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Jolly John et al., J. Pharm. Res. 2014, 3(2), 11-13
Journal of Pharma Research 2014, 3(2) 11-13
J
ournal of
P
harma
R
esearch
Research Article
Available online through ISSN: 2319-5622
www.jprinfo.com
Anti-inflammatory Effect of Quassia indica leaf extract And Role of Antioxidant activity
Jolly John*, Suraj.S, Abdul Vahab A, Dr Jyoti Harindran, S.E Godwin
Department of pharmacology, Regional institute of medical science and research centre, UCME puthupally, kottayam, kerala.
Received on: 22-02-2014; Revised and Accepted on: 01-03-2014
ABSTRACT
T
he plant Quassia indica belongs to the family Simarubiaceae is found mainly in India and Srilanka. This plant has been widely used in
traditional system of medicine in India. The in-vitro antioxidant activity of methanolic extract of Quassia indica leaves were estimated by DPPH
radical scavenging method. Anti inflammatory activity of Quassia indica leaves was tested against carrageenan induced paw edema in wistar
albino rats. Here diclofenac used as standard drug for comparison. The study reveals that it possesses significant anti-inflammatory and
antioxidant activity.
Key words: Quassia indica, Antioxidant, Anti-inflammatory,
carageenan, diclofenac.
INTRODUCTION
H
erbal drugs constitute a major part of therapeutics in
all the traditional systems of medicine. There are approximately
1250 Indian medicinal plants, which are used in formulating
therapeutic preparations according to Ayurveda and other
traditional systems of medicine. Free radicals can also react with
DNA, proteins or lipids in the cell membrane and cause cellular
damage which is the root cause of many diseases. Flavonoids are
phytochemicals known to possess anti-inflammatory, antioxidant,
antiallergic, hepatoprotective, antithrombotic, neuroprotective,
anticancer activities. Quassia indica (simarubiaceae) is one such
bitter plant which is abundantly grown and used as important
medicinal plant. However much of its medicinal importance is not
assessed. By literature survey it is found that the leaves of the plant
contain main phytochemical constituents such as quassinoids,
samaderin b, samaderin c. Earlier studies shows that its related
species shows anti-oxidant and anti-inflammatory activity, due to
the presence of flavanoids
[1-5]
.
MATERIALS AND METHOD
T
he fresh leaves of Quassia indica was collected from the
locally growing area of Allapuzha, Kerala in February 2012. The
plant was identified and authenticated by Dr. K.V.George,
Department of Botany, C.M.S College Kottayam, Kerala. A herbarium
specimen is deposited in our college museum.
(UCP/MGU/RIMSR/2012/herb9).
Extraction:
The leaves were shade dried and coarsely powderd; 100
gm of powder were soaked in 500ml of methanol for 3-5 days with
intermittent shaking. At the end of the extraction, it was passed
through whattman filter paper. This filtrate was concentrated under
reduced pressure on rotary evaporator .The yield was 16.4 % w/w
and subjected to phytochemical screening to identify the various
phytoconstituents.
Experimental animals:
Female rats of thirty healthy weighing 150-200gms and
three female albino mice weighing between20-25gms were
*Corresponding author:
Jolly John
Department of pharmacology,
Regional institute of medical science and research centre,
UCME puthupally, kottayam, kerala.
Ph. No: 8547679527
.
*E-Mail: jollyjohn03@gmail.com
obtained from the animal house of UCP, Regionial institute of
medical sciences and housed in polycarbonate cages. The animals
were housed under standard conditions. Approved at the
Institutional Animal Ethics Committee (IAEC) of UCP, RIMSR was
taken for conducting Anti-inflammatory activity.
Acute toxicity study:
Acute toxicity study was performed to ascertain safe dose
by Organization of Economic Cooperation and Development (OECD)
423 guidelines
[6]
.
Antioxidant activity of Quassia indica leaf extract: Invitro
DPPH free radical scavenging assay:
The free radical scavenging capacity of the extracts was
determined using DPPH. DPPH solution (0.004% w/v) was prepared
in 95% methanol. Methanolic extract of Quassia indica was mixed
with 95% methanol to prepare the stock solution. DPPH solution
was taken in test tubes and Quassia indica extracts was added
followed by serial dilutions from100 μg to 500 μg to every test tube,
made up to final volume 3 ml, the absorbance was read at 515 nm
using a spectrophotometer after 10 min. Ascorbic acid was used as a
reference standard which dissolved in distilled water to make the
stock solution with in the same concentration 5mg/ml
[7, 8]
.
% scavenging of the DPPH free radical was measured using the
following equation = (Absorbance of the control - absorbance of the
test sample) / (absorbance of the control) *100
Anti-inflammatory activity by Carrageenan induced paw edema
in rats: Invivo:
Experimental procedures:
Wistar albino rats weighed around 150 to 250 were used
for this study. The initial right hind paw volume of the rats was
measured using a plethysmometer. They were divided into 5
groups consists of 6 rats each.
Group-1: Negative control which received vehicle distilled water
only.
Group-2: Positive control which received carrageenan 0.1ml 1%
intra plantar.
Group-3: Standard which received only diclofenac sodium 5mg/kg
p.o+ carrageenan 0.1ml 1% intra plantar.
Group-4: Test group which has received the extract 200 mg/kg p.o+
carrageenan 0.1ml 1% intra plantar.
Group-5: Test group which has received the extract 400 mg/kg p.o+
carrageenan 0.1ml 1% intra plantar.
Jolly John et al., J. Pharm. Res. 2014, 3(2), 11-13
Journal of Pharma Research 2014, 3(2) 11-13
Procedure:
Anti-inflammatory activity was assessed by test group’s
methanolic leaf extact 200,400 mg/kg and negative control vehicle,
standard group diclofenac sodium was given orally. After 30 min,
the rats were challenged with intra plantar injection of 0.1 ml of 1%
w/v solution of carrageenan into the sub plantar region of paw. The
paw was marked with ink at the level of lateral malleolus. The paw
volume was measured at 1, 3, 6 hours after carrageenan injection
using a Plethysmograph. The difference between initial and
subsequent reading given the actual edema volume. The anti-
inflammatory activity in animals that received Quassia indica
extracts and diclofenac (5mg/kg) was compared with that of vehicle
control groups
[9, 10]
.
Statistical analysis:
Results were expressed as mean ± SEM, (n=6). Statistical
analysis were performed with one way analysis of variance
(ANOVA) followed by Dunnett’s t test. *P<0.05, **<0.01 and
***<0.001, show statistical significance when compared treatment
group with control group.
RESULTS AND DISCUSSION
T
he preliminary phytochemical analysis revealed the
presence of alkaloids, tannins and phenolic compounds, triterpenes,
carbohydrate, flavanoids in methanolic leaf extract. Acute toxicity
studies for methanolic leaf extracts of Quassia indica belonging to
the family Simarubiaceae were conducted as per OECD guidelines
423 using albino Swiss mice. The extracts were found to be safe up
to 2000 mg/kg body weight since no death and signs of toxicity
were observed.
Free radicals involved in the process of lipid peroxidation
are considered to play a cardinal role in numerous chronic
pathologies, such as cancer and cardiovascular diseases among
others, and are implicated in the ageing process
.
Methanolic leaf
extract of Quassia indica have demonstrated dose dependent
increase in the DPPH radical scavenging activity. Table. 1. The IC50
value of MEQI extract was found to be 190μg/ml. Therefore, the
extract were assessed against scavenging of free radicals like DPPH
and compared against standard shows good antioxidant activity
Fig.1.
Inflammation is part of the complex biological response of
vascular tissues to harmful stimuli such as pathogens, damaged
cells, or irritants. In Carrageenan-induced rat paw edema, the
injection of carrageenan, the phlogistic agent, caused localized
edema starting at 1 hour after injection. In pre-treated with
diclofenac (5 mg/kg, p.o) had a significant reduction of rat paw
edema at 1 hour after the diclofenac administration. Methanolic leaf
extract at a dose of 200, 400 mg/kg, p.o exhibited the anti-
inflammatory effect to reduce the rat paw edema at 1 h after
administration and continued up to 6 hours. The inhibition of rat
paw edema of the MEQI at the dose of 200 mg/kg, p.o shows 43, 36
and 6.7% respectively while the MEQI at the dose of 400 mg/kg, p.o
decreased the rat paw edema by 43, 31, 11% respectively when
compared to the control it shows higher level of significance at the
3
rd
hour (p<0.001) as shown in Table. 2. In the present study, the
methanolic extract of Quassia indica at doses of 200,400 mg/kg
significantly decreased the rat paw edema induced by carrageenan,
suggesting methanolic extract may involve inhibition of these
inflammatory mediators release in all phases showing a good anti-
inflammatory activity Fig.2.
Table No. 1: Comparison of the DPPH free radical scavenging activity
S. No.
Concen
tration
µg/ml
% scavenging Activity
MEQI
Ascorbic acid
1
100
34.55±0.1102
***
65.013±0.013
2
200
52.86±0.0585
***
73.14±0.0100
3
300
65.65±0.0105
***
80.06±0.0200
4
400
69.86±0.0152
***
83.92±0.0152
5
500
76.29±0.0458
***
93.16±0.1528
Values are mean ± SD, n=3, p<0.001=***, indicates extremely significant increase in Scavengi ng of DPPH radical by MEQI as compared with Ascorbic acid.
Fig.1: Comparison of the DPPH free radical scavenging activity
Table No. 2 : Carragenan induced paw edema in rats
Groups
Paw volume ( ml )
% Inhibition
1h
3h
6h
1h
3h
6h
Normal
0.625 ± 0.11
0.633 ± 0.10
0.625 ± 0.11
control
1.625 ± 0.01
1.325 ± 0.02
0.875 ± 0.02
standard
0.908 ± 0.01
0.733 ± 0.01
0.625 ± 0.01
44
39
15
MEQI 200
0.933 ± 0.01
0.780 ± 0.02
***
0.691 ± 0.02
43
36
6.7
MEQI 400
0.916 ± 0.61
0.785 ± 0.02
***
0.658 ± 0.01
43
31
11
All values are Mean±SEM, n= 6, **P<0.01, *** P<0.001 represents statistical significance of treatmen t group Vs control.
Jolly John et al., J. Pharm. Res. 2014, 3(2), 11-13
Journal of Pharma Research 2014, 3(2) 11-13
Fig. 2: Effect of MEQI in carragenan induced paw edema in rats
CONCLUSION
In summary, in the present investigation it c ould be
concluded that MEQI at tested doses exhibited the pharmacological
activities; it shows the anti-inflammatory activity in acute
inflammation. The possible mechanism of action may involve
inhibition of inflammatory mediators release (histamine, serotonin,
bradykinin, prostaglandins and TNF-α in both phases. The DPPH
free radical scavenging activity results are indicating that
antioxidant principles are also having role in this leaves. Future
studies are focusing on the isolation of active constituent
responsible for the activity formulate and compare with the
commercially available preparation.
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4. Viswanad.V et.al. International Journal of Pharmaceutical
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6. OECD guideline for testing of chemicals 423.
7. Naznin et.al. Research Journal of Medicine and Medical
Sciences, 2009; 4(1): pp. 107-110.
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Conflict of interest: The authors have declared that no conflict of interest exists.
Source of support: Nil
