INTRODUCTION:
Cyathocline lyrata is annual herb, growing to 20-25
cm hight, branched
grooved stem has soft hair covering it. Whole Plant is strongly aromatic.
Alternatively arrange stalkless leaves are toothed
covered with soft hair and flowers occurs in corymbs at the end of
branched in purple color. Cyathocline
lyrata widely spread in Himalyas
range, Assam, India
and also available in Local
area of Betul
M.P. Cyathocline lyrata is
well known drug
in Indigeneous system of medicine for its
various used as a bitter tonic. It acts
as germicide and appetizer. The
essential oil of aerial part of Cyathocline
lyrata had show fairly pharmacological
activity. It also shows anthelmatic, insect repellant and anti-microbial activity..
Inflammation is the complex biological response of vascular tissues to harmful
stimuli, such as pathogens, damaged cells or irritants. Inflammation is a protective attempt by the
organism to remove the injurious stimuli as well as initiate the healing
process for the tissue.
Non
steroidal anti-inflammatory drugs
(NSAID’s) are
widely used in the
treatment of pain,
fever and inflammation. However these drugs have no
side effects especially on the gastro intestinal tract.
Therefore there is a need to search for novel
anti-inflammatory agents from
natural source, which
could be used
in medicine and as
additives to Nutraceuticals.
Many natural products have been reported to have anti-inflammatory action.
An attempt is made
during this research work
to evaluate the
chloroform extracts of whole part of Cyathocline lyrata for anti-inflammatory activities.
MATERIAL AND METHOD:
Plant material collection and authentication:
The entire plant of Cyathocline lyrata
Cass were collected from local area of Betual
(M.P.) in the month of June, 2012. The plant specimen was confirmed
by Dr. Madhuri Modak,
Professor, Dept. of Botany, Motilal Vigyan Mahavidyalaya, Bhopal
(M.P). A voucher specimen (Her/Bot/1212.90-374) is
deposited at the herbarium of MJPR University, Jaipur
(Rajasthan).
Evaluation of In vivo anti-inflammatory activity:
Procurement of
Experimental Animals:
Male albino rats (100-150 g) of approximate same age were used in the
present studies was procured from VNS College of pharmacy, Bhopal India. The animals were fed with standard pellet
diet and water ad libitum. All the animals
were housed in polypropylene cages. The animals were kept under alternate cycle
of 12 hours of darkness and light. The animals were acclimatized to the laboratory condition for 1 week before starting the
experiment. The animals were fasted for at least 12 hours before the onset of
each activity. The animals received the drug treatments by oral gavage tube. The preparation of chloroform extract the next step was
to formulate a suspension of extracts of Cyathocline lyrata Cass which was subjected to
animal studies. Suspension of the extract was made by suspending in 0.5% CMC.
Acute toxicity studies:
This study is needful before pharmacological screening
on animals. The acute oral toxicity study of Cyathocline lyrata chloroform extracts was carried
out according to OECD 423 guideline (Organization for Economic Cooperation and
Development) which is based on a stepwise procedure with the use of a minimum
number of animals per step. All animals were received respective dose for seven
days and observed mortality on 7th day. The Animal Male albino rats (100-150 g)
were treated with the drug of chloroform extract Cyathocline lyrata.
Screening of Anti-inflammatory Activity:
Carrageenan
induced paw oedema[9]
Both in- vivo and in- vitro methods are available for
the evaluation of anti-inflammatory agent but among the in-vivo method the carrageenan induced rat paw oedema
assay is belived
to be one of the most reliable and also the most widely used.
Male Swiss albino rats weighing 100- 150 gm
were used. The animals were put on standard diet and water was provided ad libitum.
The animals were fasted over night before the experimentation. The rats were
divided into six groups (n=6). The
anti-inflammatory activity of extract was assessed by the method described by
Winter et al. Rats were divided into four groups
Group I Normal control, without treatment,
Group II:
Negative
control, intraplantar injection of 0.1 ml of 1 %( w/v) carrageenan on 7th Day.
Group III:
Positive
control, received Diclofenac potassium 10mg/kg BW p.o.
on 7th day before 1hr prior to intraplantar injection of 0.1 ml of carrageenan
Group IV:
Rats received chloroform
extract of Cyathocline lyrata
(100-200 mg/kg) for 7 days p.o.
and then after 1hr of last dose, intraplantar
injection of 0.1 ml of carrageenan.
Group V: Rats chloroform extract of Cyathocline lyrata (200-400 mg/kg) for 7
days p.o. and then after 1hr of last dose, intraplantar injection of 0.1 ml of carrageenan
Since the LD50 has not been determined
during the acute toxicity study, the doses for this study where selected by
trial and error method. The standard and extract were given orally to the
animal. The perimeter of paw was measured by using vernier
caliper mesurments were taken at 0,1,2,3 hours after
the administration of the carrageenan.
The
percentage inhibition of edema was calculated by the following equation:
% Inhibition
of edema= 100 (
)
Where Vc is the edema volume in the control group and Vt is the edema volume in tested groups [Il-Ok
Lee, You-Seong Jeong, 2002].
Formalin
induced paw edema [10]
In the
experiment, a total of 24 rats were used. The rats were divided into 5 groups
Comprising of 6
animals in each group as follows:
Group I: Normal control, without treatment,
Group II: Negative control, intraplantar injection of 0.1 ml of formalin on 7th
day.
Group III Positive control, received
Diclofenac potassium 10mg/kg BW p.o. on 7th
day before 1hr of intraplantar injection of 0.1 ml of formalin
Group IV Rats received chloroform extract of Cyathocline lyrata
(100-200 mg/kg) for 7 days p.o.
and then after 1hr of last dose, intraplantar
injection of 0.1 ml of formalin.
Group V Rats chloroform extract of Cyathocline lyrata
(200-400 mg/kg) for 7 days p.o.
and then after 1hr of last dose, intraplantar
injection of 0.1 ml of formalin.
Acute
inflammation was caused by injecting 0.1 ml of formalin (40%) into the
sub-plantar region of the right hind paw of each rat. The paw volume was
measured plethysmometrically at 0 h, 1 h, 2 h, 3 h,
4h, 5h and 6h after the formalin injection. Edema was expressed as mean
increase in paw volume relative to control animals. The percentage inhibition
of edema was calculated by the following equation:
% Inhibition
of edema= 100 (),
Where Vc is the edema volume in the control group and Vt is the edema volume in tested groups