INTRODUCTION:

Rheumatoid arthritis is an autoimmune disease in which there is joint inflammation, synovial proliferation and destruction of articular cartilage¹ .Inflammation is a bodily response to injury, infection or destruction characterized by heat, redness, pain, swelling and disturbed physiological functions. It is a common disease having peak incidence in 3rd to 4th decades of life with 3 - 5 times higher preponderance in female². The commonly used non - steroidal anti - inflammatory drugs (NSAIDs), leads to formation of gastric ulcers. Herbal drugs constitute a major part in all the traditional system of medicine^3,4.

Rice is one of the most important stable foods in Asia. India has one of the oldest, richest and most diverse cultural traditions associated with the use of medicinal rice. The Indian Materia Medica shows the existence of many medicinal rice varieties in India⁵. Njavara belongs to the family Oryzae. It is grown exclusively in Kerala, South India, since ancient times, and is used mainly for ayurvedic treatments. Njavara is an upland crop and cultivated in a water-stressed environment. It is belived that Njavara is used to treat arthritis, cervical spondylitis, muscle wasting, skin diseases, certain neurological problems circulatory, respiratory and digestive systems. Njavara kizhi is a specialized Ayurvedic therapy for treatment of paralysis, arthritis and neurological problems⁶. Based on the previous evidence the in vitro Anti Arthritic activity was analyzed by protein denaturation method.

MATERIALS AND METHODS:

Collection of Sample

Plant material Njavara was obtained from Musiri. Rice from the sample was obtained by milling rice grain in a local grinding mill, followed by sieving to separate grain from bran.

Preparation of extract

Rice powder (5 g) was extracted thrice with 45 ml Ethanol for 3 h in an electrical shaker at 40°C. The extracts were filtered through Whatmann No.1 filter paper and evaporated. The extracted sample was used for the following analysis.

IN-VITROANTI- ARTHRITIC ACTIVITY:

Anti-denaturation study is performed by using bovine serum albumin (BSA) . When BSA is heated it undergoes denaturation and express antigens associated with type - III hypersensitivity reaction and that is related to disease such as serum sickness, golmeralonephritis, rheumatoid arthritis and system lupus eruthematosus⁷.

INHIBITION OF PROTEIN DENATURATION METHOD

Materials and method

Preparation of reagents

·5% Bovine serum albumin (BSA)- 5g of BSA was dissolved in 100ml of water.

·Phosphate buffer saline pH 6.3 –8g of sodium chloride (NaCl), 0.2 g of potassium chloride (KCl), 1.44g of disodium hydrogen phosphate (Na2HPO4) and 0.24g of potassium dihydrogen phosphate (KH2PO4) were dissolved in 800ml distilled water. The pH was adjusted to 6.3 using 1N HCl and make up the volume to 1000ml with distilled water.

·Test solution-0.45ml of bovine serum albumin and 0.05ml of test solution of various concentrations were prepared.

·Test control solution-0.45ml of bovine serum albumin and 0.05ml of distilled water were prepared.

·Product control solution -0.45ml of distilled water and 0.05 ml of test solution.

Procedure:

0.05 ml of 250 mg/ml concentrations of standard drug diclofenac sodium were taken respectively and 0.45 ml (0.5% w/V BSA) mixed. The samples were incubated at 37°c for 20 minutes and the temperature was increased to keep the samples at 57°C for 3 minutes. After cooling, add 2.5 ml of phosphate buffer to the above solutions. The absorbance was measured using UV-Visible spectrophotometer at 255 nm. The control represents percentage yield was 3.6% (w/w). The results were compared with Diclofenac sodium. The percentage inhibition of protein denaturation can be calculated as.

Percentage Inhibition = 100 – [{(optical density of test solution – optical Density of control)/optical density of test}/ × 100]

RESULT AND DISCUSSION:

Anti Arthritic activity of Njavara

Medicinal Rice varieties have been of use for age long remedies for human diseases because they contain more pigments and phytochemical components of therapeutic value. Some of them are also used for prophylactic purposes. Medicinal rice is considered to be a chemical factory as it contains multitude of phytochemical compounds like Alkaloids, Flavonoids, Phenol, Glycosides, Saponins, Sesquiterpenes and Oils.

The invitro Anti Artthritic activity was analysed by BSA method. The sample Njavara rice extract showed 31.3, 43.6, 64.3, 77.6 % inhibition of denaturation of bovin serum at the concentration of 100, 250, 500 and 1000 µl whereas standard diclofenac at 250µl showed 51.7% inhibition of denaturation activities (Table 1).

Some literature reported that denaturation of protein is one of the cause of rheumatoid arthritis^8.9. Production of auto-antigens in certain rheumatic diseases may be due to in vivo denaturation of proteins. Mechanism of denaturation probably involves alteration in electrostatic, hydrogen, hydrophobic and disulphide bonding. In our present study, ethanolic extract of Oryza sativa (Njavara) possess anti arthritic activity due the presence of high amount of bioactive compounds especially flavonoids.

Table 1: In-vitro Anti Arthritic Activity of Njavara Rice Extract

S. No	Concentration of Sample	% of Inhibition of Denaturation
1	Standard	51.7 ±3.74
2	100µl	31.3±1.53
3	250 µl	43.6±1.01
4	500 µl	64.3±2.66
5	1000 µl	77.6±1.96

*Values (Mean ± SD) are presented in comparison to standard

CONCLUSION:

In recent years there is an upsurge in the areas related to newer developments in prevention of disease especially Arthritis. So it will be pertinent to examine the possible role of “Anti Arthritic” in its prevention. This study indicates that the Njavara possess the Anti Arthritic Activity. The further study will be carried out for the preparation of medicine for Anti Arthritic and Bone related diseases using the bioactive compound from this selected rice Njavara.

REFERENCE:

1. Singh M, Soni P, Upmanyu N, Shivhare Y. In - vitro Anti - arthritic Activity of Manilkara zapota Linn. Asian J Pharm Tech 2011:1(4);123 -24.

2. Pandey S. Various techniques for the evaluation of anti - arthritic activity in animal models. 2010. J. Adv. Pharm. Tech. Res. 1(2):164 - 170.

3. Tripathi KD. Essentials of Medical Pharmacology. 6th ed. New Delhi:Jaypee Brother’ s Medical Publishers (P) Ltd.; 2008.

4. Bennett PN, Brown MJ. Clinical pharmacology. New Delhi: Churchill Livingstone; 2005.

5. Das GK, Oudhia P. Rice as a medicinal plant inChhattisgarh. NBPGR Newsletter, India 2000, 122:46.

6. Mohanty R. B., Panda, Tripathy B. K 2012. Medicinal rice varieties of India need urgent attention. Current Science, Vol. 102, No. 4, 25.

7. Agrawal S S, and Paridhavi M. Herbal drug technology. University press Pvt. Ltd., Hyderabad, 2007, pp .2.

8. Rahman, H., C.M. Eswaraiah, K. Vakati and P. Madhavi, 2012. In-Vitro Studies Suggest Probable Mechanism of Eucalyptus Oil For Anti-Inflammatory And Anti-Arthritic Activity. International Journal of Phytopharmacy, 2(3): 81-85.

9. Vane, J. R. and R.M. Botting, 1995. New insights into the mode of action of anti-inflammatory drugs. Inflammation Research, 44(1): 1-10.

Received on 10.07.2016 Accepted on 12.10.2016

Asian J. Pharm. Res. 2016; 6(4): 250-252.

DOI: 10.5958/2231-5691.2016.00035.6