Effect of leave
Extract of Prunus persica Linn on Acute Inflammation in Rats
Chiranjib
Bhattacharjee1*, Durgesh Gupta2,
Lokesh Deb3, Santhosh
Kumar C1, Subal Debnath1 and
A.S.Dutta4.
1Sri Krupa Institute of Pharmaceutical Sciences, Vil. Velkatta, Kondapak (Mdl), Dist. Medak, Siddipet. Andhra Pradesh 502277, India.
2Rajiv Gandhi
College of Pharmacy, Nautanwa, Maharajganj,
UP 273164, India.
3I.B.S.D. Takyelpat ,Imphal, Manipur, India
4R. K. Pharmacy
College, Kasipur, Suri Santhiaon, Azamgarh, UP276001, India.
ABSTRACT:
Prunus persica Linn (L) widely distributed in
Manipur, India. The leaves were used in folklore/traditional medicine to treat several inflammatory pathologies such as
greenish swelling (gland), oedema etc. Reactive oxygen species as well as
reactive free radicals such as hydroxyl (OH), nitric oxide (NO) etc. contribute
significantly to these pathologies. In this study, anti-inflammatory activity
of the aqueous extracts (AEPp) of Prunus persica L
leaves was evaluated on carrageenin induced oedema. The test sample at the dose of 200 mg/kg/p.o. were found to
cause significant (***P<0.001) inhibition of carrageenin
induced oedema.
The test samples AEPp show the protection against
inflammation less than standard diclofenac sodium 8mg/kg b.w.
These
observations established the anti-inflammatory effect of Prunus persica L leaves aqueous extract in acute inflammation.
KEYWORDS: Prunus persica, inflammation.
INTRODUCTION:
The use of plants and plant
extracts for medicinal purposes has been going on for thousands of years.
Herbarium and folk medicine both ancient and modern have been the source of
much useful therapy. Some of the plant products currently used either in their natural form or as
derivatives, were often used originally for other purposes, such as arrow
poisons, as part of religious or other rituals and even as cosmetics. With this
background in present study we thought of finding a remedy available at a
hands stretch for the treatment and management of inflammatory response. In this connection we had undertaken field surveys and
contact programmes with native practitioners of
Manipur to explore the possibilities of using locally available herbs for the
purpose. In two of our field surveys we found a plant Prunus persica L and native practitioners were claimed that it is highly
useful in treating inflammatory disorders. In our present study used animal
model to evaluate Prunus persica L. The Carageenan
induced inflammation is a useful model to detect oral action of
anti-inflammatory agents. The development of carrageenin induced edema is believed to be biphasic of
which the first phase is mediated by release of histamine, serotonin, and kinins in the first hour after injection of carrageenan and the second phase is related to release of
prostaglandin like substances in 2-3 hours 1.
In this project
we have chosen a plant Prunus persica
L (Family : Rosaceae) which is very popular as a
medicinal agent as revealed in a ethno pharmacological survey conducted in
different district of Manipur by team of scientists, IBSD, Imphal.
Traditional
practitioners of the region used leaves of this plant as antihypertensive
agent; its also giving edible fruits2.
MATERIALS AND METHODS:
Plant Material:
Prunus persica L leaves were
collected from the garden of IBSD, Imphal. The plant
was identified and authenticated by Dr. Biseswhori Thongam, Scientist C (Plant Taxonomy), IBSD, Takyelpat, Imphal, Manipur where
a voucher specimen were deposited for reference to Plant Taxonomy and
conservation Lab, IBSD, Takyelpat, Imphal (IBSD/M/1019). The leaves were shade dried at
room temperature. The powder obtained
was subjected to soxhlet extraction with the water as
solvent. Aqueous extract divided in two equal volumes. One portion concentrated
in vacuum evaporator and dried in desiccators and other portion was mixed with
equal quantity of petroleum ether and vigorously shack in separating funnel to
separate aqueous and petroleum ether portions. The aqueous extract was used for
anti-inflammatory studies3. The products were concentrated under
reduced pressure and stored in refrigerator 8 ± 2° C.
Animals:
Albino rats (Wister) weighing
150-200g and albino mice weighing 20-25g of either sex were used in this study.
They were procured from Regional Institute of Medical Sciences (RIMS), Imphal. The animals
were acclimatized for one week under laboratory conditions. They were housed in
polypropylene cages and maintained at 27°C ± 2°C under 12 hours dark / light. They were fed with soya bean chock, Gram and
water ad libitum was provided. The litter in the cages was renewed daily to
ensure hygienic condition and maximum comfort for animals. Ethical clearance for handling the animals
was obtained from the Institutional Animals Ethical Committee (IAEC), IBSD, Imphal prior to the beginning of the project work.
Determination of acute toxicity (LD50):
The acute toxicity for aqueous extracts (AEPp) of Prunus persica L leaves was
determined in albino mice, maintained under standard conditions. The animals
were fasted overnight prior to the experiment. Fixed dose (OCED Guideline no.
420) method of CPCSEA was adopted for toxicity studies (Mrs Prema Veeraraghavan, 2003). The tested
extracts were administrated orally. No mortality was observed at 2000mg/kg in
the all cases4.
Evaluation of Anti-inflammatory Activity:
Carrageenan - induced paw edema:
The
animals were divided into seven groups of 5 animals each. Inflammation was
induced by injecting 0.1ml of 1% w/v carrageenan
sodium salt subcutaneously in the sub-plantar region of the rat right hind paw
in each groups.
Group-I - Animals
(Control) were administered 1ml distill water p.o.,/animal
Group-II -Animals
were administered with diclofenac sodium 8mg/kg b.w.
Group-III -
Animals were administered with aqueous extracts 200mg/kg/ p.o.,
1
hour after oral administration of reference and test drugs, carrageenan
was injected. The hind paw volume was measured plethysmometrically
before and after the carrageenan injection, at hourly
intervals for 6 hrs1.
Where,
VT = mean paw volume of test group.
VC
= mean paw volume of control group.
RESULTS AND DISCUSSION:
In
acute toxicity study of aqueous extracts (AEPp) and
of Prunus persica L leaves dose not shown
mortality at the dose of 2000 mg/kg. Therefore 2000 mg/kg dose was consider as
ALD50 cut off the dose (safe dose) so 1/10 of that dose was selected
(200 mg/kg) for in vivo experiments. The aqueous extracts (AEPc)
of Prunus persica L leaves cause significant (***P<0.001) inhibition of carrageenin
induced edema.
The current study establishes the anti-inflammatory
activity of the aqueous extract of Prunus persica L leaves, at 200 mg/kg b.w. dose,
employed for screening of inflammatory process. In the best concerning carrageenan induced, the extract was found to possess
significant (*p<0.001) anti-inflammatory effects but less potent then diclofenac sodium (Table
1). Since there are reports that at sites of
inflammation, increased free radical activity is associated with the
activation of the neutrophil NADPH oxidase and/or the uncoupling of a variety of redox systems, including endothelial cell xanthine dehydrogenase. Although
free radicals, thus produced, have the capacity to mediate tissue destruction,
either alone or in concert with proteases, it was argued that
disturbances in the second messenger and regulatory activities of
free radicals may also contribute significantly to the inflammatory process5.The
aqueous extracts (AEPp) of Prunus persica L leaves caused significant
anti-inflammatory effect in the acute inflammatory model of in rats. Reactive
Oxygen species (ROS) generated endogenously or exogenously are associated with
the pathogenesis of various diseases such as atherosclerosis, diabetes, cancer,
arthritis and aging process. Inflammation is a complex process and ROS play an
important role in the pathogenesis of inflammatory diseases. Thus antioxidants
which can scavenge ROS are expected to improve these disorders6.The
aqueous extracts (AEPp) of Prunus persica L leaves also process antioxidant activity in our laboratory. However
results are not included here.
Table 1: Effect of Prunus persica
L on carrageenin
induced paw edema in rats
|
Treatment
|
Mean paw volume (ml) ± SEM
|
|
0 hrs
|
1 hrs
|
2 hrs
|
3 hrs
|
4 hrs
|
6hrs
|
|
Normal
control
(1ml dist.
water p.o.)
|
0.24 ±
0.02449
|
0.4 ±
0.02739
|
0.54 ±
0.02915
|
0.62 ±
0.04665
|
0.54 ±
0.04301
|
0.52 ±
0.03391
|
|
Standard Diclofenace Sodium (8 mg/ kg p.o.
)
|
0.22 ±
0.02000
|
0.37 ±
0.02550
|
0.42 ±
0.02550
|
0.41 ±
0.04000
|
0.31 ±
0.03317
|
0.28 ±
0.02550
|
|
Aqueous
Extract (200 mg/kg p.o.) Prunus
persica
|
0.26 ±
0.04000
|
0.34 ±
0.04944
|
0.39 ±
0.05988
|
0.43 ±
0.06317
|
0.39 ±
0.05988
|
0.38 ±
0.05409
|
CONCLUSION:
At sites of
inflammation, increased free radical activity is associated with the activation
of the neutrophil NADPH oxidase
and/or the uncoupling of a variety of redox systems,
including endothelial cell xanthine dehydrogenase. Our
present study revealed that Prunus persica L able to cause protection against above mentioned pathologies of
inflammatory disorder in experimental animals that also claimed by traditional
practitioners. Prunus persica L leaves may safe anti-inflammatory for next generation. However, further
work on phyto-chemicals present in the plant and its
details pharmacological properties is carrying out in the pharmacology
laboratory, IBSD, Imphal.
REFERENCES:
1.
Deb Lokesh, Jain Avijeet, Porwal Piyush, Talera Deepti, Dutta Amitsankar.
(2007) Protective effect of Eucalyptus globulus Labill on acute and chronic inflammation in rats. Indian drug,
44(10): 774 -777.
2.
M. Samarjit Singh, N. Rajendro
Singh (2008) Plants used as traditional medicine in Manipur. Asian Agro-History
12(2); 153-156.
3.
Plummer SM,
Holloway KA, Manson MM, et al. Inhibition of cyclo-oxygenase
2 expression in colon cells by the chemopreventive
agent curcumin involves inhibition of NF-kappaB activation via the NIK/IKK signalling
complex. Oncogene. 1999; 18: 6013-6020.
4.
OECD Organization for Economic
Co operation and Development (1997) Test No. 420: Acute oral toxicity fixed
dose procedure OECD Guidelines for the testing of chemicals. 1(4): 1-14.
5.
Amann R., Schuligoi R., Lanz
I., Donnerer J., (1995) Histamine induced edema in
the rat paw effect of capsaicin denervation and cgrp receptor agonist, Europian
Journal of Pharmacology. 279; 227-231.
6.
Porchexhian E, Ansari SH. Hepatoprotective
activity of Abutilon indicum on experimental damage
in rats. Phytomedicine 2005; 12: 62-64.