Determination of the Mutagenicity Potential of Dillsun Herbal Medicine by Single Cell Gel Electrophoresis in Rat Hepatocytes

BACKGROUND: Traditional medicines are among the oldest medicines and their extensive use in the recent years reflects the public’s interest in alternatives to conventional medicine. OBJECTIVES: The aim of this study was to investigate the genotoxicity of Dillsun herbal medicine in DNA damage of rat hepatocytes compared to sodium dichromate using a comet assay technique. MATERIALS AND METHODS: Male Wistar rats were caught and their liver was washed with a perfusion buffer, followed by another wash with collagenase buffer. Hepatocytes were isolated and transferred on to a petri dish which contained a washing buffer. Hepatocytes were then separated and the cells were filtered and centrifuged at 1500 rpm for 3 minutes. The hepatocytes were counted using neubauer slides and kept in a bioreactor for 30 minutes. Cells were then exposed to different doses of Dillsun such 0.2, 1, 2.5, 5 and 10 mg/mL. Sodium dichromate was the positive control and incubated buffer was used as a negative control. Cell suspensions were placed on slides pre-coated with low melting point agarose and were covered with agarose gel. Agarose gels were then lysed and electrophoresis was done, followed by neutralization and staining. Slides were analyzed by fluorescence microscopy. The size and extent of DNA damage visualized by this technique was evaluated by examining cells. Migration behavior was classified according to the Kobayashi pattern. RESULTS: The results indicated that with an increase of Dillsun dose, the mutagenicity index slightly increased but compared to the positive control, there were significant differences, which suggests that the crude extract of Dillsun in vitro did not have mutagenic effects. CONCLUSIONS: In conclusion the results showed that Dillsun has no mutagenic effects when compared to the positive control. Although by increasing the Dillsun dose, DNA damage also increased but this increase was not significant.