ANTIANXIETY ACTIVITY OF EMBELIN ISOLATED FROM EMBELIA RIBES

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About Authors:
Mukesh Bansal1*, Indrajeet Singhvi2, Santosh Gupta2
1Department of Pharmaceutics, Indian Institute of Technology (Banaras Hindu University), Varanasi, U.P., India.
2Pacific College of Pharmacy, Pacific University, Udaipur, Rajasthan, India.
*bansalpharma1987@gmail.com

Abstract:
Object:
To investigate the effect of embelin  on anxiety   was studied  by using standard test such as Elevated zero-maze test and Novelty induced suppressed feeding latency test in a dose dependent manner.
Material & methods:
This portion include extraction and isolation of embelin, animals were obtained from the animal house IMS, BHU, Varanasi, India, and methods for anxiety were Elevated zero-maze test and Novelty induced suppressed feeding latency test.
Result & conclusion:
The  anxiolytic  activity was found at 20mg/kg. The observed anxiolytic effect of this compound seems to be regulated through monoaminergic system in the brain and the activity was comparable to diazepam.

Reference Id: PHARMATUTOR-ART-1568

Introduction
Embelin (2,5-dihydroxy-3-undecyl-1,4-benzoquinone) is a major constituent of Embelia ribes   (Family: Myrsinaceae). The whole plant is used as anti-inflammatory to relieve rheumatism and fever (1). The fruit is good appetizer, cures tumors, ascites, bronchitis, jaundice, brain tonic, mental disorders, dyspnaea, diseases of the heart; urinary discharges, scorpionsting, snake-bite and tooth ache (2). Embelin showed antifertility, antitumor, anti-nflammatory, analgesic, antioxidant, hepatoprotective, wound healing and antibacterial (3,4,5) activities. Incidence of anxiety in developed countries is approximately 20 per 100,000 while that of developing countries is 50 per 100,000. The entire currently available antianxiety drugs are synthetic molecules associated with side effects and approximately 30% of the patients continue to have anxiety with this therapy (6) on withdrawl. In anxiety majority of evidence supports the brain deficiency of serotoninergic and dopaminergic activity, while over activation of noradrenergic system occurs (7). Till date no comprehensive report is available on Anxiety profile of Embelin. So  In the present study, we investigated the antianxiety activity of embelin by using Elevated zero-maze test and Novelty induced suppressed feeding latency test.

Materials and methods

Extraction and isolation of embelin
The berries of Embelia ribes were purchased from Abirami Botanicals, Tuticorin, TN, India and authenticated by Medicinal Plants Survey and Collection Unit, Department of biology, Banaras Hindu University,Varanasi, India. Coarsely powdered berries (1 kg) were extracted for 72 h with n-hexane by cold maceration (3×2 l). The extract was concentrated to dryness in a rotavapor and chromatographed over silica gel (100–200 mesh). Elution with benzene and re-crystallization with ether afforded orange plates of embelin (yield 3 g, 0.3%, (8). It was found to be 100% pure by HPTLC using the solvent system ethyl acetate: benzene (70:30) and characterized by comparing its physical and spectral values with literature (9).

Animals
The animals were obtained from the animal house IMS, BHU, Varanasi, India, maintained under standard conditions (12 h light/dark cycle; 25±3 ?C, 45–65% humidity), had free access to standard rat feed and water ad libitum and studies were performed. Chemicals Diazepam (Ranbaxy, New Delhi, India),   purchased.All other chemicals used were of analytical grade.

Elevated zero-maze test
Rats were placed on one of the enclosed quadrants for a 5 min test period. The maze was cleaned with 5% ethanol/water solution and dried thoroughly between test sessions. During the 5 min test period, time spent on open arms, number of `head dips’ over the edges of platform, and number of `stretched attend postures’ from closed to open quadrants were recorded (10,11).

Novelty induced suppressed feeding latency test:
Naive rats were placed individually in the test chamber and the latency to begin eating (defined as chewing of the pellet and not merely sniffing or playing with it), were recorded.  If the rat had not eaten within 300 sec, the test were terminated and latency score 300 sec was assigned. A neutral ‘blind’ observer made observations (12).

RESULT:

Anxiolytic activity
Elevated zero maze tests:

Significant fall (p<0.05) in the entries in open arms, duration in open arms no. of head dips, stretched attend postures was observed in the control animals with stress compared to control without stress. Significant increase in the entries in open arms (p<0.05), duration in open arms (p<0.05), no. of head dips (p<0.05), stretched attend postures (p<0.05) with subsequent increase in the dose of embeline was observed which proves that embeline and diazepam have beneficial effect on chronic stress induced anxiety.The results are described in Table 1.

Table 1: Anxiolytic activity of elevated zero maze tests of the embeline

Treatment

Dose

(mg/kg, p.o.)

Time spent in open arms (sec)

Entries in open arms (N)

Head dips

(N)

Stretched attend postures

(N)

Control

Distilled water

54.53±1.50

5.33±0.33

8.17±0.17

3.27±0.17

Control + Stress

Distilled water

32.50±0.56*

3.00±0.26*

5.00±0.35*

2.00±0.26*

DNMB+ Stress

5

32.67±0.71#

2.96±0.11#

5.27±0.23#

2.46±0.28

DNMB + Stress

10

30.43±0.38#

4.00±0.47#

6.67±0.43#

4.37±0.16#

DNMB + Stress

20

50.27±0.47#

7.50±0.33#

11.50±0.37#

4.00.71#

Diazepam  + Stress

5

51.17±0.64#

8.35±0.24#

13.39±0.28#

4.23±0.12#

Values are mean ±SEM, n=6. Statistical analysis by one way ANOVA followed by Dunnet’s multiple comparison test; *P<0.05 compared with Control; #P<0.05 compared with Control + stress.

Novelty induced suppressed feeding latency test:
Significant rise (p<0.05) in the time of latency to feed was observed in the control animals with stress compared to control without stress. Significant fall (p<0.05) in the time of latency to feed with subsequent increase in the dose of embeline was observed which proves that embeline and Diazepam have beneficial effect on chronic stress induced anxiety.The results are described in Table 3.

Table 3: Anxiolytic activity ofNovelty induced suppressed feeding latency of embeline

Treatment

Dose

(mg/kg, p.o.)

Time of latency to feed (sec)

Control


Distilled water

26.50±2.42

Control + Stress


Distilled water

23.50±0.16

DNMB+ Stress


5

21.07±1.11

DNMB + Stress


10

30.63±0.78

 DNMB + Stress


20

45.27±0.86

Diazepam  + Stress


5

50.17±1.44

Discussion:
Although there are so many application of embelia ribes such as antifertility, antitumor, anti-inflammatory, analgesic, antioxidant, hepatoprotective, wound healing and antibacterialknown  have been since centuriesbut now embelin was isolated rom plant embelia ribes . However therapeutically interesting pharmacological activity profile in animal models of any given drug can be quantified only by the use of its appropriate dose range and treatment regimen.  Moreover, during literature survey no reports on antianxiety activity of the embelin could be identified. The result indicates that increase in  time spent in open arms (sec) and increase in number of head dips (N)having the evidence of antianxiety activity in Elevated zero maze test & inNovelty induced suppressed feeding latency test increase in time of latency to feed (sec) at 20 mg/kg indicate antianxiety which was comparable with diazepam at 5 mg/kg. These effects of this compound were not  strictly dependent on its dose because at 5 & 10mg/kg embelindid not show better effect than 20mg/kg . These observed anxiolytic effect of embelinseems to be regulated through monoaminergic system in the brain. In conclusion, embelin exhibited significant activity in both model Elevated zero maze test &Novelty induced suppressed feeding latency test at 20mg/kg.

References
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