ANTI-INFLAMMATORY AND ANTIBACTERIAL STUDIES ON INDIGOFERA LINIFOLIA RETZ.

ANTI-INFLAMMATORY AND ANTIBACTERIAL STUDIES ON INDIGOFERA LINIFOLIA RETZ.

M. Uvarani*, R. Arun Pandian, B. Jenila, R. Sutharsingh and A. Thangathirupathi

Department of Pharmacognosy, Sankaralingam Bhuvaneswari College of Pharmacy, Anaikuttam, Sivakasi-626130,Tamilnadu, India

ABSTRACT

Indigofera linifolia Retz(Fabaceae) is herb which spread throughout the India and found to be used by the tribal to cure inflammations. Based upon the review the present study focused on screening of Ant-inflammatory and Anti-bacterial activity of Indigofera linifolia Retz. The coarse powder of whole plant was extracted by cold maceration method using various solvents successively. Preliminary phytochemical screening was carried out to confirm the presence of phytoconstituents. Based upon the phytochemical tests chloroform and ethanol extracts (200mg/kg) were taken for anti-inflammatory and antibacterial activity screening. Anti inflammatory studies was carried out by Carrageenan induced paw edema method in Wistar albino rats. Anti bacterial screening was carried out in chloroform and ethanolic extract by disc diffusion method in three different concentrations (100μg, 150μg, 200μg /disc) and the diameter of the zone of inhibition were recorded. The percentage increase in paw edema was reduced in both chloroform (18.20±1.32) and ethanol extract (24.22±1.12) treated animals when compared with standard Indomethacin (19.18±1.02) and control (40.10±1.22) animals. The chloroform extract possessed significant anti inflammatory activity that may be due to its ability to prevent the production of some pro-inflammatory mediators. The chloroform and ethanolic extract showed the inhibitory activity against all the four strains of organisms , but the ethanolic extract showed potent inhibition against Streptococcus pneumoniae and Staphylococcus aureus in higher concentration (200 μg/disc) when compared with Ciprofloxacin (10µg /disc).

Kirby-Bauer disc diffusion method.

INTRODUCTION: Indigofera linifolia Retz (Fabaceae) is herb which spread throughout the India and found to be used by the tribal for various ailments. The leaves used as antibacterial, anti-oxidant and also used for its cytotoxicity effect in lung cancer. From the ethno botanical survey known that the entire plant was specifically used to cure inflammations ^1-6. Based upon the review the present study focused on screening of Ant-inflammatory and Anti-bacterial activity of Indigofera linifolia Retz.

FIGURE 1: ENTIRE PLANT OF INDIGOFERA LINIFOLIA RETZ.

MATERIALS AND METHODS:

Collection and Extraction: The plant was authenticated and collected from Anaikuttam dam, near Sivakasi, Tamilnadu. The coarse powder was extracted by cold maceration method using various solvents such as hexane, chloroform, ethyl acetate and ethanol successively by cold maceration method. All the extracts were concentrated under reduced pressure and subjected to preliminary phytochemical screening ^7-8.

Acute Oral Toxicity: Acute oral toxicity was performed as per OECD guideline 423 in Swiss mice. The animal studies were carried out with the approval of institutional animal ethical committee. The chloroform and ethanolic extracts were administered orally in a single dose (2000 mg/kg) by gavage. Animals were observed for14 days for the symptoms such as tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma were observed.

Anti-inflammatory Activity: The anti-inflammatory study was carried out by Carrageenan induced paw edema method in Wistar albino rats ^9-10. The animals were divided into four groups such as chloroform extract (200mg/kg) and ethanol extract (200mg/kg), standard indomethacin (10mg/kg) and vehicle control groups. The paw volume was recorded at 0, 1, 2, 3 and 4 hr after carrageenan injection using a volume transducer attached with strain gauge coupler of student physiograph. The percentage increases in paw volume were calculated from the 4th hr reading.

TABLE 1: EFFECT OF EXTRACTS ON CARRAGEENAN INDUCED PAW EDEMA IN RATS

Values are expressed as mean ± SEM. **p<0.01 (One-way ANOVA followed by Dunnett’s test)

Antibacterial Screening: Antibacterial screening was carried out by Kirby-Bauer disc diffusion method in chloroform and ethanolic extract using three different concentrations (100μg, 150μg, 200μg /disc) respectively. Both chloroform and methanol extracts were dissolved in dimethyl sulfoxide (DMSO). The strains of Escherichia coli (ATCC No. 8739), Salmonella typhi (ATCC 13311), Streptococcus pneumonia (ATCC 33400) and Staphylococcus aureus (ATCC 25923) were used. The Ciprofloxacin antibiotic disc (10µg /disc) was used as standard. About 25ml of Muller-Hinton Agar media was transferred into four sterile petridish. Each petridish were inoculated with 0.2ml of each inoculum by using sterile cotton swap. Sterile paper disc, impregnated with different concentrations of chloroform and ethanolic extracts were placed on the surface of the medium in each petri dish. The plates were kept in incubator overnight and the zone of inhibition diameter was measured ^11-13.

RESULTS: From the phytochemical study the presence of steroids, phenolics, saponins and flavonoids were identified in chloroform and ethanolic extracts of Indigofera linifolia. In the anti-inflammatory studies, the percentage increase in paw edema was reduced in both chloroform and ethanol extract treated animals when compared with standard Indomethacin. The chloroform and ethanolic extract showed the inhibitory activity against all the four strains of organisms. The ethanolic extract showed potent inhibition against Escherichia coli, Salmonella typhi, and Staphylococcus aureus in higher concentration (200μg/disc) when compared with Ciprofloxacin (Table 2, fig. 2-5).

TABLE 2:  ZONE OF INHIBITION IN DISC DIFFUSION METHOD

Values are expressed as mean ± SEM. N = 3

FIGURE 2: INHIBITORY EFFECT OF INDIGOFERA LINIFOLIA EXTRACTS AGAINST STREPTOCOCCUS PNEUMONIAE

FIGURE 3: INHIBITORY EFFECT OF INDIGOFERA LINIFOLIA EXTRACTS AGAINST ESCHERICHIA COLI

FIGURE 4: INHIBITORY EFFECT OF INDIGOFERA LINIFOLIA EXTRACTS AGAINST SALMONELLA TYPHI

FIGURE 5: INHIBITORY EFFECT OF INDIGOFERA LINIFOLIA EXTRACTS AGAINST STAPHYLOCOCCUS AUREUS

DISCUSSION: The chloroform extract possessed significant anti inflammatory activity. Activity may be due to its ability to prevent the production of some pro-inflammatory mediators. From the anti bacterial screening both extract showed moderate inhibition in the all the test organism.

REFERENCE

1. Renukadevi K.P., Suhani Sultana, International Journal of Pharmacology, 2011:7(3); Page No: 9-17.
2. Antony Dweck C., International Journal of Cosmetic Sciences, 2002: 24(5); 287-302.
3. Muhammad Nisar, Shafiq Ahamed Tariq, Ihasaullah, 2005:  Pakistan journal of Botany, 41(3): 1359-1361.
4. Tamilselvi.N., Dhamodharan R., Krisknamoorthi P., Shivakumar, J.Chem.Pharm.Res., 2011: 3(2) ; 738-746.
5. Flora of China, Vol. 10, 137-143.
6. Nadkarni A.k., Indian Material Medica., Vol. 1; 680.
7. Khandelwal K.R., Practical Pharmacognosy, Techniques and Experiments, (13th Edn.) Nirali Prakashan, Pune, India, 2006, 30-76.
8. Anees Ahmad Siddiqui and Mohammed Ali, Practical Pharmaceutical Chemistry, 1st edition, CBS Publishers and Distributors, New Delhi,1997,126-132.
9. Agarwal RB and Rangari VD. Anti-inflammatory and Anti-arthritic activities of Lupeol and 19a-H Lupeol isolated from Strobilanthus callosus and Strobilanthus ixiocephala roots. Indian Journal of Pharmacology. 2003; 35: 384-387.
10. Kamalutheen M, Gopalakrishnan S and Syed Ismail T. Anti-inflammatory and Anti-arthritic Activities of Merremia tridentata (L.) Hall. E-Journal of Chemistry. 2009; 6(4): 943-948.
11. Karaman I., Sahin F., Gulluce M., Ogutcu H., Sengul M. and Adiguel A., Antimicrobial activity of aqueous and methanol extracts of Juniperous oxycedrus L., Journal of Ethnopharmacology, 2003; 85: 231-235.
12. Cruikshank R., Duguid J.P., Marmion B.P. and Swain R.H., Medical Microbiology, Part-II, (12th edition) Churchill Livingston Publications, Edinburg, 1975, 120.
13. Oyeleke, S. B., Dauda, B. E. and Boye, O. A., Antibacterial activity of Ficus capensis, African Journal of Biotechnology, 2008; 7 (10): 1414-1417.
    

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    How to cite this article:

    Uvarani M, Pandian RA, Jenila b, Sutharsingh R and Thangathirupathi A. Anti-Inflammatory and Antibacterial Studies on Indigofera linifolia Retz.. Int J Pharm Sci Res 2012; Vol. 3(8): 2754-2756.

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