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Research
Findings
Anti-Inflammatory
and Anti-Pyretic Action of Juniperus Communis. Linn. Leaf Extract
in Rats
Juniperus
communis, Linn is a plant widely available in the northern part
of India mainly at the altitude of more than 1350 m. The different
parts of the plant have been claimed to possess medicinal properties
in traditional medical system. Traditionally the plant is used by
Tibetian in gastric and urinary troubles1. A few reports concerning
the activity of J. communis, L. are available2-4; so we have studied the pharmacological
action of leaf extract of J. communis L. and we wish to report
here the anti-inflammatory and anti-pyretic action of methanol fraction
of the leaf extract.
Leaves
of the plant J. communis L. was dried at 60°C in an air oven:
the dried leaves were powdered in a grinder; this was then extracted
in a soxhlet extractor with methanol. Finally, the methanolic extract
was dried in vacuo. The sticky blakish-green material obtained
after drying was partially purified in a column (silica gel G 60
type) using methanol: chloroform solvent system (1:1). After column
chromatography, the plant extract was completely dried to constant
weight in vacuo. This was kept at 4°C in a desicator for
further use and just before pharmacological testing, was dissolved
in hot distilled water (at 60°C).
The
anti-inflammatory studies were conducted using male rats of Charles-Forster
strain (120-180 g). For the chronic experiments, the animals had
free access to food and water throughout the period. For the acute
experiments, the rats were fasted for 18 hr., but water was allowed
ad lib.
Carrageenin
(0.1 ml of 1% solution) was injected into planter aponeurosis of
the right hind paw of rats. The animals received the drugs or control
vehicle i.p. 30 minutes before the injection of carrageenin, The
paw volume was measured before and 3 hour after carrageenin administration
by volume displacement method5-7.
Test
doses of plant extract (50 mg/kg and 100 mg/kg) were given orally
against carrageenin-induced paw oedema. For differentation of counter-irritant
activity and 'true' anti-inflammatory activity the extract and carrageenin
were mixed (mixture I, 5 mg extract and 0.1 ml 1% carrageenin solution;
mixture II, 10 mg of extract and 0.1 ml of 1% carrageenin solution),
the mixtures were administered to the hind paw and the paw volume
was measured as described above.
The
method of Meier, Schaler and De Saulles9 as described by Finney and
Somers10 was adopted with slight modifications.
Sterile cotton pellets (weighing, 10 mg each) were implanted subcutaneously
along the flanks or axillae of rats. Drug extract (50 mg/kg &
100 mg/kg) or phenyl butazone (100 mg/kg) or control vehicle were
administered i.p. for 7 days from the day of cotton pellet implantation.
Granuloma was measured by weighing the implanted cotton pellets
after their removal, on the eighth day. The pellet were freed from
extraneous tissue and dried at 60°c to constant weight.
Male rats were
injected s.c. with 15% suspension of brewers yeast (1 ml/100 gm). After 15 hours the body
temperature of each animal was measured rectally with a electronic thermometer (model No.
DCT/1002). This served as the initial pyretic temperature, when drug extract (50 mg/kg and
100 mg/kg) or Aspirin (100 mg/kg) or control vehicle (0.2 ml) was administered i.p. Rectal
temperature was recorded at hourly intervals for the subsequent 4 hours11.
In all
the experiments the effect of the plant extract was compared with
standard (either phenylbutazon or aspirin 100 mg/kg) anti-inflammatory
antipyretic drugs.
The
data were analysed statistically using student's t-test.
The plant extract in
the doses employed (50 mg,100 mg/kg body wt) significantly (P < 0.001) inhibited the
carrageenin induced oedema though less effectively than phenyl butazone (100 mg/kg).
Simultaneous injection with carrageenin into the paw of the rats did not exhibit and
counter-irritant activity of plant extract.
The plant extract
was found to reduce the weight of cotton pellet induced granuloma in rat significantly (P
< 0.001).
The
effect of the plant extract and acetylsalicylic acid, (aspirin)
on yeast induced pyrexia in rats is encouraging. It is evident that
the plant extract possessed a significant antipyretic activity (P
< 0.001 at 1, 2, 3 and 4 h).
From
the battery of pharmacological tests we can conclude that our test
plant extract of J. communis, L. positively possesses anti-inflammatory
and anti-pyretic activity. In the tests concerning carrageenin-induced
oedema, the extract was found to possess significant anti-inflammatory
activity although less potent than the standard drug, phenylbutazone.
It is established that anti-inflammatory substances which exert
the!r effects by virtue of their irritant properties can be distinguished
from true anti-inflammatory agents by administering them locally
in the carrageenin test. The effects of the plant extract are not
due to counter-irritant activity, since a mixture of plant extract
and carrageenin produced a reduction in paw oedema. While it is
difficult to give an adequate description of the inflammatory phenomenon
in terms of underlying cellular events in the injured tissue, there
are certain features of the process that are generally agreed to
be characteristic.
These
include penetration of the microvasculature, leakage of the elements
of the blood in the interstitial spaces and migration of leukocytes
in the inflammed tissue. On a microscopic level this is usually
accompanied by the familiar clinical signs of erythema, oedema,
tenderness (hyperalgesia) and pain. The plant extract may interfere
with anyone or simultaneously all aforesaid processes of inflammation
and act as an anti-inflammatory substance. The repairing phase of
inflammation is initiated as a proliferation of fibroblasts and
a multiplication of small blood vessels. Proliferating cells penetrate
the exudate, producing the highly vascularized reddened mass known
as "granulation tissue"12. Significant reduction of
cotton pellet-induced granuloma in rats, by the plant extract would
suggest an activity in the proliferative phase of inflammatory process.
There is evidence
that mycobial endotoxin (lipo-polysaccharides from the cell wall) act by stimulating the
bio-synthesis and release by neutraphils and other cells of an endogenous pyrogen, a
protein with molecular weight in the range of 10,000 to 20,000. The current view is that
the endogenous pyrogen passes from general circulation in the central nervous system,
where it acts upto discrete thalamic area. There is evidence that the resultant elevation
of body temperature is mediated by the release of prostagrandins. And aspirin supress the
effects of endogenous pyrogen by inhibiting synthesis of these substances13. It may be possible that plant extract
also interfered with prostaglandins, synthesis and reduced yeast-induced pyrexia.
Authors
wish to thank University Grants Commission (UGC) New Delhi for financial
assistance rendered. They are also grateful to Dr. R. B. Ghosh,
Regional Botanist of Botanical Survey of India, Sibpur for identification
of plant.
References
1.
In "Medicinal Plant of India". Vol. -2, (ICMR)
1987, P. 106-107.
2. Devi, G. and Sisodia, C. S Indian J. Anim. Sci., 1969,
39, 345.
3. Srivastava, S. C. and Sisodia, C, S., Indian Vet. J.,
1969, 46, 826.
4. Aswal, B. S., et. al, Indian J. Exp. Biol. 1984, 22,487.
5. Winter, C. A., Risley, E. A. and Nuss. G, W., Proc. Soc. Exp.
BioI. Med., 1362 111 544-547.
6. Pillai, N. R, and Santhakumari, G., Planta Medica, 1981.
43, 59-63.
7. Bhatt, K. R.. Mehta, R. K. and Shrivastava, P.N., Indian J.
Physiol. Pharmacol., 1977, 21, 399. 400.
8. Shanahan, R. W ., Arch. Int. Pharmacodyn., 1968, 175,
186-192.
9. Meier, R, W ., Schular and De Saulles, P., Experientia,
19,50, 6, 469-471.
10. Finney, R.S.H. and Somers, G. R., J. Pharm. Pharmac.,
1958, 10, 613.620.
11. Gujral, M. L., Kohli, R. p. and Saxena, P.N., Indian J. Med,
Res., 1955, 43, 89-94.
12. Swingle, K.F., In " Anti.inflammatory Agents : Chemistry
and Pharmacology", (Scherrer, R. A. and Whitehouse, M.
W. eds.) 1974, P. 33.122, Vol. 2.
13. Flower, R.J., Moncada, S. and Vane, J, R., In "Drug
therapy of inflammation: Goodman and Gilman's the pharmacological
basis of therapeutics" (Macmillan publishing company, NY)
r. 674.715, 7th ed.
* Dept. of Pharmaceutical
T. Chatterjee*1
Technology, Jadavpur University C.
Ghosh**
Calcutta.700 032,
P. Raychaudhuri * *
** Dey's Medical Stores (Mfa) Ltd.,
62 Bondel Road, Calcutta-700 019,
1 For Reprints
14 November 1990
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