INTRODUCTION:

Plant:

Grewia tiliaefolia (Tiliaceae) aerial parts were collected from botanical garden and authenticated by Dr. V. Chelladurai, Research Officer, Botany, CCRAS, Govt. of India. The plant materials were deposited in our college herbarium for future reference.

The parts of the plant Used in traditional medicine are root, bark and fruit. They are available throughout India in deciduous and semigreen forest. The bark is used to treat cough, skin diseases, wound, ulcer and general debility. Traditionally it is used to treat inflammation⁴. A bibliographical survey showed that there is no report on in vitro anti arthritic activity of the methanolic extract of the selected plant.

Reported Activity: Anti inflammatory³, wound Healing and anti bacterial.

EXPERIMENT:

Extraction: Grewia tiliaefolia (Aerial parts) was dried and powdered. The extraction procedure was carried out by cold maceration process based on increasing polarity. Solvent was concentrated under reduced pressure to get crude extract which was stored under reduced pressure to get crude extract which is stored in desiccators for future use.

MATERIALS AND METHODS:⁵

Procedure:

1. Inhibition of protein denaturation

1. Test solution (0.5ml) consist of 0.45 ml of bovine serum albumin (5% w/v aqueous solution) and 0.05 ml of test solution of various concentrations

2. Test control solution (0.5ml) consist of 0.45 ml bovine serum albumin (5%w/v aqueous solution) and 0.05 ml of distilled water

3. Product control solution (0.5ml) consist of 0.45ml of distilled water and 0.05ml of test solution (250µg/ml)

4. Standard solution (0.5ml) consists of 0.45ml of bovine serum albumin (5%w/v aqueous solution) and 0.05ml Diclofenac sodium solution (250µg/ml).

The pH of all solutions is adjusted to 6.3 by using a small amount of 1N hydrochloric acid. All the samples are incubated at 37ºC for 20 minutes and heated at 57ºC for 3 minutes. After cooling add 2.5ml of phosphate buffer to above solution. The absorbance was measured using UV visible spectrophotometer at 416nm. The percentage protein denaturation can be calculated by

Percentage inhibition =

100 - (O D of test solution-O D of product control) Ï100

O D of test control

The control represents 100% protein denaturation .The results were compared with Diclofenac sodium (250µg/ml).

2. Effect of membrane stabilization

The reaction mixtures (4.5ml) consist of 2 ml hypotonic saline (0.25% NaCl), 1ml of 0.15m phosphate buffer pH 7.4 and 1 ml of test solution in normal saline. 0.5ml of 10% RBC in normal saline was added. For control, 1 ml of isotonic saline was added instead of test solution. RBC was lacked in product control. Mixture was incubated at 56ºC for 30 minutes. The tubes were cooled under running tap water for 20 min. The mixture was centrifuged and the absorbance of the supernatant liquid was read at 560nm. The percentage membrane stability was calculated as follows.

Percentage membrane stabilisation =

100 - (O D of test solution-O D of product control) Ï100

O D of test control

The control represents 100% lysis .The results are compared with Acetyl salicylic acid (250µg/ml).

RESULTS AND DISCUSSION:

Anti arthritic effect of Grewia tiliaefolia (Aerial parts) was studied by testing various in vitro studies. The effect of the selected plant on inhibition of protein denaturation and membrane stabilization was 85.46% and 86.88% respectively for the methanolic extract of the selected medicinal plant. The results were compared with the standard drugs Diclofenac sodium and acetylsalicylic acid at the concentration of 250µg/ml. The results are tabulated in Table 1and 2.

Most investigators have reported that denaturation of proteins is one of the cause of rheumatoid arthritis. Production of auto antigen may be due to denaturation of proteins. Protective effect on heat and hypotonic saline induced erythrocyte lysis is known to be a very good index for anti arthritic activity.

Table 1: In Vitro anti arthritic activity of Grewia tiliaefolia –Protein denaturation method

S. NO

CONTENTS

CONCENTRATION (µg/ml)

% PROTEIN INHIBITION

STANDARD

(Diclofenac sodium)

250µg/ml

88.12 %

TEST SAMPLE

a. Methanolic extract

b. Petroleum ether extract

250µg/ml

85.46 %

65.23 %

Table 2: In Vitro anti arthritic activity of Grewia tiliaefolia –Membrane stabilization method

S. NO

CONTENTS

CONCENTRATION (µg/ml)

% MEMBRANE STABILISATION

STANDARD

(Acetyl Salicylic Acid)

250µg/ml

87.65 %

TEST SAMPLE

a. Methanolic extract

b. Petroleum ether extract

250µg/ml

86.88 %

66.34 %

CONCLUSION:

The in vitro studies which were carried out by above mentioned methods in methanolic extract possess anti arthritic activity and it was comparable to the standard drug. Future research can be extended to study the in vivo anti arthritic activity in animal models

REFERENCES:

1. Vaidhyranthnam, Ps Warrier, Indian medicinal plants, a compendium of 500 species, Vol.3,Arya Vaidya Sala, Kottakal, Orient longman,pp-104,1996.

2. Badami et al, Pharmacognostical evaluation of Grewia tiliaefolia bark, Indian journal of natural products,Vol.18(2) pp-6-11,2002.

3. Sakat S.S et al, Anti inflammatory potential of aqueous extract of Grewia tiliaefolia leaves, Indian drugs, Vol 47(1)pp-36-43,Jan 2010.

4. Ethnomedical information from Kattunayakas tribes of Mudumalai Wildlife Sanctuary, Nilgris, Tamilnadu, Indian journal of Traditional knowledge, Vol (6)pp-574-578,oct 2007

5. Gopinathan et al Investigation of in vitro anti arthritic ,anti inflammatory and anti oxidant activity of Tectona garndis, Indian drugs, Vol 47(6)June 2006.

Received on 26.07.2011 Modified on 04.08.2011

Research J. Pharm. and Tech. 4(12): Dec. 2011; Page 1833-1834