deficit in convulsions.

From the result obtain after successful research work following conclusion have been made,

The results of the ethanolic extract of Juglans regia reveals it possess anticonvulsant activity either by blocking sodium channel or by enhancing GABA receptor mediated inhibitory transmission.

To conclude, the result of research study, seizures was induced by PTZ in animal model, the extraction shows the highly significant anti-convulsant activity.

The seizures induced by MES in animal model the extract gives the protection against the shocks and show moderately significant action.

CONFLICT OF INTEREST:

There is no conflict of interest between the authors or anybody with respect to research presented in article.

ACKNOWLEDGEMENTS:

The authors would like to thank Principal of SNIOP and Dr R.J. Mandade, Department of Pharmacology, Prof. Pranay Uplenchwar for their support and guidance.

REFERENCE:

1. Jain, P., et al., Epilepsy: A neurological cramp. 2013. 5(1): p. 1-5.

2. Tufon, E.N., et al., Evaluating the Management of Epilepsy in the Ngie Community of the North West Region of Cameroon. 2013. 10(15): p. 11.5.

3. Ratna, B., et al., Investigation on the effect of the Drug Levetiracetam combined with Clobazam on MES Model of Epilepsy. 2020. 13(6): p. 2792-2796.

4. Arun, J., et al., Phytochemical and Pharmacological screening Daucus carota seed for its Anticonvulsant activity by using Pentylenetetrazole Induce Convulsion. 2022. 15(5): p. 2293-2296.

5. Debnath, J., et al., An Experimental Evaluation of Anticonvulsant Activity of Ethanolic Extract of Seeds of Holarrhena antidysenterica In Mice. 2011. 3(1): p. 31-33.

6. Tiwari, R.N., et al., Evaluation of Anticonvulsant Activity of Novel Substituted 2-Mercaptobenzimidazole Derivatives. 2010. 3(2): p. 466-467.

7. Senghani, M.K.J.R. Anticonvulsant Activity of Boswellic acids in Pentylenetetrazole-induced Convulsive mice and Thiopental Sodium induced Sleeping time in Mice. J.O.P. and Technology. 2013. 6(9): p. 1037-1041.

9. Kannadasan, M., et al., Anti-Epileptic Activity of the Hydro-Alcoholic Extract of Erythrina fusca Lour. Bark through Behavioral Studies against the Animal Models of Epilepsy. 2011. 4(2): p. 315-319.

10. Komali, E., et al., Anticonvulsant effect of Bacopa monnieri extracts on Catecholamine metabolism during PTZ-induced Epilepsy in different brain regions of Albino Rat. 2018. 11(4): p. 1592-1598.

11. Gautam, S.K., et al., Synthesis and Anticonvulsant activity of new 5-((1-(2-(4-substituted phenacyl bromide)-2-oxoethyl)-2-phenyl-1H-indol-3yl) methylene) pyrimidine-2, 4, 6 (1H, 3H, 5H)-trione Derivatives. 2021. 14(5): p. 2681-2685.

Received on 10.05.2023 Modified on 05.06.2023

Res. J. Pharmacology and Pharmacodynamics.2023;15(4):155-158.

DOI: 10.52711/2321-5836.2023.00028

Groups	Fall of Time (sec)
Groups	30	60	90	120	150	180
Control	46.67±1.4	52.67±2.6	47±1.4	47.67±1.4	46±3.0	45±1.7
Standard 1mg/kg	15±1.14	15.67±1.4	14.67±0.8	12.33±1.2^***	12±1.1	13.33±1.2
Extract Treated 100mg/kg	20.67±0.8	21±1.1	24±1.7	27.67±1.4^**	27.63±1.2	27.30±1.2