Contact Us | FeedbackSitemap
Search by Ailments
 
   
   

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 60C 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 4C in a desicator for further use and just before pharmacological testing, was dissolved in hot distilled water (at 60C).

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 60c 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