Anti-Inflammatory Activity of Orthosiphon Stamnineus Benth Bark Extract

 

 

Mate GS1*, Umbare RP1, Patil SM1, Dongare SS1 and Naikwadi NS2

1. A.S.P.M’s K.T. Patil College Of Pharmacy, Siddharth Nagar, Barshi Road, Osmanabad, (M.S.), India- 413501.

2. Appasaheb Birnale College of Pharmacy, Shivaji Nagar, Sangali (M.S.), India.

 

ABSTRACT

Hydro-alcoholic extract of bark of Orthosiphon stamnineus Benth (HAEOSB) was studied for its in-vivo anti-inflammatory potential using Carageenen induced rat paw edema and cotton pellet induced granuloma mehods. The result of study indicated that Hydro-alcoholic extract possess significant anti-inflammatory activity at doses 500 and 750 mg/kg.


Keywords:

 

INTRODUCTION

Plant Material:

The plant Orthosiphon stamnineus Benth (Labiatae) is commonly known as ‘Java Tea’ usually occurs in Asam, Burma, Nicobar, Islands and Deccan. The plant is traditionally used as diuretic, anti-diabetes and anti-hypertensive.1, 2

However, no systemic study on anti-inflammatory activity of the bark has been reported in the literature. In present investigation, we have screened Hydro-alcoholic extract of bark of Orthosiphon stamnineus Benth for its anti-inflammatory activity.

 

MATERIALS AND METHODS:

Bark of Orthosiphon stamnineus Benth was collected from foothill of Yercaud, Tamilnadu, India. The plant material was identified and authenticated by Dr. A. Marimuthu; Principal, Government Arts College, Attur, Tamilnadu, India.

 

Preparation of Extract:

The bark were dried under shade and then coarsely powdered with a mechanical grinder. The powder was passed through sieve No. 40 and stored in and airtight container for the extraction. The marc left after Petroleum ether extract was dried and then extracted with ethanol 95% v/v (75-780C) and distilled water mixture in proportion of 50:50 up to 72 hrs. After completion of extraction, the solvent was removed by distillation. Dark brown color residue (yield-14.85%) was obtained. The residue was then stored in a dessicator. The extract was subjected to phytochemical screening.

 

Assessment of Anti-inflammatory Activity:

Wister rats (150-180gm) of either sex and of approximately the same age, procured from listed suppliers of Venkataswara Enterprises, Bangalore, India were used for the study. They were housed in polypropylene cages and fed with standard rodent pallet diet (Hindustan Lever Limited, Bangalore) and water ad libitum. The animals are exposed to alternate cycle of l2hrs of darkness and l2hrs light. Before each test, the animals were fasted for at least 12 hrs and the experimental protocols were subjected to the scrutinization of the Institutional Animal Ethical Committee (P.Col. /14/2006) and were cleared by the same. All experiments were performed in the morning according to current guidelines for care of laboratory animals and the ethical guidelines for investigations of experimental pain in conscious animals3.  

 

 

 Table No. 01: Effect of Hydro- Alcoholic Extract of Orthosiphon stamineus Benth. on Carrageenan Induced Paw Edema in Rats

Group No.

Design of Treatment

Dose (mg/kg)

Increase in paw edema at the end of 3 hr

Percentage Inhibition

I

Control

-

0.490.023

-

II

Indomethacin

10

0.210.015**

57.87

III

HAEOSB I

500

0.320.009**

34.03

IV

HAEOSB II

750

0.280.011**

42.85

**P<0.01 Vs control, The data were statistically analyzed by Student’s t-test and all values were expressed as Mean ± SEM. The data were also analyzed by one way ANOVA followed by Dunnet’s t-test and values p<0.05 were considered significant. 

 

Table No. 02: Anti-inflammatory Activity of hydro-alcoholic Extract of Orthosiphon stamineus Benth in Cotton Pellet Induced Granuloma Model

Group No.

(n-6)

Design of Treatment

Dose (mg/kg)

Increase in paw edema at the end of 3 hr

Percentage Inhibition

I

Control

-

70.20.704

-

II

HAEOSB I

500

48.900.52*

32.41

III

HAEOSB II

750

30.420.09*

44.65

IV

Indomethacin

10

26.230.61*

55.97

 *p<0.05 Vs control, The data were statistically analyzed by Student’s t-test and all values were expressed as Mean ± SEM. The data were also analyzed by one way ANOVA followed by Dunnet’s t-test and values p<0.05 were considered significant.

 

The standard organistic  canula and syringe were used for drug administration in experimental animals.

 

Carageenen Induced Rat Paw Edema4-7:

Grouping and Treatment Protocol:

Group- I : Control group

§  Animal received carboxyl methyl cellulose (100 mg/kg P.O.)

Group- II : Standard Group

§  Rats were treated with Indomethacin (10 mg/kg I.P).

Group- III : HAEOSB – I (Test group I)

§  Rats were treated with HAEOSB (500mg/kg P.O).

Group- IV : HAEOSB – II (Test Group – II)

§  Rats were treated with HAEOSB (750mg/kg P.O).

 

In each group 6 animals were taken. Animals were kept fasted throughout the experimental period, but were provided water ad libitum.

      

After 30 min. 0.1 ml carrageenan (1%) was injected into planter region of hind paw of rats. Measurement of Paw volume (ml) was made by mercury displacement technique using Plethysmometer immediately before and 3 hr after carageenen injection.

 

 

Cotton Pellet induced Granuloma8

The animals were divided into four groups (n=6).  The animals were anaesthetized with ether, the back skin was shaved and disinfected with 70% ethanol. An incision was made in lumber region. Using a blunted forceps subcutaneous tunnels were formed and sterilized cotton pellets weighing 20+ 1 mg were implanted on either sides of the scapular region of each rat. Group-I served as control and received the vehicle.

 

The hydro-alcoholic extract at concentrations of 500 and 750 mg/kg was administered orally to Group-II and III animals for 7 days. Group-IV animals received Indomethacin at a dose of 10-mg/kg p.o. for the same period. On the 8th day, the animals were sacrificed and the pellets together with the granuloma tissues were

 

carefully removed, dried in an oven at 600C, weighed and compared with control.

 

The percentage activity of anti-inflammatory effect of hydro-alcoholic extract of Orthosiphon stamineus Benth was calculated by using following formula:

 

Percentage inhibition = C – T/C X 100

Whereas,

T- Increase in paw volume after administration of test extract

C- Increase in paw volume of control group 

 

 

Statistical Evaluation: 

The data were statistically analyzed by student’s t- test and all the values were expressed as mean  SEM. The data were also analyzed by one way ANOVA followed by Dunnet’s t- test and values p<0.05 were considered significant9, 10.

 

RESULT AND DISCUSSION:

The phytochemical and pharmacological studies on the bark of the plant Orthosiphon stamineus Benth was done revealed the presence of various phytochemical constituents like carbohydrates, glycosides, alkaloids, tannins, saponins, phenolic compounds, phytosterols and flavonoids. Hydro-alcoholic extract of Orthosiphon stamineus  Benth (HAEOSB) shows the presence of main phytoconstituent Orthosiphonin (A bitter glycoside) and Neo- orthosiphonin A – E (Flavanoid). Hence, it was selected for the pharmacological studies.

 

Administration of Carrageenan in paw edema of rats produced a short inflammatory response indicated by increase in paw volume. Oral administration of HAEOSB showed a significant (p<0.01) inhibition of carrageenan induced paw inflammation at doses 500 mg/kg (34.03% inhibition) and at 750 mg/kg (43.85% inhibition). In cotton pellet induced granuloma model the percentage inhibition (44.65) at 750 mg/kg was found significant and comparable to indomethacin at 10 mg/kg (Table-2). 

The HAEOSB showed significant anti-inflammatory activity against carrageenan induced paw edema and cotton pellet induced granuloma models in rats. The anti-inflammatory activity exhibited in this model may be attributed to the presence of flavonoids, which are found to act by reducing the release of inflammatory substance like prostaglandins there by reducing tissue exaggeration11-13.

 

The experimental results have provided the Pharmacological evidence supporting the folklore claim of the drug as an anti- inflammatory.  The results are shown in Table No. 01 and Table No.02. 

 

 

REFERENCES:

1.       Phytotherapy and Aromatherapy; Medicinal Plants, Medicinal Herbs, Essential Oils, Spices, their uses in Human and Animal Medicine; Pharmacology and Botanical Discussion; Courtesy; 360 Medicinal Plants_files/2listeplantes.htm.

2.       Kiritikar and Basu., Indian Medicinal Plants, I(2), p. 2121

3.       Ohasi K, Bohgaki T; Anti hypertensive Substances in the Leaves of  kumis Kucing (Orthosiphon aristatus) in Java Island. Yakugaku Zasshi. 120(5), 1999, p. 222-225.

4.       Bodhankar, Prasad A., Indian J. Nat. Prod., 22(1), p., 14-16.

5.       Latha, RV Manikandar., Indian J. pharm. Sci.,2006,68(4), p. 509-510.

6.       Gopalakrishna B., Sutar PS., Indian Drugs., 2006, 43(3), p. 255-257.

7.       DK Arulmozhi, SL Badhankar., Indian J. Pharmacol., 2005, 37(2), p. 96-102.

8.       Meier, R., Schuller, W. and Desulles, P.,Experientia, 1950, 6, 469.

9.       S.K.Kulkarni, Handbook of Experimental Pharmacology,Vallabh Prakashan, 3, p. 172-179.

10.     Bennet, C.A. and Franklin, N.L.,”Statistical Analysis in Chemistry and Chemical Industry”, John Wiley Sons, New York, 1967, p. 133.

11.     T.Syed Ismail, J. Ethnopharmacol., 1997, 56, p. 145. 

12.     Arrigoni-martelli, E., Spectrum, 19977, 26, p. 177.

13.     Maksimovic, Z. and Malencic, D., Bioresource Technology, 2005, 96, p. 873.

 

 

Received on 04.04.2009

Accepted on 18.05.2009     

© A&V Publication all right reserved

Research Journal of Pharmacognosy  and Phytochemistry. 1(1): July.-Aug. 2009, 18-20