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,
14 November 1990
|
