Evaluation of In Vitro Anti Inflammatory activity of Kusta Gaja Kesari - A Siddha Herbo Mineral Formulation against Albumin Protein Denaturation

 

A. Sureka^1*, C. Mary Sharmila², R. Chithra Devi ³, N. J. Muthu Kumar ⁴, V. Banumathi⁵

¹Emergency Medical Officer, National Institute of Siddha, Tambaram Sanatorium.

²Resident Medical Officer, National Institute of Siddha, Tambaram Sanatorium.

³House Officer, National Institute of Siddha, Tambaram Sanatorium.

⁴Associate Professor, Hospital Superintendent, National Institute of Siddha, Tambaram Sanatorium.

⁵Director, National Institute of Siddha, Tambaram Sanatorium.

 

ABSTRACT:

Aim: The present study is conducted to evaluate the In Vitro Anti Inflammatory activity of Kusta Gaja Kesari (KGK) against Protein (Albumin) Denaturation. Methods: The test drug (KGK) at varying concentration ranges from 100 to 500 µg/ml is incubated and heated with Egg albumin in controlled experimental conditions. The percentage inhibition of the protein denaturation is calculated. Diclofenac sodium is used as reference standard drug. Result: The results from the present study clearly indicates that the test drug KGK is effective in inhibiting heat induced protein denaturation. A maximum percentage of inhibition of 51.21 % was observed at 500 μg/ml when compared with the standard.

 

 

 

INTRODUCTION:

Inflammation is defined as the local response of living mammalian tissues to injury caused due to any agent. It is a body defence reaction in order to eliminate or limit the spread of injurious agent, followed by removal of the necrosed cells and tissues. Inflammation is a protective response by the body to variety of etiological agents may be Infectious or Non–Infectious¹. Inflammation is a more complex process than what is understood.

 

 

It is diverse and is now recognised as the underlying pathologic process involved in the onset of almost all pathologies ranging from a acute inflammation in the skin due to a infection to the chronic inflammation resulting in remodelling of the arterial wall in atherosclerosis². The cardinal signs of Inflammation were first given by the Roman writer celsus in 1^st century A.D as Rubor, tumor, calor, and dolor. To these the fifth sign function laesa (loss of function) was latter added by Virchow¹. Inflammation is the response of the immune system to any damage to tissues and due to infection. The immune system plays an important role in the maintaining the health of all cells of the body. Focussing on research in inflammation , involves not only the study of immunological and cellular responses but also the pharmacological processes involved in drug development ³.

 

Kusta Gaja Kesari is a siddha herbo mineral formulation mentioned in the Siddha Text “Siddha Vaithiya Thirattu. It is indicated for the treatment of various skin diseases like leprosy, leucoderma and chronic skin diseases. In search of effective traditional drugs to target inflammation, the author has made an attempt to evaluate the anti-inflammatory activity of Kusta Gaja Kesari, a drug with traditional properties to target the inflammatory reactions in the skin.

 

MATERIALS AND METHODS:

Kusta Gaja Kesari consists of Abraga chenduram, Kaantha Chenduram, Aya chenduram, Rasa Parpam prepared with fruit extracts of Savuri Pazham (Trichosanthes tricuspidata). KGK was obtained from IMPCOPS pharmacy; Chennai, Tamilnadu.

 

Albumin Denaturation Assay Procedure:

The in-vitro anti-inflammatory activity KGK was studied using albumin denaturation technique. The reaction mixture consisted of bovine serum albumin (5% aqueous solution) and test sample KGK at varying concentration ranges from 100 to 500 µg/ml and the standard Diclofenac sodium at a concentration of 100 µg /ml of final volume. The pH was adjusted by using a small amount of 1N Hydrochloric acid. The samples were incubated at 37°C for 20 min and then heated at 57°C for 3 min. After cooling the sample, 2.5 ml of phosphate buffer solution was added into each test tube. The Turbidity developed was measured spectrophotometrically at 660 nm, for control distilled water was used, as the product control tests lacked bovine serum albumin. The experiment was performed in triplicate.

 

The Percentage protection from denaturation was calculated by using the below given formulae ^4,5.

 

^(A) Control - ^(A) sample

----------------------------------------------x100

^(A) Control

Statistical analysis:

Results are expressed as Mean ± SD. The difference between the experimental groups was compared by One-Way Analysis Of Variance (ANOVA) followed by Dunnet Multiple comparison test.

 

 

Figure 1 - Preparation of Test and control

 

Figure 2 - Absorbance of reaction mixture – Test Sample

 

 

Figure 3 - Absorbance of reaction mixture – Control and Standard

 

 

Figure 4 - Absorbance Range of test and standard at Trial 1

 

 

Figure 5 - Absorbance Range of test and standard at Trial 2

 

 

Figure 6 - Absorbance Range of test and standard at Trial 3

 

Table 1 – Percentage Inhibition of Protein Denaturation

Concentration in µg/ml	Percentage Inhibition of Protein Denaturation
KGK 100	10.62 ± 7.39
KGK 200	18.8 ± 8.51
KGK 300	28.79 ± 6.34
KGK 400	35.23 ± 5.62
KGK 500	51.21 ± 9.02
Diclofenac sodium (100 µg)	91.93 ± 1.56

Each value represents the mean ± SD. N=3

 

RESULT AND DISCUSSION:

The results obtained from the present study clearly indicates that the test drug KGK was effective in inhibiting the heat induced albumin denaturation. The results for the various concentration of the test drug KGK is given in Table 1 and the absorbance range of the test drug and the standard are given in Figure 4, Figure 5, Figure 6. Maximum percentage inhibition of about 51.21 % was observed at 500 μg/ml of the test drug Kusta Gaja Kesari when compared to that of the Diclofenac sodium, a standard anti-inflammatory agent with the maximum inhibition 91.93 % at the concentration of 100 μg/ml. The study is a preliminary attempt to establish the anti inflammatory activity of Siddha Medicinal Formulation Kusta Gaja Kesari. Further studies are needed to widen and substantiate the therapeutic utility of the drug.

 

CONCLUSION:

From the result of the study it can be concluded that the test drug Kusta Gaja Kesari (KGK) possess promising anti-inflammatory property in protein denaturation assay.

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

 

REFERENCE:

1.        Harsh Mohan, Pathology quick review and MCAS 3^rd edition, Jaypee Brothers Medical Publishers (p) Ltd, Chapter 6, Inflamation &Healing, 2010, 92.

2.        Neville A Punchard, Cliff J Whelan and Ian Adcock. The journal of inflammation (Lond), 2004;1(1).

3.        www.cell.com/trends/Molecular-medicine.

4.        G.Leelaprakash, S.MohanDass. In-vitro anti-inflammatory activity of methanol extract of enicostemmaaxillare. Int. J. Drug Dev. & Res., 2011, 3 (3): 189-196.

5.        M. V. Anoop, A. R. Bindu . In-vitro Anti-inflammatory Activity Studies on Syzygiumzeylanicum (L.) DC Leaves. International Journal of Pharma Research & Review, August 2015; 4(8):18-27.

 

 

 

 

Received on 08.06.2018          Accepted on 16.07.2018        

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Res. 2018; 8(3):  145-147.

DOI: 10.5958/2231-5691.2018.00025.4