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WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES Thomas et al.

World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 5.210

Volume 4, Issue 08, 1245-1253.

Research Article

ISSN 2278 – 4357

EVALUATION OF ANXIOLTIC EFFECT OF WHOLE PLANT OF “CUSCUTA REFLEXA” Sujith Thomas*1, Sapna Shrikumar2, C Velmurugan3 and B.S. Ashok Kumar4 1 2

Department of Pharmacology, Moulana College of Pharmacy, Kerala-679321.

Department of Pharmacognosy, Moulana College of Pharmacy, Kerala-679321. 3

Department of Pharmacology, Sri Krishna Chaithanya College of Pharmacy, Madanapalle, AP-517325

4

Department of Pharmacognosy, Sri K.V. College of Pharmacy, Chickballapur, Karnataka.

Article Received on 27 May 2015, Revised on 22 June 2015, Accepted on 15 July 2015

ABSTRACT The anxiolytic effect of methanol extract of whole plant of Cuscuta reflexa at doses of 200 and 400 mg/kg was evaluated by elevated plus maze and light and dark chamber in mice. Animals were treated orally with normal saline, extract and diazepam 5 mg/kg used as standard

*Correspondence for Author

drug. The extract 400 mg /kg showed significant anxiolytic effect compared to 200 mg/kg in both models. The 400 mg/kg effect was

Sujith Thomas Department of

comparable to standard. Thus methanol extract of Cuscuta reflexa

Pharmacology, Moulana

could serve as good anxiolytic agents and seems to be promising for

College of Pharmacy,

the development of phytomedicines for anxiety.

Kerala-679321.

KEYWORDS: Anxiolytic, Cuscuta reflexa, methanol, light and dark chamber, elevated plus maze. INTRODUCTION The science and technology have contributed to an enormous improvement in the quality of life of humankind. However, modern life stress and associated trails and tribulation are responsible for the surge in incidence of variety of psychiatric disorders. Anxiety disorders, the most prevalent psychiatric illnesses in the general community are present in 15 to 20 percent of medical clinic patients. The primary anxiety disorders are classified according to their duration and course and the existence and nature of precipitants.[1] Approximately onethird of medical patients presenting with anxiety have an organic etiology for their psychiatric symptoms, but an anxiety disorder can also present with somatic symptoms in www.wjpps.com

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absence of diagnosable medical condition.[2] Path breaking research in psychopharmacology has floded the market place with drugs for specification. For instance, benzodiazepines (diazepam, nitrazepam, lorazepam and alprazolam etc.) are the most frequently prescribed synthetic drugs for variety of condition particularly anxiety, depression, epilepsy and insomnia. But these psychoneural drugs have very serious side effects like chronic use of benzodiazepines causes detioriation of cognitive function, physical dependence and tolerance. Besides addiction-liabilities, benzodiazepines adversely affect the respiratory, digestive and immune system of body. Since the chronic treatment with benzodiazepines often prove more harmful in the longer run.[3] Cuscuta reflexa Roxb. is a rootless, leafless perennial parasitic twining herb of Convolvulaceae family, commonly known as Akashvalli or Dodder. The plant is distributed worldwide and in India about 6 species are found. It has no chlorophyll and cannot make its own food by photosynthesis. It grows on thorny or other shrubs, sometimes completely covering the bushes and trees.[4] The Cuscuta reflexa is investigated for antitumor[4], antimicrobial[5], hepatoprotective[6], anticonvulsant[7], antioxidant[8], induced alopecia[9] activities. Many chemical constituents have been isolated from Cuscuta reflexa such as cuscutin, amarbelin, beta-sterol, stigmasterol, myricetin, qurecetin, cuscutamine, luteolin, bergenin[10]etc. So this is medicinal important plant having numerous pharmacological activities. So the present study designed to evaluate the anxiolytic effect of whole plant of Cuscuta reflexa. MATERIAL AND METHODS Collection and authentication of the plant material The whole plant of Cuscuta reflexa had been collected from the medicinal garden of sri Krishna Chaithanya college of Pharmacy, Chittoor District, Andhra pradesh, India. The plant was identified and authenticated by the Botanist Dr. K. Madhava Chetty, Assistant Professor, Department of botany, Sri Venkateswara University, Tirupathi. Extraction procedure The fresh whole plant of Cuscuta reflexa were collected and dried under shade and ground into powder with mechanical grinder. The powder was passed through sieve no. 40. The 250gm of dried powder of Cuscuta reflexa was defatted with petroleum ether. The defatted powder material (marc) thus obtained was extracted with methanol by maceration.[11]

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World Journal of Pharmacy and Pharmaceutical Sciences

Table: Data showing the % yield of dried powder of Cuscuta reflexa Plant name

Part used

Cuscuta reflexa

Whole plant

Method of extraction Maceration

Colour of extract greenish Brown

Nature of extract Semi solid

% yield of extract 12.56%

Preliminary Phytochemical Investigation Methanol extract of Cuscuta reflexa was subjected to qualitative analysis for the various phytoconstituents.[12] Acute oral toxicity study[13] Acute oral toxicity studies was performed as per OECD-423 guidelines (acute toxic class method), with methanol extract of Cuscuta reflexa. Swiss albino mice with weight ranging (20-25 gm female) were taken for the experiment. The animals were made into a group of third each, dose of methanol extract was given according to the body weight (mg/kg), starting dose of 5 mg /kg was given to the first individual animal, no death was occurred, and higher doses were given to next group of animals. Table: Acute toxicity study of Methanol extract of Cuscuta reflexa based on OECD 423 guidelines. S. No 1 2 3 4

Number of animals 3 3 3 3

Dose in mg/kg 5mg/kg 50mg/kg 500mg/kg 2000mg/kg

Report No death No death No death No death

Table: Study period and observation parameters Initial once observation Special attention Long term observation

Direct observation parameters

Additional observation parameters

First 30 min and periodically 24 hr First 1-4 hr after drug administration Up to 14 days Tremors, convulsions, salivation, diarrhea, lethargy, sleep and coma. Skin and fur, eyes and mucous membrane, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavior pattern etc.

From the observation the methanol extract of Cuscuta reflexa was does not cause any mortality and morbidity up to 2000mg/kg. Hence 2000mg/kg consider as LD50. Therefore its www.wjpps.com

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ED50 was found to be 1/10th of LD50 as 200 mg/kg. The present study doses was fixed by ED50 as 200 & double the dose of ED50 as 400 mg/kg of whole plant of methanol extract of Cuscuta reflexa ANTI-ANXIETY ACTIVITY Experimental design Swiss albino mice (20-25gm) were taken and divided into four groups, each group comprised of 6 animals. Group I

: Negative control, administer normal saline as vehicle orally.

Group II

: Received Cuscuta reflexa 200 mg/kg orally.

Group III

: Received Cuscuta reflexa 400 mg/kg orally.

Group IV

: Received standard drug Diazepam 5mg/kg orally.

Elevated plus maze[14] The apparatus consist of two open arms (5x10cm) and two closed arms (5x10x15cm) radiating from a platform (5x5cm) to form a plus-sign figure. The apparatus was situated 40 cm above the floor. The open-arm edges were 0.5 cm in height to keep the mice from falling, and the closed-arm edges were 15 cm in height. Drugs and vehicle were administered to the animals 30 min prior to study as per experimental design. Then the animal was placed at the center of the maze, facing one of the closed arms and observed for 5 min. During 5 min test period the number of entries into open and closed arms and time spent in the open arms and closed arms were noted. Arm entry was counted when the animal has placed all of its four paws on it. The procedure was conducted preferably in a sound attenuated room, with observations made from an adjacent room. Light and Dark chamber[15] The apparatus consisted of a light and dark unit, consisting of a box with two distinct chambers, black and white, connected by a small open door way. The dark compartment (15x20cm) had its sidewalls and floor covered by a smooth black colored sunmica material. The bright compartment (20x20cm) had its side walls and floor covered by white colored sunmica material; it was brightly illuminated with a 60 W milky white bulb located 28 cm above the floor on the partition wall. An opening of (7x9cm) was located at a floor level in the center of the partition to allow access between two compartments for the animal. Drugs and vehicle were administered to the animals 30 min prior to study as per experimental design. At the start of the test, a mouse is placed in the middle of the illuminated part of the www.wjpps.com

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cage. The number of crossing into light and dark chamber is counted and time spent in light and dark chamber during 10 min period were noted. STATISTICAL ANALYSIS Results were expressed as Mean±SEM .The differences between experimental groups were compared by one-way Analysis of Variance (ANOVA) followed by Dunnett’s test and were considered statistically significant when *p