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Determination of genotoxicity of aqueous extracts of Flueggea leucopyrus willd. (Katupila) using the optimized “Alkaline Comet Assay”

Authors:

R. P. N. H. Rajapaksha,

University of Colombo, LK
About R. P. N. H.
Department of Chemistry, Faculty of Science
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C. D. Wijayarathna

University of Colombo, LK
About C. D.
Department of Chemistry, Faculty of Science
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Abstract

Purpose: The comet assay is a simple, rapid, versatile, sensitive and cost effective method used to measure DNA damage caused by a suspected genotoxic agent. In this study, genotoxic potential of Flueggea leucopyrus Willd., which is used in traditional medicine to treat diseases, including cancers was evaluated using the optimized alkaline comet assay.

 

Methods: Aqueous extracts of Flueggea leucopyrus Willd. leaves (from two different samples isolated from Anuradhapura (FLA) and Maharagama (FLM) was used to evaluate genotoxicity on White Blood Cells (WBC) and Hepato cellular carcinoma cell line (HepG2) cells, using the comet assay. The parameter used to measure the level of genotoxicity in the above cell types was the % tail DNA after treatment with aqueous extract of Flueggea leucopyrus Willd. Different concentrations of FLA and FLM samples (0.001 mg ml-1, 0.005 mg ml-1, 0.01 mg ml-1, 0.02 mg ml-1) were prepared and the effect on WBC and HepG2 cells was evaluated separately, and compared to a negative control (HBSS for WBC and culture media for HepG2) and positive control (200 μM H2O2 for WBC and 50 μM H2O2 for HepG2). The results were analyzed by one way ANOVA followed by Dennett post hoc test using SPSS software.

Results: As the concentration of the aqueous extract was increased, the % tail DNA of both WBC and HepG2 cells increased gradually, compared to the negative controls (p<0.05).

Conclusion: Aqueous extracts of Flueggea leucopyrus Willd. appear to be moderately genotoxic at high concentrations, but the effect is insignificant at lower concentrations.

How to Cite: Rajapaksha, R.P.N.H. & Wijayarathna, C.D., (2017). Determination of genotoxicity of aqueous extracts of Flueggea leucopyrus willd. (Katupila) using the optimized “Alkaline Comet Assay”. Pharmaceutical Journal of Sri Lanka. 7, pp.23–33. DOI: http://doi.org/10.4038/pjsl.v7i0.21
Published on 08 Aug 2017.
Peer Reviewed

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