Evaluation for CNS Activities of Hexane Extract of Citrullus lanatus Seeds

 

Habibur Rahman1,2*, M. Chinna Eswaraiah2, Anoosha T.2, Nagaveni K.2, Manjula K.2

1Research Scholar, Assam Down Town University, Guwahati, Assam 

2Department of Pharmacology, Anurag Pharmacy College, Kodad, Andhra Pradesh-508206, India

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

 

ABSTRACT:

The present study aimed to evaluate the possible CNS activities of hexane extracts of Citrullus lanatus Seeds. The effect of hexane extracts of Citrullus lanatus Seeds was tested for Anti-depressant activity using Forced Swim Test (FST) and Tail Suspension Test (TST) in mice, Anxiolytic activity using Elevated Plus Maze(EPM) and Dark-light model in mice and effect on motor co-ordination and skeletal muscle using Rota-Rod , Grip Strength test in mice. The hexane extract exhibited significant anti-depressant and anxiolytic activities but the extract does not produces skeletal muscle relaxant activity beside it slightly improved the skeletal muscle activity in animal models. It can conclude that hexane extract of Citrullus lanatus Seeds possess anti-depressant and anxiolytic activity but does not affect on skeletal muscle activity. Finally, it can say that this can be used in CNS disorders like depression and anxiety and does not affect of skeletal muscle activity like synthetic drugs.

 

KEYWORDS -: Citrullus lanatus Seeds, hexane extract, Forced Swim Test, anxiety, skeletal muscle activity


 

INTRODUCTION:

Citrullus lanatus of family Cucurbitaceae is commonly known as water melon and in local name Tarmuz (Hindi), Puchakaya (Telugu). The ripe fruits are edible and largely used for making confectionary. Its nutritive values are also useful to the human health. Fruit is used in cooling, strengthening, aphrodisiac, astringent to the bowels, indigestible, expectorant, diuretic, and stomachic, purifies the blood, allays thirst, cures biliousness, good for sore eyes, scabies and itches and as brain tonic to the brain 1. It also reported having analgesic and anti-inflammatory of seeds2, anti-ulcerative activity3,4, antimicrobial activity5, laxative activity of fruit6, antioxidant of fruit7, hepatoprotective8 and anti-hyperlipidemic9.

 

Literature survey reveals the uses Citrullus lanatus fruit Seeds contain unsaturated fats almost 90%, vitamins, antioxidants, minerals, proteins and phytochemicals like terpinoids, Flavanoids and phenolic compounds10.

 

Oxidative damage to cellular biomolecules such as lipids, proteins and DNA is thought to play a crucial role in the incidence of several chronic diseases including CNS degenerative, anxiety, Alzheimer and cardiovascular etc11-13.

 

Flavnoids and phenolic containing herbal drugs has got increased demand in recent years owing to their potent antioxidant and free-radical scavenging activities14-17.

 

Though Watermelon Fruit  is an excellent source of vitamins, arginine, Carotenoids, lycopene, carbohydrate, sodium, magnesium, potassium and water and consumed widely throughout the world18-19. But the seeds are not in generally given importance though it contain important phytochemicals and hence the present investigation was conducted to study CNS activities of hexane extracts of Citrullus lanatus seeds in experimental animal models.

 

MATERIAL AND METHODS:

Plant material and extraction procedures:

Citrullus lanatus fruit were purchased local market from Kodad, Andhra Pradesh  and seeds were collected from fruits and  was authenticated by Prof. Dr. K. Madhava Chetty,Taxonomist, SVU University, Chithoor, Andhra Pradesh (India).The air dried seeds were made into coarse powder and extracted with n-Hexane using Soxlet Apparatus  and percentage yield were calculated.

 

Preliminary Phytochemical Analysis

The  hexane extract of  Citrullus lanatus were tested for different phytoconsituents like alkaloids, glycosides, saponinins, tannins, terpinoids, phenolic compounds, protein, carbohydrates using standard procedures20.

 

Experimental Animals

Mice of either sex weighing 150-200 g of body weight were used in experiment. Animals were obtained from Anurag Pharmacy College, Kodad. Animals were kept under standard conditions at 23-250C for 12 hr light/dark cycle and given standard pellet diet and water. The animals were accustomed to the laboratory conditions for a week prior to the experimentation. The fresh diet and water for the animals has to be supplied daily to the animals. The condition of the animals has to be supervised daily till the completion of the experiment.

 

Vehicles and Preparation of Doses

To prepare the dosage forms extract of Citrullus lanatus seed is made a suspension with 1% tween 80. The dose in required concentration was administered at 1ml/100g body weight of the animal.

 

Acute Oral Toxicity Study

The acute oral toxicity procedure was followed by using OECD 423 guidelines starting with 2000 mg/kg body in adult female mice which were fasted overnight with water ad libitum. The starting dose level of the hexane extract of Citrullus lanatus was 2000mg/kg body weight p.o. as most of the crude extracts posses LD50 value more than 2000 mg/kg, p.o. Food was with held for further 3-4 hours after administration (p.o) of drugs and observed for the signs of toxicity. Body weight of the animals before and after administration were noted and any changes in skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous system and loco motor activity and behaviour pattern were observed, and also signed of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma were noted. The onset of toxicity and signs were also noted.

 

Evaluation for Anti-depressant activities

Experimental Design:

Animal: Mice, Sex: Male, Weight: 25-35 gm

 

On the day of the experiment, the animals were divided randomly into four groups of six animals each and treated with drugs for 1 weak.

Group I: Control (1% Tween 80; 10ml/kg, p.o)

Group II: EECL (250mg/kg in 1% Tween 80)

Group III: EECL (500mg/kg in 1% Tween 80)

Group IV: Standard (Imipramine; 10mg/kg, p.o in1% Tween 80)

 

Behavioural evaluation was carried out 60 minutes post drug/vehicle administration on the last day. The antidepressant activity of the test drug was evaluated using the following experimental models Forced Swim Test (FST) and Tail Suspension Test (TST) in mice.

 

Forced Swim Test (FST) in mice

Antidepressant activity was evaluated by using Porsolt Forced Swing test in mice.21,22 This works on the basic principle of antidepressant effect statistically decrease in immobility and behavioral despair in rodents. The Apparatus consist of a water tub of 60 cm (inner diameter) and 35 cm (height) was used. It was filled with water (27-29 °C) up to a height of 15 cm

 

All the animals were treated for one week with drugs. On day before experiment, each animal was dropped in water and was forced to swim for 6 min. It was then wiped dry and returned to home cage. On day 7th, mice were treated with drugs as mention in respective groups and control receive only vehicle. After a gap of 1 hour they were subjected to the swim test. In accordance with Porsolt et al, mice were kept in water for 6 min. The duration of immobility was recorded during the last 4 minutes of the observation period because each animal showed vigorous movement during initial 2 min period. The duration of the mouse was considered immobile when it floated motionlessly or made only those moments necessary to keep its head above the water surface. The water was changed after each test. The test was conducted in a dim lighted room and each mouse was used only once in the test.

 

Tail Suspension Test in Mice

In Tail Suspension test used is described by Steru, et. al. 22, 23 and works on the principle of Antidepressant effect significant decrease in escape oriented movement immobility (hangigg) in rodents.  The animals were hung by the tail on a plastic string 50 cm above the surface with the help of an adhesive tape, placed approximately 1 cm from the tip of the tail. Each animal under test was both acoustically and visually isolated from other animals during the test. The duration of immobility was observed for a period of 8 minutes. The duration of immobility was recorded during the last 6 minutes of the observation period. Mice were considered to be immobile only when they hung passively and were completely motionless. The test was conducted in a dim lighted room and each mouse was used only once in the test.

 

Evaluation of Anxiolytic and Skeletal Muscle Relaxant Activity

 

Experimental Design:

Animal: Mice, Sex: Male, Weight: 25-35 gm

 

On the day of the experiment, the animals were divided randomly into four groups of six animals each and treated with drugs for 1 weak.

Group I: Control (1% Tween 80; 10ml/kg, p.o)

Group II: EECL; (250mg/kg in 1% Tween 80)

Group III: EECL( 500mg/kg in 1% Tween 80)

Group IV: Standard (Diazepam; 10mg/kg, in 1% Tween 80, p.o)

 

Behavioural evaluation was carried out 60 minutes post drug/vehicle administration on the last day. The Anxiolytic activity of the test drugs were evaluated using the experimental Elevated Plus Maze (EPM) and Light-Dark Model (LDM) in mice.

 

 

Elevated plus Maze Model in Mice

Anxiolytic activity was accessed by using Elevated plus Maze(EPM) in mice .24 All the rodents have aversion for height and open space, they prefer to hide in enclosed arm therefore, spend greater amount of time in enclosed arm. Anxiolytic effect statistically increase in open time or open entries. The plus-maze apparatus, consisting of two open arms (16 x 5 cm) and two closed arms (16 x 5 x 12 cm) having an open roof. The drugs and vehicle were administered orally at their respective doses. After proper treatment with drugs each mouse was placed at the center of the maze with its head facing the open arm. During the 5 min experiment, the behavior of the mouse was recorded as the number of entries into the open or closed arms and time spent by the mouse in each of the arms. An arm entry was defined as the entry of all four paws into the arm.

 

Light-Dark Model in Mice

The light-dark model works on the principal that the light/bright environment works as source of anxiety and Anxiolytic effect statistically significant increase in light (movement) time or number of transition. 25-26 The mice’s light-dark box (40cm ×20cm ×20cm) consists of two parts, the light-compartment and the dark compartment. The box consists of a hole (5cm×5cm) in the bottom of the clapboard between the two compartments. The mice were treated with drugs and vehicles as respective groups and after one hour of treatment each mice during the test the mice were put into the center of the light compartment with their back to dark compartment and then transition behavior over 5 min was observed. Number of crossings between the light and dark area and total time spent in the illuminated part of the box were calculated. Every time before placing each animal, the maze was cleaned with 5% alcohol to eliminate the possible bias due the odor left by the previous animal.

 

Rota Rod test in Mice

The rotarod test is used to evaluate the activity of drugs interfering with motor coordination.27 The rotarod test was used to determine the effect of drugs on motor coordination. The instrument (a horizontal rotation device, Rota Rod, Edison) was set at a rate of 25 rpm. Each rat was placed on the rod and those animals that remained on the rod for 3 mins were selected for the study. The animals were than evaluated for motor coordination basal reading (the time each animal falls off from the rod/ time spent by animal in rotarod. After 60 min of administration of the tested drugs the each animal was kept on rotarod and the time each animal falls off from the rod each animal were recorded.

 

Grip strength test in Mice

In grip strength test, the mice was allowed to hold with the forepaws a steel wire (2mm in diameter and 35 cm in length), placed at a height of 50 cm over a cushion support.27 The length of time the rat was able to hold the wire was recorded. This latency to the grip loss is considered as an indirect measure of grip strength. After 60 min of administration of the tested drugs the each animal was kept on wire and the time each animal falls off from the wire of each animal were recorded.

 

Statistical Analysis

The data were expressed as mean ± standard error mean (SEM). The data were analyzed by using Graph pad software version5 by one way analysis of variance (ANOVA). The test was followed by Dennett’s‘t’-test, p values less than 0.05 were considered as significance.

 

RESULT:

Result of solvent Extraction

The coarse powder seeds were extracted with solvent extraction done n-hexane and oily light yellow colour extract was found. The percentage yield is 16.43% w/w.

 

Result of Preliminary Phytochemical Analysis

The hexane extract of Citrullus lanatus seeds  was tested for different phytoconsituents  and found to have Terpinoids, Flavanoids, Oils and Phenolic compounds.

 

Result of Acute oral toxicity

Acute oral toxicity studies of hexane extract of Citrulus lanatus (HECL) seeds were carried out according to OECD-423 guidelines in mice and starting at a dose of 2000 mg/kg, p.o. exhibited normal behaviour, without any signs of toxicity and two dose level as 250 mg/kg and 500 mg/kg selected for pharmacological activities.

 

Result of Evaluation of Anti-depressant activity

Effect of HECL on duration of immobility in Forced Swim Test in mice

The effect of pretreatment with HECL with (250 mg/kg, p.o and 500 mg/kg,p.o) for 7 days showed decreased in duration of immobility in Forced Swim Test in mice  and the result are given in Table-1  and plotted in Fig-1.  The effect on duration of immobility were compared with Standard (Imipramine; 10mg/kg, p.o) which showed significant decrease in duration of immobility.

 

Table-1: Effect of HECL on duration of immobility in Forced Swim Test (FST)

Group

Drug Treatment

Duration  of Immobility (sec.)  Mean ± SEM

I

Control

(1% Tween 80; 10ml/kg, p.o)

153.5±3.55

II

HECL

(250mg/kg in 1% Tween 80)

135.83±5.24*

III

HECL

(500mg/kg in 1% Tween 80)

127.8±3.00**

IV

Standard
(Imipramine; 10mg/kg,

p.o in 1% Tween 80)

74.67±3.82***

Values are in Mean ± S.E.M (n=6); ns -Non Significant, *p<0.05, **p<0.01, ***p<0.001 when compared with Control using One way ANOVA followed by Dunnetts multiple “t” test

 

 

Fig-1: Effect of HECL on duration of immobility in Forced Swim Test (FST

 

Table-2: Effect of HECL on duration of immobility in Tail Suspension Test (TST)

Group

Drug Treatment

Duration of Immobility (sec.) Mean ± SEM

I

Control

(1% Tween 80; 10ml/kg, p.o)

170.0±2.80

II

HECL

(250mg/kg in 1% Tween 80)

141.0±3.69*

III

HECL

(500mg/kg in 1% Tween 80)

130.0±4.62*

IV

Standard

(Imipramine; 10mg/kg, p.o in2% Tween 80)

72.33±2.69***

Values are in Mean ± S.E.M (n=6); ns -Non Significant, *p<0.05, **p<0.01, ***p<0.001 when compared with Control using One way ANOVA followed by Dunnetts multiple “t” test

 

Fig-2: Effect of HECL on duration of immobility in Tail Suspension Test (TST)

Effect of HECL for anxiolytic activity on EPM in mice

Effect of HECL on duration of immobility in Tail Suspension Test in mice

The effect of pretreatment with HECL with (250 mg/kg, p.o and 500 mg/kg,p.o) for 7 days showed decreased in duration of immobility in Tail suspension Test in mice  and the result are given in Table-2  and plotted in Fig-2.  The effect on duration of immobility were compared with Standard ( Imipramine; 10mg/kg, p.o) which showed significant decrease in duration of immobility.

 

The effect of pretreatment with HECL with (250 mg/kg, p.o and 500 mg/kg,p.o) for 7 days showed increase in no. of entries and time spent  in open arm of EPM in mice  and the result are given in Table-3 and plotted in Fig-3.  The effect on no. of entries and time spent in open arm of EPM were compared with Standard ( Diazepam; 10mg/kg, p.o) which showed significant increase in no. of entries and time spent  in open arm of EPM.

 

Fig-3: Effect of HECL for anxiolytic activity on EPM in mice


 

Table-3: Effect of HECL for anxiolytic activity on EPM in mice

Group

Treatment

No. of entries / 5min

Open arm    Close arm

Time spent (Sec)/5min

Open arm               Close arm

Group I

Control (Vehicle, 1% Tween 80; 10ml/kg, p.o)

4.50± 0.56

9.40±1.34

26.17±2.62

210.34±10.11

Group II

HECL (250mg/kg in 1% Tween 80)

6.33±0.24ns

8.45±0.78

61.83±6.23**

176.76±9.27

Group III

HECL (500mg/kg in 1% Tween 80)

6.50±0.76 ns

9.80±0.59

65.17±4.97**

164.67±8.45

Group IV

(Diazepam; 10mg/kg, in 1% Tween 80, p.o)

7.66±1.52**

8.45±1.39

129.7±6.66***

124.78±4.14

(Values are in Mean ± S.E.M (n=6); ns -Non Significant, *p<0.05, **p<0.01, ***p<0.001 when compared with Control using One way ANOVA followed by Dunnet’s “t” test.).

 


Effect of HECL for Anxiolytic Activity on Dark-Light Model

The effect of pretreatment with HECL with (250 mg/kg, p.o and 500 mg/kg,p.o) for 7 days showed increase in spent  in light chamber in Dark-light Model  in mice  and the result are given in Table-4 and plotted in Fig-4.  The effect on spent  in light chamber in Dark-light M0del  in mice  were compared with Standard ( Diazepam; 10mg/kg, p.o) which showed significant increase in   spent  in open chamber.

 

Table-4: Effect of HECL for Anxiolytic Activity on Dark-Light Model

Groups

Treatment

Time spent in Light-chamber

(Sec) Mean±SEM

Group I

Control (Vehicle, 1% Tween 80; 10ml/kg, p.o)

68.83 ± 4.62

Group II

HECL (250mg/kg in 1% Tween 80)

79.33 ± 4.28ns

Group IV

HECL (500mg/kg in 1% Tween 80)

88.0 ± 4.05*

Group V

 (Diazepam; 10mg/kg, in 1% Tween 80, p.o)

138.3 ± 5.35***

(Values are in Mean ± S.E.M (n=6); ns -Non Significant, *p<0.05, **p<0.01, ***p<0.001 when compared with Control using One way ANOVA followed by Dunnet’s “t” test.).

 

Fig-4: Effect of HECL  for Anxiolytic Activity on Dark-Light Model

 

Effect of HECL on skeletal muscle relaxant activity in Rota Rod Test in mice

The effect of pretreatment with HECL with (250 mg/kg, p.o and 500 mg/kg,p.o) for 7 days showed decrease  in spent  in rota rod  in mice  and the result are given in Table-5 and plotted in Fig-5.  The effect on time spent  rota rod  in mice  were compared with Standard (Diazepam; 10mg/kg, p.o) which showed significant decrease in time  spent  in rota rod.

 

 

Table-5: Effect of HECL on skeletal muscle relaxant activity in Rota Rod Test

Groups

Treatment

Time spent in Rota Rod

(Sec) Mean±SEM

Group I

Control (Vehicle, 1% Tween 80; 10ml/kg, p.o)

223.8± 5.48

Group II

HECL (250mg/kg in 1% Tween 80)

231.83± 6.04ns

Group III

HECL (500mg/kg in 1% Tween 80)

228.2± 8.29ns

Group IV

 (Diazepam; 10mg/kg, in 1% Tween 80, p.o)

21.17± 3.55***

(Values are in Mean ± S.E.M (n=6); ns -Non Significant, *p<0.05, **p<0.01, ***p<0.001 when compared with Control using One way ANOVA followed by Dunnet’s “t” test.).

 

Fig-5: Effect of HECL on skeletal muscle relaxant activity in Rota Rod Test

 

Effect of HECL on skeletal muscle relaxant activity on grip strength test

The effect of pretreatment with HECL with (250 mg/kg, p.o and 500 mg/kg,p.o) for 7 days showed decrease  in spent  in thread  in mice  and the result are given in Table-6 and plotted in Fig-6.  The effect on spent spent  in thread  in mice  were compared with Standard ( Diazepam; 10mg/kg, p.o) which showed significant decrease in  time spent  in thread.

 

Table-6: Effect of HECL on skeletal muscle relaxant activity on grip strength test

Groups

Treatment

Time spent in thread

(Sec)Mean±SEM

Group I

Control (Vehicle, 1% Tween 80; 10ml/kg, p.o)

73.17±5.51

Group II

HECL (250mg/kg in 1% Tween 80)

76.17±4.31ns

Group III

HECL (500mg/kg in 1% Tween 80)

75.00±4.26ns

Group IV

 (Diazepam; 10mg/kg, in 1% Tween 80, p.o)

17.33±1.45***

(Values are in Mean ± S.E.M (n=6); ns -Non Significant, *p<0.05, **p<0.01, ***p<0.001 when compared with Control using One way ANOVA followed by Dunnet’s “t” test.).

 

Fig-6: Effect of HECL on skeletal muscle relaxant activity on grip strength test

 

DISCUSSION:

Anxiety and depression form commonest stress-induced psychiatric disorders. To combat the biochemical changes which occur as a result of stress, there is antioxidant defence in the biological system. The role of anti-oxidant phytoconstituets widely screened for various diseases, such as cardiovascular disease, cancer, inflammation and allergy. But very less literature suggest the evidence for CNS diseases. A few studies available which suggest that antioxidant supplement therapy like vitamins A, C, and E  as an adjuvant therapy is useful in patients with stress-induced psychiatric disorders and the results have been discussed.28

 

In Forced Swim Test(FST) in mice, hexane exact of Citrulus lanatus (HECL) seeds showed dose dependent decreased duration of immobility but lower than standard imipramine. The decreased duration of immobility in Forced Swim Test reveals antidepressant activity.21   

 

Tail Suspension Test (TST) is a simple, rapid and reliable method to screen antidepressants and other class of psychotropics.  This method is based on the observation that a mouse suspended by the tail shows alternate agitation and immobility which is indicative of a state of depression. TST induced immobility is reduced by a large no of clinically active and atypical antidepressants.29 The hexane exact of Citrulus lanatus (HECL) seeds showed dose dependent decreased duration of immobility but lower than standard imipramine. The decreased duration of immobility in reveals antidepressant activity of the plant.

 

However depression is a complex disorder resulting from changes in central noradrenergic, serotonergic and dopaminergic systems or adrenomedullary system. Hence it was thought to be worthwhile to estimate all the three neurotransmitters in brain.

 

For evaluation of Anxiolytic activity, Elevation of Plus Maze apparatus was used. The elevated plus maze is a well-established animal model for testing anxiolytic drugs.30  It was found that dose dependent way the hexane exact of Citrulus lanatus (HECL) increases no. of entries and time spent in open arms but was less effective than standard diazepam.

 

In Light-Dark Transition test, the apparatus contains two compartments i.e. light and dark. Animals always try to spend more time in dark compartment because of fear about new environment. The light-dark test may be useful to predict the anxiolytic like activity of drugs in mice. It has the advantages of being quick and easy to use without food and water deprivation prior training of animals and natural stimuli are used. Transitions have been reported to be an index of activity exploration because of habituation over time and the time spent in each compartment to be a reflection of aversion.31  It was found that dose dependent way the hexane exact of Citrulus lanatus (HECL) increases no. of entries and time spent in light chamber but was less effective then standard diazepam.

In skeletal muscle relaxant activity, the pretreatment of hexane exact of Citrulus lanatus (HECL) seeds showed non significant activity in Rota rod as well as Grip strength test. Moreover, it was found to increase slight performance of the mice in both test models. The slight increase in performance is may be due to physical activity causing muscle fatigue due to release of ROS during activity, and anti-oxidants present in Citrulus lanatus scavenge those free radical. As some study already suggest that exercise and activity increases oxidative stress in cells 32 and antioxidants like alpha tocopherol increases swimming endurance of trained swimmers.33

 

CONCLUSION:

It can conclude that hexane extract of Citrullus lanatus Seeds possess anti-depressant and anxiolytic activity but does not affect on skeletal muscle activity. Finally, it can say that this can be used in CNS disorders like depression and anxiety and does not affect of skeletal muscle activity like synthetic drugs.

 

ACKNOWLEDGEMENT:

This study was supported by giving permission to use instruments and ethical clearance from institute. Our special thanks to Dr. M. Chinna Eswaraiah, Principal, Anurag Pharmacy College, Kodad, Andhra Pradesh, for his valuable suggestion and help.

 

REFERENCES:

1.       Rahman A.H.M.M., Anisuzzaman M., Ferdous Ahmed, Rafiul Islam A.K.M. and Naderuzzaman A.T.M.. Study of Nutritive Value and Medicinal Uses of Cultivated Cucurbits, Journal of Applied Sciences Research. 2008; 4(5): 555-558.

2.       Madhavi P, Maruthi Rao, Kamala Vakati, Habibur Rahman, M. Chinna Eswaraiah, Evaluation of Anti-Inflammatory Activity of Citrullus lanatus Seed Oil by In-vivo and In-vitro Models. Int. Res J Pharm. App Sci., 2012; 2(4):104-108.

3.       Alok Bhardwaj, Rajeev Kumar, Vivek Dabas,  Niyaz Alam, Evaluation of Anti-Ulcer Activity of Citrullus Lanatus Seed Extract in Wistar Albino Rats. International Journal of Pharmacy and Pharmaceutical Sciences. 2012; 4(5): 135-139.

4.       Okunrobo O. Lucky, Uwaya O. John, Imafidon E. Kate, Osarumwense O. Peter, Omorodion E. Jude, Quantitative determination, Metal analysis and Antiulcer evaluation of Methanol seeds extract of Citrullus lanatus Thunb (Cucurbitaceae) in Rats. Asian Pacific Journal of Tropical Disease. 2012; S1:261-S1265.

5.       Loiy Elsir Ahmed Hassan. In-vitro Antimicrobial activities of chloroformic, hexane and ethanolic extracts of Citrullus lanatus var. citroides. Journal of Medicinal Plants Research. 2011; 5 (8): 1338-1344.

6.       Swapnil Sharma. First report on Laxative activity of citrullus lanatus. Pharmacology online.  2011; 2:790-797. 

7.       Naresh Singh Gill. Evaluation of Antioxidant activity of citrullus lanatus Seed extract in Rats. Latin American journal of pharmacy (formely Acta Farmaceutica Bonaerense), Lat. Am. J. Pharm. 2011; 30(3):429-34.

8.       Madhavi P, Kamala Vakati, and Habibur Rahman. Hepatoprotective Activity of Citrullus Lanatus Seed Oil on CCl4 Induced Liver Damage in Rats.  Scholars Academic Journal of Pharmacy, 2012; 1 (1):30-33.

9.       Aruna Poduri, Debra L. Rateri, Shubin K. Saha, Sibu Saha, Alan Daugherty. Citrullus lanatus ‘sentinel’ (watermelon) extract reduces atherosclerosis in LDL receptor-deficient mice. Journal of Nutritional Biochemistry, 2012; (Article in Press).

10.     Ji Hyun Hwanga, Seong Gyu Ahna, Ju Youl Ohb, Young Whan Choia, Jum Soon Kanga, Young Hoon Park, Functional characterization of watermelon (Citrullus lanatus L.) EST–SSR by gel electrophoresis and high resolution melting analysis. Scientia Horticulturae. 2011; 130 :715–724

11.     Simonian N.A., Coyle J.T. Oxidative stress in neurodegenerative diseases. Annu. Rev. Pharmacol. Toxicol. 1996; 36: 83–106.

12.     Dhalla N.S,  Temsah R.M., Netticadan T. Role of oxidative stress in cardiovascular diseases. J. Hypertens. 2000; 18: 655–673.

13.     Bokov A,  Chaudhuri A, Richardson A. The role of oxidative damage and stress in aging. Mech. Ageing Dev. 2004,125:811–826.

14.     Rice-Evans C., Miller N., Paganga G. Antioxidant properties of phenoilc compounds. Trends Plant Sci. 1997; 2:152–159.

15.     Sealbert A., Johnson J., Saltmarsh M. Polyphenols: Antioxidants and beyond. Am. J. Clin. Nutr. 2005; 81: 2155–2175.

16.      Ross J.A., Kasum C.M. Dietary flavonoids: Bioavailability, metabolic effects, and safety. Ann. Rev. Nutr. 2002; 22: 19–34.

17.     Rice-Evans C.A., Miller N. Maxwell SJ. Prospects for the use of antioxidant therapies. Drugs. 1995; 49: 345.

18.     Rahman A.H.M.M., Anisuzzaman M., Ferdous Ahmed, Rafiul Islam A.K.M. and Naderuzzaman A.T.M.. Study of Nutritive Value and Medicinal Uses of Cultivated Cucurbits, Journal of Applied Sciences Research. 2008; 4(5): 555-558.

19.     El-Adawy T.A, Taha K.M. Characteristics and Composition of Watermelon, Pumpkin, and Paprika Seed Oils and Flours. J. Agric. Food Chem. 2001; 49(3): 1253-1259.

20.     Kokate CK, Practical Pharmacognosy, 5th Edn, Vallabh Prakasham, 107-121, 1991.

21.     Porsolt, R. D., Le Pichon, M., and Jalfre, M.  Depression: A new animal model sensitive to antidepressant treatments. Nature. 1977;266(5604): 730–732.

22.     Willner, P. Animal models of depression: an overview. Pharmacol Ther.1990; 45: 425-455.

23.     Steru L, Chermat R, Thierry B, Simon P. The tail suspension test: A new method for screening antidepressants in mice. Psychopharmacology (Berl.). 1985;85:367–370.

24.     Pelow S, Chopin P. File S.E. and BrileyM. Validation of openclosedarm entries in an elevated plus-maze as a measure of anxiety in the rat. J. Neurosci. Methods. 1985; 14 :149-167.

25.     Michel Bourin, Martine Hascoet. The mouse light/dark box test. European Journal of Pharmacology. 2003;463:55– 65.

26.     Ankit kumar Arora , M. Ashok, M. Veera Jyothsna, B. Radhakrishna, K.P. ShivalingeGowda. Evaluation of anxiolytic activity of aqueous and alcoholic extracts of leaves of Crataegus oxycantha in mice. Int J Pharm Biomed Sci. 2011;2(3): 86-91.

27.     Simiand J, Keane PE, Biziere K, Soubrie P. Comparative study in mice of tetrazepam and other centrally active skeletal muscle relaxants. Arch Int Pharmacodyn Ther. 1989 ;297:272-85.

28.     Medhavi Gautam, Mukta Agrawal, Manaswi Gautam, Praveen Sharma, Anita Sharma Gautam, and Shiv Gautam. Role of antioxidants in generalised anxiety disorder and depression. Indian J Psychiatry. 2012 ; 54(3): 244–247.

29.     Wesołowska A, Nikiforuk A, Stachowicz K and Tatarczyńska E. Effect of `the selective 5-HT7 receptor antagonist SB 269970 in animal models of anxiety and depression. Neuropharmacology. 2006;51(3):578-586.

30.     Kulkarni SK, Reddy DS. Animal behavioral models for testing antianxiety agents. Meth Find Exp Clin Pharmacol. 1996; 18: 219-230.

31.     Belzung C, Misslin R, Vogel E, Dodd RH, Chapounthier G. Anxiogenic effects of methyl-? carboline-carboxylate in a light-dark choice situation. Pharmacol Biochem Behav. 1987; 28:29-33.

32.     Powers SK, Jackson MJ. Exercise-induced oxidative stress: cellular mechanisms and impact on muscle force production. Physiol Rev. 2008;88(4):1243-76.

33.     Lawrence JD, Bower RC, Riehl WP, Smith JL. Effects of alpha-tocopherol acetate on the swimming endurance of trained swimmers. Am J Clin Nutr. 1975;28(3):205-8

 

 

 

Received on 03.05.2013                Modified on 10.06.2013

Accepted on 15.06.2013                © RJPT All right reserved

Research J. Pharm. and Tech. 6(8): August 2013; Page 878-884