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Article in Chinese | WPRIM | ID: wpr-882219


Objective:To establish a comet test method for detection of genotoxicity of three reference chemicals in rat liver cells. Methods:6-10 week old Sprague Dawley rats were randomly divided into 4 groups, with normal saline (0.9% NaCl solution) as negative control group. Animals in three test groups were treated, respectively, with ethyl methanesulfonate (EMS) 200 mg/kg, N-methyl-N-nitrosourea (MNU) 50 mg/kg, and D-mannitol 2 000 mg/kg. There were 10 animals in each group, 5 males and 5 females. The animals received two times (21 h interval) of test compounds through intragastric administration, and their clinical symptoms and body weight changes were recorded during the experiment. The rats were sacrificed 3 h after the last exposure. The liver was weighed, then used to prepare single-cell suspensions for the alkaline comet test which determines the average tail DNA content percentage (DNA%) of hepatocytes and other comet indicators. Results:(1) D-mannitol, EMS and MNU did not show significant toxicity in the whole animal. (2) The mean values of tail DNA content percentage (DNA%) of rat hepatocytes in EMS [(60.07±24.69)%] and MNU [(41.66±22.35)%] groups were higher than that in the negative control group [(2.32±1.39)%] and the difference was statistically significant (P<0.05). The difference between D-mannitol group [(3.06±3.30)%] and the negative control group was not significant (P>0.05). Conclusion:This laboratory has established a comet test method using hepatocytes from treated rats. Among three testing chemicals, EMS and MNU have displayed genotoxicity by this assay, but no genotoxicity was observed in D-mannitol treated animals.