engleski

Evaluation of apoptotic activity of mercury chloride by DNA diffusion assay

Mercury, one of the most widely diffused and hazardous organ-specific environmental contaminants, exists in a wide variety of physical and chemical states, each of them with unique characteristics of target organ specificity. An important mechanism of mercury genotoxicity is its ability to produce free radicals that can cause DNA damage. In our study we tried to find out if those effects are capable of triggering the apoptotic pathway. We used a relatively new method in the detection of apoptosis - DNA diffusion assay. It is based on the diffusion of nucleosomal-sized DNA fragments into the agarose, which gives a hazy or undefined outline without any clear boundary to the apoptotic cell nuclei. The assay was performed on the human whole blood samples. Samples were incubated in 10, 50, 100 and 200 μ M mercury chloride solution at 370 C for 24 and 48 hours. For each treatment protocol a thousand micrographs were analyzed. A significant increase in the apoptosis induction capacity of mercury chloride was observed already after 24 hours of treatment for all tested concentrations. However, no strong correlation between concentration and number of apoptotic micrographs was observed. The correlation was found after 48 hours of treatment but only for doses of 50, 100, and 200 μ M. Based on these results it could be suggested that apoptosis induction is not primary effect of mercury chloride on human lymphocytes. It also supports the theory of mercury’ s organ specificity.

mercury chloride; DNA diffusion assay

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