The DNA Damage And Apoptosis Induced By Spinosad And Chlorpyrifos

Nowadays the changes of pesticide are mainly due to the environmental requirements. Modernized insecticides get a lot of attention for their safety to environment and human health, while they can control pests efficiently. Spinosad and chlorpyrifos are high-efficient pesticides widely used in agricultural production. Although there are already a lot reports about spinosad and chlorpyrifos, but the accurate mechanism of spinosad is still unclear. Especially the research about in vitro genotoxic effects on spinosad and chlorpyrifos is rare. In this paper, we took three mammalian cell lines Hela, HepG2, Hek293and three insect cell lines Tn5B1-4, Sf9, S2as the object of study to investigate and analyze the cytophysiological index including DNA damage, mitochondrial membrane potential and apoptosis induced by spinosad and chlorpyrifos through the means of single cell gel electrophoresis, yH2AX foci formation and flow cytometry, which preliminarily elucidated the genotoxic mechanism of spinosad and chlorpyrifos.Study indicated that different cell lines treated with spinosad and chlorpyrifos respectively, the cell viability declined following the increase of drug concentration and treatment time. Relatively speaking, mammalian cells,Hela, HepG2and Hek293are sensitive to spinosad, insect cell Tn5Bl-4is more sensitive to chlorpyrifos than HepG2cell. High concentration of spinosad and chlorpyrifos would cause obvious comet image, the tail length, tail area and the number of cells had comet tail grew with the concentration of insecticides. The number of yH2AX foci and cells formed yH2AX foci increased following the increase of concentration of spinosad and chlorpyrifos. The results of flow cytometry showed that spinosad and chlorpyrifos could induce the apoptosis of mammalian cells, the rate of apoptosis increased with increasing of drug concentration. At the same time, these two insecticides could induce the decrease of mitochondrial membrane potential, it also suggested that insecticides could cause early apoptosis. Through AO-EB double staining, we found that spinosad and chlorpyrifos treated Tn5Bl-4, Sf9and S2cell lines, apoptosis occurred, the percentage of apoptotic cells grew with the increase of insecticides concentration. The conclusions suggested that spinosad and chlorpyrifos could induce DNA damage and apoptosis of mammalian cells,Hela, HepG2, Hek293and insect cells Tn5Bl-4, Sf9, S2. These two insecticides had obvious genotoxicity.