Apoptosis Of Cell Induced By Tebufenozide

Insect growth regulator is a novel insecticide,which kills the insects by regulating the growth and development of them.Tebufenozide,a new class of insect growth regulator and non-steroidal ecdysone agonists,killed insects by controlling the molting process.In this paper,we studied the mechanism of cytotoxicity induced by Tebufenozide on mammalian cell lines Hela cells and the insect cell lines Tn5B1-4 cells.The effect of Tebufenozide on the viability of cells was determined by MTT assay,and the results showed that the viabilities of Hela cells and Tn5B1-4 cells were inhibited.And the inhibition of Tn5Bl-4 cells is greater than Hela cells.We determined the apoptosis of Hela cells and Tn5B1-4 cells by flow cytometry and AO/EB staining,and the results suggested Tebufenozide induced the apoptosis of the two cells.From the results of the comet assay and the DAPI staining assay,we found Tebufenozide could induce the DNA damage.According the results of apoptosis,we detected the relative apoptosis pathway.The data of Rhodamine 123 showed Tebufenozide induced the loss of mitochondrial membrane potential.The generation of ROS was increasing induced by Tebufenozide through the DCFH-DA staining assay.We detected the level of cytochrome C and the activation of caspase-3 by western blot assay.The results showed that cytochrome C in cytosol and the activation of caspase-3 was increasing induced by Tebufenozide.At the same time,we pretreated the cells with the anti-oxidant NAC,and studied the effects of NAC on the toxicity of Tebufenozide.From the results we suggested that NAC could decrease the toxicity of Tebufenozide on cells.The conclusions suggested that tebufenozide induced apoptosis of Hela cells and Tn5B1-4 cells through the mitochondrial pathway,and the anti-oxidant NAC could decrease the toxicity.