Antipyretic Activity of Azima tetracantha in Experimental Animals

 

T. Nargis Begum1, A. Vijaya Anand2 and R. Senthil2

 

1Post Graduate Department of Biotechnology, Jamal Mohamed College, Tiruchirappalli-620 020, Tamil Nadu, India.

2Department of Biochemistry, M.I.E.T. Arts and Science College, Tiruchirappalli-620 007, Tamil Nadu, India.

 

 

ABSTRACT:

In the present study, the ethanolic leaf extract of Azima tetracantha Lam (A. tetracantha) was investigated for antipyretic activity in rats using Brewer’s yeast induced Pyrexia. The leaves of A. tetracantha were collected from Ponnamaravathi and cut into small pieces and shade dried. The dried powdered leaves (100 gm) were extracted in a Soxhlet apparatus by using 95% ethanol. Albino rats weighing (200-250g) were taken for the experiment divided into four groups of six animals each. Group 1 received 3% aqueous suspension of gum Acacia (1 ml/200g) as vehicle orally, group 2 and group 3 received ethanolic leaf extract of A. tetracantha 100 and 200 mg/kg with 3% aqueous suspension of gum Acacia orally and the group 4 served as standard received paracetamol 25 mg/kg with 3% aqueous suspension of gum Acacia orally. The subcutaneous injection of yeast suspension markedly elevated the rectal temperature after 18h of administration. Treatment with A. tetracantha extract at a dose of 100, 200 mg/kg decreased the rectal temperature of the rats in dose dependent manner.  This effect was maximal at dose of 200 mg/kg and it caused significant lowering of body temperature (P< 0.01) up to 4 hour after its administration.  The antipyretic effect started as early as 1h and the effect was maintained for 4h, after its administration. Both the standard drug paracetamol 25 mg/kg and tested drug A. tetracantha extract were significantly reduced the yeast elevated rectal temperature, at 2nd, 3rd and 4th hour compared to control group.

 

KEYWORDS: Azima tetracantha, antipyretic, Brewer’s yeast, paracetamol,

 

INTRODUCTION:

Pyrexia or fever is caused as a secondary impact of infection, malignancy or other diseased states. It is the body’s natural defense to create an environment where infectious agent or damaged tissue cannot survive1. Normally the infected or damaged tissue initiates the enhanced formation of pro-inflammatory mediator’s (Cytokines like interleukin 1β, α, β and TNF- α), which increase the synthesis of prostaglandin E2 (PG E2) near peptic hypothalamus area and thereby triggering the hypothalamus to elevate the body temperature2. As the temperature regulatory system is governed by a nervous feedback mechanism, so when body temperature becomes very high, it dilate the blood vessels and increasing sweating to reduce the temperature; but when the body temperature become very low hypothalamus protect the internal temperature by vasoconstriction. High fever often increases faster disease progression by increasing tissue catabolism, dehydration and existing complaints, as found in HIV3. Drugs having anti-inflammatory activity generally possess antipyretic activity (e.g) non-steroidal anti-inflammatory drugs (NSAIDs).

 

 

It has been suggested that prostaglandin (PGE) mediates pyrogen fever; the ability of NSAIDs, to inhibit prostaglandin synthesis could help to explain their antipyretic activity.

 

Search for safe herbal remedies with potent antipyretic activity received momentum recently as the available antipyretics, such as paracetamol, aspirin, nimusulide etc, which have toxic effect to the various organs of the body4. The subacute toxicity results revealed that Azima tetracantha (A. tetracantha) might be considered as a broad non-toxic one. The antipyretic activity exhibited that the ethanol extract of leaves possess a significant antipyretic effect in maintaining normal body temperature and reduced the elevated rectal temperature in rats and their effects are comparable to that of standard antipyretic drug paracetamol. Such reduction of rectal temperature of the tested animals appears to be due to the presence of a single bioactive substance or a mixture of compounds in them. Therefore, the present study aimed to evaluate the analgesic effect of ethanolic leaf extract of A. tetracantha.

 

ATERIALS AND METHODS:

Collection and Extraction:

Fresh leaves of A. tetracantha were collected in Ponnamaravathi (Pudukkottai District) during the month of November-December. The drug was authenticated by botanist at the Rapinat Herbarium and Centre for Molecular Systemics, St. Joseph College Tiruchirappalli, Tamil Nadu, India.  Plant material was dried under shade at room temperature, pulverized by a mechanical grinder, sieved through 40 meshes. The powdered material (100 g) was extracted with 95% ethanol by hot continuous Percolation method in a Soxhlet apparatus. The extract was then concentrated and dried under reduced pressure. The ethanol free semi solid mass obtained (13.65g) and suspended in 5% gum Acacia for pharmacological studies. This study was carried out in the animal house of Periyar College of Pharmaceutical Sciences for Girls, Tiruchirappalli (Regd. No. 265 / CPCSEA). Toxicity study was carried out as per the organisation for Economic Co-operation and Development (OECD) guidelines. The LD50 of the A. tetracantha ethanolic leaf extract as per OECD guidelines falls under class 4, values with no signs of acute toxicity LD50> 2000 mg/kg). Hence the dosage was fixed in 100 and 200 mg/kg b.w.

 

Animals:

Swiss albino mice of both sexes weighing between (18-25 g) were used for the experiment.  The animals were kept in clean and dry plastic cages, with 12h: 12h light dark cycle at 25±2°C temperature and 45-55% relative humidity.  The animals were fed with standard pellet diet and water was given ad libitum.

 

Antipyretic activity: 5

Brewer’s yeast induced Pyrexia in Rats:

Antipyretic activity on albino rats were studied with fever induced by 20% Brewer’s yeast.  Albino rats (200-250g) were fed uniformly till 24 hours, and food was withdrawn before giving drugs. After measuring rectal temperature of the rats by introducing 1.5 cm of digital thermometer in rectum, pyrexia was induced by injecting subcutaneously, 20% suspension of dried yeast in 2% gum Acacia in normal saline at a dose of 20 ml/kg of body weight. After 18 hour of yeast injection, rats which showed a rise in temperature of at least 1°C were taken for the study. Animals in the various groups were treated as follows:

 

Group 1: 3% aqueous suspension of gum Acacia (1 ml/200g) as vehicle, orally.

Group 2: Ethanolic leaf extract of A. tetracantha 100 mg/kg (1 ml/200g) with 3% aqueous suspension of gum Acacia, orally.

Group 3: Ethanolic leaf extract of A. tetracantha 200 mg/kg (1 ml/200g) with 3% aqueous suspension of gum Acacia, orally.

Group 4: Paracetamol 25 mg/kg (1 ml/200g) with 3% aqueous suspension of gum Acacia, orally

Rectal temperature was recorded every hour for four hours after administration of drugs.

 

RESULT:

Effect of ethanolic leaf extract of A. tetracantha on rectal temperature in rats is presented in Table 1. The subcutaneous injection of yeast suspension markedly elevated the rectal temperature after 18h of administration. Treatment with A. tetracantha extract at a dose of 100, 200 mg/kg decreased the rectal temperature of the rats in dose dependent manner. It was found that the extract at a dose of 100 mg/kg caused significant lowering of body temperature at 4 hour following its administration (36.91 ± 1.15). This effect was maximal at dose of 200-mg/ kg and it caused significant lowering of body temperature (P< 0.01) up to 4 hour after its administration (36.16 ± 0.15). The antipyretic effect started as early as 1h and the effect was maintained for 4h, after its administration. Both the standard drug paracetamol 25 mg/kg and tested drug A. tetracantha extract were significantly reduced the yeast-elevated rectal temperature, at 2nd, 3rd and 4th hour compared to control group.

 

DISCUSSION:

A. tetracantha leaf extract possess alkaloids, flavonoids, tannins, β-sitosterol, terpenes, protein, coumarin, glycosides, protein and starch. The β-sitosterol is a plasminogen activator and promotes the formation of essential polyunsaturated fatty acids from linoleic acid, but linoleic acid is required for prostaglandin and leukotrienes synthesis6 and thus beta sistosterol reduces prostaglandin and leukotrienes synthesis. β-sitosterol possesses potent anti-inflammatory and antipyretic activity7 by reducing the secretion of proinflammatory cytokines and alpha-TNF7,8.


Table 1 Effect of ethanolic leaf extract of A. tetracantha on Brewer’s yeast induced   Pyrexia in Wistar albino rats

Treatment

 

Dose

Normal Temperature  (°C)

Rectal Temperature (°C) 18 hrs after yeast induced pyrexia

Rectal temperature (°C) after treatment with extract

1h

2h

3h

4h

Control

-

35.23 ± 0.13

38.08 ± 0.18

38.09 ± 0.16

38.05 ± 0.17

37.98 ±

0.17

37.99±

0.27

A. tetracantha ethanolic leaf extract

100 mg/kg

35.49 ± 1.03

37.90 ± 2.10

37.7 ± 1.09

37.03 ± 0.95

36.94 ±

0.04

36.91±

1.15

A. tetracantha ethanolic leaf extract

200 mg/kg

35.29 ± 0.23

38.02 ± 0.19

38.04 ± 0.13

37.45 ± 0.10

36.53* ±

0.20

36.16* ±

0.15

Paracetamol

25 mg/kg

35.13 ± 0.25

38.01 ± 0.19

37.9 ± 0.08

37.26 ± 0.07

36.65* ±

0.15

35.88** ±

0.08

Values  are expressed as Mean ±S.E, n =6 by students ″ t ″ test

*P< 0.01 Vs control, ** P< 0.001 Vs control

 

These phytosterols can enhance adaptive immunity through the stimulation of innate immune system termed as the “adaptogen” which promotes overall health without side effects9.

 

In general, non-steroidal anti-inflammatory drugs produce their antipyretic action through inhibition of prostaglandin synthetase within the hypothalamus10. The ethanolic, butanolic and petroleum ether extracts of dried leaves of Pergularia extensa showed significant antipyretic activity in rats is due to the presence of the phytoconstituents flavanoids, steroids and saponins11.  Presences of flavonoids were reported in Dalbergia species and flavonoids are known to inhibit prostaglandin synthetase12. Therefore it appears that antipyretic action of Dalbergia species may be related to the inhibition of prostaglandin synthesis in hypothalamus13.  The antipyretic properties of Acacia catechu may be ascribed to the presence of flavonoids14. As some flavonoids are predominant inhibitors of cyclooxygenase or lipooxygenase15,16.

 

Chloroform extract of the Solanum nigrum leaves exhibited antipyretic activity when assessed against Brewer’s yeast induced pyrexia test is due to the presence of phyto constituents like steroidal glycosides and steroidal oligoglycosides17. Myrica salicifolia root extract was found to have analgesic and antipyretic activity in mice. The phytoconstituents responsible for these activity is a variety of flavonoids among which myricitrin is generally considered18.

 

The preliminary phytochemical screening of the petroleum ether and chloroform fraction of root of Laportea crenulata showed the presence of steroids, tannins and flavonoids19. The antipyretic activity of Laportea crenulata is due to the presence of steroids20.  In many studies, flavonoids have been reported to exhibit antipyretic effect21,22. Aqueous extract of Untica macrorrhiza a related species of Laportea crenulata suppressed yeast induced fever in rats 23. The dry residue of fresh juice produced significant antipyretic effect in a dose dependent manner. The phytochemical analysis of the dry residue showed the presence of flavonoids, alkaloids, tannins and steroids. The antipyretic activity observed can be attributed to the presence of flavonoids24.

 

Isoflavones have antipyretic effects25. Kerr and Collaborators 198126, had isolated 12 different flavonoids from Neurolaena lobata and suggested that these could be some of the components responsible for Neurolaena lobata antipyretic effect. In a similar, but more detailed study was shown that Pueraria lobatas isoflavones were responsible for the antipyretic activity27.

 

CONCLUSION:

The results of the present study suggest that the ethanolic leaf extract of A. tetracantha in doses of 100 and 200 mg/kg, significantly reduce the temperature of pyretic rats as revealed from the observation that the average percentage of antipyretic activity increased with the concentration of the extracts (200mg/Kg) compared with the control. It is also presumed that the presence of flavonoids may be contributing to antipyretic activities of ethanolic leaf extract of A. tetracantha in addition to the analgesic effect, as in the case of many of the established antipyretics.

 

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Received on 16.09.2010

Accepted on 30.09.2010        

© A&V Publication all right reserved

Research Journal of Pharmacognosy  and Phytochemistry. 2(6): Nov. - Dec. 2010, 451-454