IN-VITRO STUDY OF ANTICONVULSANT ACTIVITY ON THE FRUITS EXTRACTS OF COCCINIA INDICA (WIGHT &ARN.) INDUCED MAXIMAL ELECTROSHOCK SEIZURE (MES) MODEL IN RATS

IN-VITRO STUDY OF ANTICONVULSANT ACTIVITY ON THE FRUITS EXTRACTS OF COCCINIA INDICA (WIGHT &ARN.) INDUCED MAXIMAL ELECTROSHOCK SEIZURE (MES) MODEL IN RATS

Mayank Kumar*¹,Shashi Alok ¹, Sanjay Kumar Jain¹and Amita Verma ²

Department of Pharmacognosy, Institute of Pharmacy, Bundelkhand University ¹, Jhansi, Uttar Pradesh, India

Department of Pharmaceutical Sciences, Faculty of Health Sciences, Sam Higginbottom Institute of Agriculture, Technology and Sciences-Deemed University ², Allahabad, Uttar Pradesh, India

ABSTRACT:

Aim of the study:In-vitro study of Anticonvulsant activity on the fruits extracts of Coccinia indica (Wight &Arn.) induced maximal electroshock seizure (MES) model in rats.

Materials & methods:  For the assessment of anticonvulsant activity, the animals were divided into five groups of five animals each albino rats. Group I received Normal saline, Group II received Phenytoin, Group III received 200 mg/kg of CIEE suspended in Tween 80, Group IV received 400 mg/kg of CIEE suspended in Tween 80, and Group V received 600 mg/kg of CIEE suspended in Tween 80. The results are expressed in mean ± S.E.M. (n=5) Statistical analysis was done by one way ANOVA, followed Dunnett multiple comparison test vs. control. P<0.01 was considered as statistically significant.

Results: In the present In vitro study, an anticonvulsant effect of ethanolic extracts of fruits Coccinia indica Wight & Arn. has been determined duration of HLTE 14.96, 3.49, 9.68, 8.20, 4.90 sec. The CIEE exhibited maximum anticonvulsant activity at 4.90 sec. at 600mg/kg (P<0.01) and it was significant when compared with control. In the present study, CIEE at the dose of 600mg/kg shows potent anticonvulsant activity as compared to 200mg/kg & 400mg/kg. The ethanolic extract of CI possesses potent anticonvulsant effect against MES induced and it is evidenced by its good anticonvulsant activity of 600 mg/kg against control and as positive control groups.

Conclusions:  In conclusion these observations suggest that 600 mg/kg doses of HLTE exhibit potent anticonvulsant activity.

Coccinia indica, Anticonvulsant, maximal electroshock seizure, Diazepam

INTRODUCTION:The use of plants and plant products as medicines could be traced as far back as the beginning of human civilization. Medicinal plants are a source of great economic value all over the world.

Nature has bestowed on us a very rich botanical wealth and a large number of diverse types of plants grow in different parts of the country ¹. Herbal medicine is still the mainstay of about 75-80% of the whole population and the major part of traditional therapy involves the use of plant extract and their active constituents. Following the advent of modern medicine, herbal medicine suffered a setback, but during last two or three decades, advances in photochemistry and in identification of plant compounds, effective against certain diseases have renewed the interest in herbal medicines ².

Nowadays multiple drug resistance has developed due to the indiscriminate use of commercial antimicrobial drugs commonly used in the treatment of infectious disease. In addition to this problem, antibiotics are sometimes associated with adverse effects on the host including hypersensitivity, immune-suppression and allergic reactions ³. This situation forced to search for new antimicrobial substances. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases from medicinal plants.

Antimicrobials of plant origin have enormous therapeutic potential ⁴. They are effective in the treatment of infectious diseases while simultaneously mitigating many of the side effects that are often associated with synthetic antimicrobials ⁵. The beneficial medicinal effects of plant materials typically result from the combinations of secondary products present in the plant. In plants, these compounds are mostly secondary metabolites such as alkaloids, steroids, tannins and phenol compounds, flavonoids, steroids, resins fatty acids gums which are capable of producing definite physiological action on body ⁶.

Coccinia indica belongs to the family Cucurbitaceae. It is growing wild throughout India and also cultivated in various parts of India. It is commonly known as kundru ^[7]. The whole plant is traditionally used for various medicinal purposes. Leaves of this plant are used in Indian folk medicine for treatment of number of ailments including diabetes, wounds, ulcers, inflammation, in eruptions of skin, fever, asthma and cough. Earlier scientific investigation of Coccinia indica showed that the crude extract has hepatoprotective ^{8, 9 10, 11, 12, 13}, anti-diabetic hypolipidemic ^{14, 15, 16, 17}, anti-bacterial ¹⁸ and anthelmintic activity ^19, analgesic and antipyretic activity ^20, Wound healing activity ^21, anti-inflammatory ^{22, 23} though the plant has been reported for many biological activities, no scientific data available to identify the genuine sample.

The present investigation was therefore taken up to establish identity of fresh and dried fruits anticonvulsant activity for the standardization of the drug.

MATERIALS AND METHODS:

Collection and Authentication: The fresh fruits of wildly growing plant Coccinia indica were collected from the field areas of eastern Uttar Pradesh region during the month of February, 2009. For identification and taxonomic authentication, sample of plant material was given to National Vrakshayurveda Research Institute (NVRI) Jhansi, India. The text report from National Vrakshayurveda research institute, Jhansi, India and confirmed the authenticity of plant material sample was Coccinia indica with voucher Specimen no. NVRI -SOP-20932, 01-09-2012. Collected plants were shade-dried and coarsely powdered. Fruits ofCoccinia indica Wight & Arn. and authentication specimen record card given respectively in figure 1 & 2.

FIGURE 1: FRUITS OF COCCINIA INDICA WIGHT & ARN.

FIGURE 2: AUTHENTICATION OF SPECIMEN RECORD CARD

Extraction of Plant Materials: 250 g coarse powder of air dried fruits of Coccinia indica were packed in muslin cloth and subjected to soxhlet extractor for continuous hot extraction with petroleum ether and ethanol for 8 hrs separately. Then the each extracts were filtered and filtrate was evaporated to dryness.

Preliminary phytochemical studies: Preliminary qualitative chemical investigations of ethanolic extract of Coccinia indica fruits were performed to know the presence of Carbohydrates, Glycosides, Alkaloids, Tannins, Flavonoids, triterpenoids. Further thin layer chromatography was performed to confirm their presence.

Chemical (Drugs & solutions) and apparatus: The following drugs and chemicals were used. Drugs: Phenytoin (Sain medicaments Pvt. Ltd., Hyderabad) and Chemicals: Tween 80 (Chemical laboratory Pharmacy, BU University Jhansi), Normal Saline (0.9% NaCl solution). The plant extract was suspended in Tween 80 and subjected for anticonvulsant. Phenytoin was also dissolved in Tween 80.

Preparation of Dose: The ethanolic extract of Coccinia indica was freshly dissolved in a suitable amount of Tween 80 and was administered at the doses of 200, 400 and 600 mg/kg body wt. p.o. in rats. One hour after p.o. administration, the animals were submitted to the activity. Phenytoin (25mg/kg b. wt.) were dissolved in Tween 80 immediately prior to use and given i.p. Normal saline administered in a volume of 10 ml/kg b.wt.                                                                                                                                                                                                                                                                                                                                                        he purpose of control and supervision of experiments on animals (CPCSEA) with registration number 716/02/a/CPCSEA.

Acute toxicity studies:

1. Acute oral toxicity: Acute toxicity study was performed for the extract according to the acute toxic classic method as per OECD guidelines (OECD Guidelines for the testing of chemicals, 2001). Swiss albino rats of either sex were used for acute toxicity study. The method of Up and Down was used to determine the dose. The animals were kept fasting for overnight providing only water, after which the extracts were, administered orally 100, 300, 500, 2000, 5000 mg/kg p.o. dose and percent mortality was observed 24 hrs then 72 hrs and thereafter once daily for 14 days.

The ethanolic extract of fruits of Coccinia indica Wight & Arn.  did not cause any mortality up to 5000 mg/kg during the observation period of 24 hrs then 72 hrs and thereafter once daily for 14 days and were considered as safe. The oral LD₅₀ of the ethanolic extract estimated in mice must be > 5000 mg/kg p.o.

Animal Groups: For the assessment of anticonvulsant activity, the animals were divided into five groups of five animals each albino rats.

Group I received Normal saline.

Group II received Phenytoin.

Group III received 200 mg/kg of CIEE suspended in Tween 80.

Group IV received 400 mg/kg of CIEE suspended in Tween 80.

Group V received 600 mg/kg of CIEE suspended in Tween 80.                             

Assessment of anticonvulsant activity: Corneal electrodes were used for bilateral delivery of electrical stimulus. Electroconvulsive shock (50 mA for 0.2 sec) was delivered through corneal electrode to induce Hind Limb Tonic Extensor (HLTE) phase in rats. There are five phases observed in mice after giving maximal electroshock.

The five phases are;

(i)     Flexor

(ii)   Extensor

(iii)Convulsion

(iv)Stupor and

(v)   Recovery or Death are noted and also the time spent by mice in each phase.

Prior to delivery, the current output was checked by using multimeter. The electrical stimulus was applied using a stimulator apparatus (Biocraft Scientific System Pvt. Ltd., Agra, India) for five groups of five animals each. The orientation for the anticonvulsant affect was abolition of HLTE within 10 sec after delivery of the electroshock (Figure 3). Delivery of MES using electroconvulsiometer to mice with the help of corneal electrode (Figure 4) Phases occur after giving electroshock (Figure 5) Graphical representation of anticonvulsant activity of Coccinia indica on maximal electroshock (MES) induced seizures in rats.

FIGURE 3: DELIVERY OF MES USING ELECTROCONVULSIOMETER TO MICE WITH THE HELP OF CORNEAL ELECTROD

FIGURE 4: PHASES OCCUR AFTER GIVING ELECTROSHOCK

FIGURE 5: GRAPHICAL REPRESENTATION OF ANTICONVULSANT ACTIVITY OF COCCINIA INDICA ON MAXIMAL ELECTROSHOCK (MES) INDUCED SEIZURES IN RATS

 Assessments for anticonvulsant activity Control, Standard drug (Phenytoin), Coccinia indica ethanolic extract by MES model are given in table 1, 2, 3 respectively.Effect of Coccinia indica extract on MES induced seizures in rats are also given in table 4 respectively.

TABLE 1: ASSESSMENT FOR ANTICONVULSANT ACTIVITY OF CONTROL BY MES MODEL

R = Recovery, D = Death, A = Absent

TABLE 2: ASSESSMENT FOR ANTICONVULSANT ACTIVITY OF STANDARD DRUG (PHENYTOIN) BY MES MODEL

R = Recovery, D = Death, A = Absent

TABLE 3: ASSESSMENT FOR ANTICONVULSANT ACTIVITY OF COCCINIA INDICA ETHANOLIC EXTRACT BY MES MODEL

R = Recovery, D = Death, A = Absent

TABLE 4: EFFECT OF COCCINIA INDICA EXTRACT ON MES INDUCED SEIZURES IN RATS

 Statistical analysis: The results are expressed in mean ± S.E.M. (n=5) Statistical analysis was done by one way ANOVA, followed by Dunnett multiple comparison test vs. control.  P<0.01 was considered as statistically significant.

RESULTS: In the present in vitro study, an anticonvulsant effect of ethanolic extracts of fruits Coccinia indica Wight & Arn. has been determined Duration of HLTE 14.96, 3.49, 9.68, 8.20, 4.90 sec. The CIEE exhibited maximum anticonvulsant activity at 4.90 sec. at 600mg/kg (P<0.01) and it was significant when compared with control. In the present study, CIEE at the dose of 600mg/kg shows potent anticonvulsant activity as compared to 200mg/kg & 400mg/kg.

Anticonvulsant can act on corneal electrodes were used for bilateral delivery of electrical stimulus. Electroconvulsive shock (50 mA for 0.2 sec) was delivered through corneal electrode to induce Hind Limb Tonic Extensor (HLTE) phase in rats. The ethanolic extract of CI possesses potent anticonvulsant effect against MES induced and it is evidenced by its good analgesic activity of 600mg/kg against control and as positive control group.

DISCUSSION: The present work was to carry out with the objective of ethanolic extract of fruits Coccinia indica Wight & Arn., Phytochemical investigation, evaluation of the anticonvulsant activity (in-vitro) against induced maximal electroshock seizure (MES) model in rats. The ethanolic extract of CI possesses potent anticonvulsant effect against MES induced and it is evidenced by its good analgesic activity of 600mg/kg against control and as positive control group.

The preliminary phytochemical screening of fruits Coccinia indica Wight & Arn. shows the presence of steroids and flavanoids compounds as major active principle constituents in ethanolic extract of fruits Coccinia indica Wight & Arn.. Many of flavonoids and steroids compounds have been reported for hepatoprotective activity.Therefore, there is possibility that ethanolic extract of Coccinia indica Wight & Arn. fruits may possess the anticonvulsant activity.

CONCLUSION: It  can  be  concluded  that  the  present  study  on  C. indica  fruits  can  serve  as  an  important  source  of  information  to  the reduction in oxalate excre­tion was observed on CIEE treatment. This decreased excretion of oxalate may be due to the inhibition of formation of oxalate by the plant extract. The ethanolic extract of CI possesses potent anticonvulsant effect against MES induced and it is evidenced by its good analgesic activity of 600mg/kg against control and as positive control group.

Conflict of interest statement: We declare that we have no conflict of interest.

REFERENCES:

1. Ahmed L, Mohammed Z, Mohammed F. Screening of some Indian medicinal plants for their antimicrobial properties.  J Ethnopharmacol 1998; 62: 183-193.
2. Arora D, Kaur J. Antimicrobial activity of spices. International J Antimicrobial Agents 1999; 12: 257-262.
3. Gupta C, Amar P, Ramesh G, Uniyal C, Kumari A.  Antimicrobial activity of some herbal oils against common food-borne pathogens.  African J Microbiol Res 2008; 2: 258-261.
4. Werner F, Okemo P, Ansorg R. Antibacterial activity of East African Medicinal plants. J Ethnopharmacol 1999; 60: 79-84.
5. Perumalsamy R, Ignacimuthu S. Antibacterial activity of some folklore medicinal plants used by tribals in Western Ghats of India.  J Ethnopharmacol 2000; 69: 63-71.
6. Bishnu J, Sunil L, Anuja S. Antibacterial Property of Different Medicinal Plants Ocimum sanctum, Cinnamomum zeylanicum, Xanthoxylum armatum and Origanum majorana Kathmandu University. J Sci Engineering and Technol 2009; 5: 143-150.
7. Wasantwisut E, TharaV. Ivy Gourd  (Coccinia grandis Voigt, Coccinia cordifolia, Coccinia indica) in Human Nutrition and Traditional Applications, Plants in Human Health and Nutrition  Policy. World Reviews of Nutrition and Dietics, Basel, Karger 2003; 91: 60-66.
8. Vadivu R, Krithika A, Biplab C, Dedeepya SP, Lakshmi KS. Evaluation of Hepatoprotective Activity of the Fruits of Coccinia grandis Linn. International J Health Res 2008; 3: 63-168.
9. Rao GM Mohana, Vijayakumar M, RaoCh V, Rawat AKS, Mehrotra S. Hepatoprotective effect of Coccinia indica against CCl₄ induced Hepatotoxicity. Natural Product Sciences 2003; 1: 13-17.
10. Shyam BK, Gnanasekaran D, Jaishree V, Channabasavaraj KP. Hepatoprotective activity of Coccinia indica leaves extract. Int J Pharm Biomed Res 2010; 4: 154-156.
11. Ushasri B, Kiranmai M, Ibrahim M. Evaluation of Coccinia indica Mucilage as suspending agent in paracetamol. International journal of Drug Formulation and Research 2011; 6: 237-247.
12. Maheswari C, Babu P, Meenakshi. Protective effect of Coccinia indica leaf extract against alcohol combined with carbon tetra chloride and paracetamol induced liver damage in rats. International Journal of Pharmaceutical Sciences and Research 2011; 10: 2660-2664.
13. Shivaji PG, Chandrasekhar Rao MV. Antihepatotoxic Activities of Ci Compound ß Sitosterol Isolated from Fruits and Leaves of Coccinia indica. Indian Journal of Pharmaceutical Education and Research 2012; 1: 4- 8.
14. Shakya VK. Antidiabetic activity of Coccinia indica in streptozotocin induced diabetic rats. Asian Journal of Chemistry 2008; 8: 6479-6482.
15. Mallick C, Mandal S, Barik BB, Atanu, Ghosh D. Protection of testicular dysfunctions by MTEC. A formulated herbal drug, in streptozotocin induced diabetic rat. Biological & Pharmaceutical Bulletin 2007; 1: 84-90.
16. Manish G,Goutam KJ, Jha AK, Umashanker M. Pharmacognostic antihyperglycemic study of Coccinia indica. International Journal of Phytomedicine 2010; 2: 36-40.
17. Versha P, Akanksha S, Ishan D,Deepak K. Combined effect of Coccinia indica (Wight &Arn) and Salvadora oleoides (Decne) on blood glucose level and other risk factors associated with Type-2 diabetes mellitus in alloxan induced diabetic. International Journal of Pharmacy and Pharmaceutical Sciences 2012; 4:79-84.
18. Arshad H, Shadma W, Iffat Z, Sarfaraj HMD. Antibacterial Activity of the Leaves of Coccinia indica (W. and A) of India. Advances in Biological Research 2010; 5: 241-248.
19. Yogesh S, Prashant S, Priya S, Sonal D, Sourabh SB. Evaluation of Anthelmintic Activity of Coccinia indica (fruits). J Chem Pharm Res 2011; 1: 488- 491.
20. Niazi Junaid, Singh Parabhdeep, Bansal,Yogita, Goel RK. Anti-inflammatory, analgesic and antipyretic activity of aqueous extract of fresh leaves of Coccinia indica. Inflammopharmacology 2009; 4: 239-44.
21. Bambal VC, Wyawahare NS, TuraskaAO, Deshmukh TA. Evaluation of Wound healing activity of herbal gel containing the fruit extracts of Coccinia indica Wight & Arn. (Cucurbitaceae). International Journal of Pharmacy and Pharmaceutical Sciences 2011; 4: 319-324.
22. Sumana C, Aritra C. Proximate analysis, phyto-chemical screening and anti-inflammatory activity of Coccinia indica. International Journal of Pharmaceutical, Chemical and Biological Science. 2012; 3: 299-304.
23. Vaishali CB, Nitin SW, Sandip PC, Tushar AD, & Vijay RP. Phytochemical Investigation and Anti-Inflammatory Activity of Coccinia indica Wight and Arn. (Cucurbitaceae) Fruits. Lat Am J.Pharm 2010;5: 820- 4.
    

    ---

    How to cite this article:

    Kumar M, Shashi Alok, Jain SK and Verma A: In-vitro study of anticonvulsant activity on the fruits extracts of Coccinia indica (Wight &Arn.) induced maximal electroshock seizure (MES) model in rats. Int J Pharm Sci Res 2013; 4(11): 4319-24. doi: 10.13040/IJPSR. 0975-8232.4(11).4319-24

    ---

    All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.

    ---