Determination of the Mutagenicity Potential of Sankol Herbal Medicine by Single Cell Gel Electrophoresis in Rat Hepatocytes in Comparison With H2O2

Abstract

A B S T R A C T Background:The increasing use of herbal drugs and their ease of accessibility and availability have necessitated the use of mutagenicity test to analyze their toxicity and safety. Objectives: This study aimed to evaluate the genotoxicity of Sankol herbal medicine in DNA breakage of rat hepatocytes in comparison with H 2 O 2 by single cell gel electrophoresis technique or comet assay. Materials and Methods: In the current study hepatocytes were prepared from male wistar rats. Hepatocytes cells were counted and kept in a bioreactor for 30 minutes,then cells were exposed to Sankol herbal medicine at doses of 100, 200 and 400 µl/ml. Buffer 4 (incubation buffer) and H 2 O 2 were used for one hour as negative and positive control respectively. After 30 minutes cell suspension with low melting point agarose was put on precoated slides and covered with agarose gel. Then lysing, electrophoresis, neutralization and staining were carried out. Finally the slides were analyzed by fluorescence microscope. The parameter under this analysis was the type of migration which was determined according to Kobayashi pattern. Results: Results of the study indicated that by increasing the dose of Sankol herbal medicine, the DNA damage slightly increased (P < 0001). Conclusions:In overall compared to the positive control, significant differences were observed which indicated that the crude extract of Sankol in vitro did not have mutagenic effect.