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M-16: Protective Effect of Juice, Ether and Ethyl Acetate Extracts of Benincasa Hispida Fruit in PTZ Induced Seizures in Zebra Fish

Poster Presenter

Deena D'Souza

B. Pharm Student
Bombay College Of Pharmacy
India

Objectives

To explore the effects of Benincasa hispida fruit juice (swarasa), its petroleum ether and ethyl acetate extracts on pentylenetetrazole (PTZ) induced convulsions in zebra fishes (Danio rerio).

Method

Fishes were exposed to a 100mL of juice/extract solutions for 30 min and then to a 100mL PTZ (0.06mM) for 1 min. Each fish was transferred to tank and video recorded for 10 min using Panlab smart 3.0 software and behavior was studied. Visual examination was done to note seizure latencies and type.

Results

Fast swimming was observed in the first 20 seconds after transfer into observation tank and this was attributed to transfer stress. Fast swimming and confusion thereafter were categorized as pre seizure movements and seizures were categorized into 3 stages, stage 1: rapid movement across the tank bottom, stage 2: Slight loss of posture, stillness and collisions with the tank wall and stage 3: cork screw movements, complete loss of posture, turning over. Fishes exposed to PTZ (0.06mM) showed either a high mean max speed or low mean max speed as some fishes showed stage 1 seizures to stage 3 seizures while other fishes showed loss of posture and stillness thus showing an increased (398.45s) or decreased (0.9s) resting time in zone, respectively. Seizures latencies were between 30 seconds-2 minutes. All three stages of seizures were seen. 42.85% of seizures observed were of stage 3 type, 20.4% of stage 2 and 36.75% of stage 1 type. Fishes exposed to the standard carbamazepine solution (10µM) only showed pre seizure movements. Fishes exposed to swarasa (2% v/v) showed only stage 1 seizures and latencies were observed to be between 4:30-6 minutes. Mean speed in zone (7.67 cm/sec) was seen to be only slightly higher than that of fishes exposed to blank solution (6.78 cm/sec). Fishes exposed to ether extract (0.0025% w/v) showed latencies between 3-5 minutes and four out of five fishes showed only stage 1 seizures. Brief stillness (stage 2 seizure) was shown by one fish. Mean average speed (6.39 cm/sec) was seen to be lower than that of blank (6.78 cm/sec) which was due to the occurrence of stage 2 seizure. Of the total seizures, stage 1 seizures accounted for 93.1% and stage 2 for 6.89%. Fishes exposed to ethyl acetate extract (0.0025% w/v) showed seizure latencies between 4 and 5:30 minutes. All except one fish showed only stage 1 seizures. One fish showed stage 2 seizure (loss of posture). 95% of the seizures observed were of stage 1 type and 5% were of stage 2 type.

Conclusion

The results point towards the efficacy of Benincasa hispida fruit in protecting against epileptic convulsions in zebra fishes. The seizure latencies were seen to be delayed, and the intensities were seen to have reduced. Ether soluble and acetate soluble components similarly showed protective action but to a lesser extent than whole fruit juice (swarasa). This could point towards synergism of various components in the fruit playing a role in the exerted activity. Benincasa hispida is a common part of the diet in south Asia and many other cultures and has been used since ancient times for treating various illnesses and ancient Indian Ayurveda texts reveal the same. This study gave us preliminary evidences on the promise of the fruit of Benincasa hispida in treating epilepsy. Thorough preclinical studies are required to consolidate this claim. The coauthors of this project are: Mr. Samyak Bafana, Mr. Nachiket Dandekar, Dr. Anuradha Majumdar(mentor)