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Ethnobotanical
Leaflets 13: 956-61, 2009. Preliminary
Phytochemical Screening and Evaluation of Anti-Inflammatory Potential of Vitex leucoxylon Linn. Gopalakrishna B1, Shukla
Prabodh2 and Shukla Padmini2 1R. R.
College of Pharmacy, Chikbanawara, 2Pranveer
Singh Institutes of Technology, Corresponding Author- pdmnmishra@gmail.com
Issued
July 01, 2009 Abstract The anti-inflammatory activity of ethanolic and
aqueous extract of the bark of Vitex
leucoxylon Linn. was studied in albino wistar rats using the Carageenan
induced rat paw edema model. The ethanolic and aqueous extract of Vitex leucoxylon (500 mg/kg p.o.)
inhibited Carageenan induced rat paw edema. The extract was also studied for
its preliminary phytochemical screening and acute toxicity studies. The
result indicated that both the extract produced significant (P<0.001)
anti-inflammatory activity when compared with the standard drug Indomethacin
(10 mg/ kg p.o.) and untreated control. Phyto-chemical screening showed the
presence of various constituent like phenolic compounds, phytosterol,
protein, flavonoids, carbohydrate etc. Key Words: Anti- inflammatory,
Vitex-leucoxylon, Carageenan. Introduction
Vitex leucoxylon Linn.
(Verbenaceae) commonly known as Songarbhi (Marathi) an excellent herbal crude
drug in the nature which has composition of the various essential constituent
that are required for normal and good health of human. It is small to large
tree with a short thick trunk and a spreading crown and almost throughout the
Deccan peninsula of India upto an altitude 900 mt, it extends northwards upto
Jhansi and part of Bihar.(1,2) The trees are generally found on
the river bank, stream & ponds. The bark is used as astringent whereas
the roots are febrifuge and astringent. The leaves are smoked for relieving
headache and catarrh and also used for medicinal baths in fever and anemia.
(3, 4) The purpose of the present study was to evaluate the anti-inflammatory potential of Vitex leucoxylon bark extract using carageenan induced rat paw model. The extract was also studied for its acute toxicity effects and preliminary phytochemical screening. Material and Methods Plant Material The bark of Vitex leucoxylon Linn. was collected
from the bank of pond known as Chennapur (Canara circle, Sirsi division) in
Bachagaon village, Chemicals Carageenan and Indomethacin was obtained from Preparation of Extracts For preliminary phytochemical screening alcoholic extract,
aqueous extract and successive extract (pet ether, ethyl acetate, alcoholic
and aqueous extract) of bark were taken. For the preparation of ethanolic extract of the bark of Vitex leucoxylon (EEVL) was collected,
shade dried at room temperature, pulverized and extracted with ethanol in a
soxhlet extractor for about 80 cycles. The extract was concentrated in a
rotary flash evaporator. The extract was divided into two portions one was
taken for screening (5, 6) and rest was suspended in Tween–80 for
anti-inflammatory activity. Similarly the aqueous extract (AEVL) was prepared
but water selected for extraction instead of alcohol. Here again the extract
was divided into two parts one was taken for screening and rest for
anti-inflammatory activity. For the preparation of successive extract the
dried drug was successively extracted from petroleum ether, ethyl acetate, alcohol
and in last with water. The solvents were evaporated in a rotary flash
evaporator and the dried residues were taken for screening. Animals Albino rats of wistar strain of the either sex (150-200gm)
maintained under standard environmental conditions (27±2º C light / dark
cycle of 12 hrs) and fed with standard pellet diet (Goldmohor brand, Lipton
India Ltd.) and water ad libitum
was used for the present study. All the experimental protocols were approved
by institutional Animal Ethics Committee. Toxicity Studies The 50% lethal
dose of the EEVL and AEVL was estimated by the up and down stair case method.
(7) Doses were administered orally at the dose of 100, 1000, 2000,
3000, 4000 and 5000 mg to six groups of animals. Control group received normal
saline (10 ml/kg) orally. Signs and mortality within 24-72 hrs were noted
CPCSEA Registration no.126/1999/CPCSEA. Anti-inflammatory activity The
anti-inflammatory activity of drug extract was assessed by the method
described by Winter et al. (8). The rats were divided into four
groups where six animals in each group were used for study. Acute
inflammation was produced by the sub-plantar administration of 0.1 ml of
1%w/v Carageenan ( %
inhibition = [1- Vt / Vc] X 100 Vc represented edema volume in control and Vt edema volume
in treated with test extracts. Statistical analysis – Results are expressed as Mean ±
SEM. The statistical analysis was performed by using unpaired student t- test
for comparing test groups with control group. P value less than 0.001 were
considered statistically significant. (9) Results Acute Toxicity Studies – In the acute toxicity test sign of
toxicity included lethargy, jerk, convulsion and death. No sign of toxicity
are seen at even the dose of 5 gm / kg. Preliminary Phytochemical screening
– As shown in
Table 1 preliminary phytochemical screening of different extract showed the
presence of Flavonoid, Carbohydrate, Phytosterol, Phenolic, Proteins and
glycosides. Anti-inflammatory activity- The result of EEVL and AEVL against
Carageenan induced paw edema is shown in Table – 2. The result showed that
the ethanolic extract of Vitex
leucoxylon exhibited significant activity at dose of 500 (mg/kg p.o.). As shown in table 2 aqueous extract of Vitex leucoxylon exhibited 46.06% inhibition in rat paw edema
whereas standard drug showed 50.6% inhibition of inflammation. Table 1. Preliminary Phytochemical analysis
of the bark of Vitex leucoxylon Linn.
Key: PE = Petroleum ether extract (60-800C) ALC = Alcoholic EA = Ethyl acetate AQ = Aqueous (+) = Present (-) = Absent Table 2. Effect of Indomethacin, EEVL and AEVL on
Carageenan induced paw edema in rats.
*Significant at p<0.001, P value
was calculated by comparing with control by ANOVA followed by Student T-
test, Values are expressed as Mean±SEM EV: edema volume EI:
Edema inhibition EV: edema volume EI:
Edema inhibition Discussion – Indigenous drug system can be
source of variety of new drugs which can provide relief in inflammation. The
most widely used primary test to screen new anti-inflammatory agent measure
the ability of a compound to reduce local edema induced in the rat paw
injection of a phlogistic agent. This edema depend on the participation of
kinins and polymorphonuclear leucocytes with there pro inflammatory factors
including prostaglandins. (10) The development of edema in the paw
of the rat after the injection of Carageenan has been described as a biphasic
event. The initial phase, observed around 1 hrs, is attributed to the release
of histamine and serotonin, the second accelerating phase of swelling is due
to the release of prostaglandin- like substances. It has been reported that
the second phase of edema is sensitive to both clinically useful steroidal
and nonsteroidal anti-inflammatory agents. (10, 11) Significant
anti-inflammatory activity was observed for ethanolic and aqueous extract of Vitex leucoxylon in Carageenan induced
edema model. Hence the anti-inflammatory activity in Vitex leucoxylon can be attributed to the presence of flavonoid
and tannins. So from the above study it is quite apparent that the
aqueous extract of Vitex leucoxylon
plants possesses significant anti-inflammatory activity. The further study
justifies its use in inflammation, pain and wound healing as suggested in the
folklore medicines. References 1.Nandkarni A.K., Indian Materia Medica, Popular Book
Depot. 2. The Ayurvedic Pharmacopoeia of 3.Database on medicinal Plants used in Ayurveda and Siddha,
Govt. of India, 4.The Wealth of 5.Kokate C.K., Purohit A.P., Gokhale S.B., Textbook of Pharmacognosy, Nirali Prakashan Pune, 36:
593-597. 6.Khandelwal K.R., Practical Pharmacognosy, Nirali
Prakashan, 7.Winter 8.S.M. Hess and R.C.Milonig, Assay for Anti-Inflammatory
Drugs. In: Lepaw H. Ward. PA, Editors. Inflammation, Mechanism and control. 9.Woodson R.F., Statistical methods for analysis of
biochemical data Wiley, 10.Damas J. Remacle-Volon G. and Deflandre E., Further
studies of the mechanism of counter irritation by Turpentine. Arch.
Pharmacol.332: 196-200 (1986). 11.Beatriz B., Gerardo M., Arrrtonio J.L., and Jose A.S.E.
Anti-Inflammatory activity Urera
baccifera (Urticaceae) in sprage dawley rats, Research paper available on
line: http:// rbt.ots.ac.cr |
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