In this study, the oxidative damage of α-terthienyl (α-T) to the Spodoptera litura (SL) cell and its mechanism were investigated. MTT was used to compare the toxicity of α-T and rotenone to the SL cell. The out...In this study, the oxidative damage of α-terthienyl (α-T) to the Spodoptera litura (SL) cell and its mechanism were investigated. MTT was used to compare the toxicity of α-T and rotenone to the SL cell. The output of malondialdehyde and relative content of glutathione were determined with 2-thiobarbituric acid (TBA) and 5, 5'-dithio-bis (2-nitrobenzoic acid) (DTNB), respectively. Transmission electron microscope (TEM) was employed to observe the influence of α-T on the membrane and organelle of the SL cell. The result showed that the IC50 value of α-T to the SL cell was 0.21 μg mL^-1, whereas the corresponding dose of rotenone was 12.25 μg mL^-1. The output of MDA had the same changing tendency with the concentration of α-T, whereas the content of GSH had the negative correlation with it. According to TEM, the cell membrane and karyotheca swelled and couldn't retain integrity, the intracellular substances leaked out, unidentified granules appeared in the SL cell. The mitochondria expanded, and the membrane and subcellular organelle were damaged severely. In this study, it was found that after oxidative damage induced by α-T, the output of MDA increased notably, whereas the relative content of GSH decreased. This indicated that the antioxidant ability of cell weakened. The result of TEM implied that the SL cell suffered from oxidative damage under the appointed dose.展开更多
Microscopic examination of Spodoptera litura(SL-1)cell infected with AcMNPV or SfaMNPV revealed progressive cell blebbing starting at 8h~12h postinfection and culminating in total cell destruction at 24h postinfectio...Microscopic examination of Spodoptera litura(SL-1)cell infected with AcMNPV or SfaMNPV revealed progressive cell blebbing starting at 8h~12h postinfection and culminating in total cell destruction at 24h postinfection.The fragmentation of the infected cell nuclei was observed by stained with the specific fluorescent dye DAPI.Agarose gel electrophoresis analysis of the DNA extracted from infected cells show typical DNA ladder. All these supported that both viruses infected SL-1 cells undergo apoptosis.Through lipofectin transinfection,we observed that only the DNA of AcMNPV or SfaMNPV can also induce apopotosis of SL-1 cell.Virus titer analysis revealed that neither viruses can duplicate in SL-1 cells.展开更多
Mitoehondria are involved in apoptosis of mammalian cells and even singlecell organisms, but mitochondria are not required in apoptosis in cultured Drosophila cells such as S2 and BG2 cell lines. It is not very clear ...Mitoehondria are involved in apoptosis of mammalian cells and even singlecell organisms, but mitochondria are not required in apoptosis in cultured Drosophila cells such as S2 and BG2 cell lines. It is not very clear whether mitochondria are involved in apoptosis in other insect cells such as lepidopteran cell lines. Thus, we determined to elucidate the role of mitochondria in apoptosis induced by ultraviolet radiation in Spodoptera litura (Lepidoptera: Noctuidae) cell line (SL-ZSU-1). The Western blot results suggested that cytochrome c in the ultraviolet-treated SL-1 cells was released from the mitochondria to cytosol as early as 4 h after the induction of ultraviolet radiation and increased in the cytosolic fractions in a time-dependent manner. Flow cytometric analysis of mitochondrial membrane potential (△ ψm) of SL-ZSU-1 cell treated with ultraviolet-C (UV-C) light indicated the decrease in mitochondrial membrane potential was dependent on the times of ultraviolet treatment. Both of them are different from apoptosis in cultured Drosophila melanogaster cell lines (S2 and BG2) and it appears evident mitochondria are involved in apoptosis of the studied lepidopteran cells.展开更多
基金Natural Science Foundation of Guangdong Province, China (36837) Financial Expenditure Item of Guangdong Province, China (2003C20520) Post-doctoral Science Fund of China (2005037586) for providing the financial support.
文摘In this study, the oxidative damage of α-terthienyl (α-T) to the Spodoptera litura (SL) cell and its mechanism were investigated. MTT was used to compare the toxicity of α-T and rotenone to the SL cell. The output of malondialdehyde and relative content of glutathione were determined with 2-thiobarbituric acid (TBA) and 5, 5'-dithio-bis (2-nitrobenzoic acid) (DTNB), respectively. Transmission electron microscope (TEM) was employed to observe the influence of α-T on the membrane and organelle of the SL cell. The result showed that the IC50 value of α-T to the SL cell was 0.21 μg mL^-1, whereas the corresponding dose of rotenone was 12.25 μg mL^-1. The output of MDA had the same changing tendency with the concentration of α-T, whereas the content of GSH had the negative correlation with it. According to TEM, the cell membrane and karyotheca swelled and couldn't retain integrity, the intracellular substances leaked out, unidentified granules appeared in the SL cell. The mitochondria expanded, and the membrane and subcellular organelle were damaged severely. In this study, it was found that after oxidative damage induced by α-T, the output of MDA increased notably, whereas the relative content of GSH decreased. This indicated that the antioxidant ability of cell weakened. The result of TEM implied that the SL cell suffered from oxidative damage under the appointed dose.
文摘Microscopic examination of Spodoptera litura(SL-1)cell infected with AcMNPV or SfaMNPV revealed progressive cell blebbing starting at 8h~12h postinfection and culminating in total cell destruction at 24h postinfection.The fragmentation of the infected cell nuclei was observed by stained with the specific fluorescent dye DAPI.Agarose gel electrophoresis analysis of the DNA extracted from infected cells show typical DNA ladder. All these supported that both viruses infected SL-1 cells undergo apoptosis.Through lipofectin transinfection,we observed that only the DNA of AcMNPV or SfaMNPV can also induce apopotosis of SL-1 cell.Virus titer analysis revealed that neither viruses can duplicate in SL-1 cells.
基金Acknowledgments This work was supported by the National Natural Science Foundation of China (Grant No: 30470073) and the Natural Science Foundation of Hubei Province (Grant No: 2007ABA 160).
文摘Mitoehondria are involved in apoptosis of mammalian cells and even singlecell organisms, but mitochondria are not required in apoptosis in cultured Drosophila cells such as S2 and BG2 cell lines. It is not very clear whether mitochondria are involved in apoptosis in other insect cells such as lepidopteran cell lines. Thus, we determined to elucidate the role of mitochondria in apoptosis induced by ultraviolet radiation in Spodoptera litura (Lepidoptera: Noctuidae) cell line (SL-ZSU-1). The Western blot results suggested that cytochrome c in the ultraviolet-treated SL-1 cells was released from the mitochondria to cytosol as early as 4 h after the induction of ultraviolet radiation and increased in the cytosolic fractions in a time-dependent manner. Flow cytometric analysis of mitochondrial membrane potential (△ ψm) of SL-ZSU-1 cell treated with ultraviolet-C (UV-C) light indicated the decrease in mitochondrial membrane potential was dependent on the times of ultraviolet treatment. Both of them are different from apoptosis in cultured Drosophila melanogaster cell lines (S2 and BG2) and it appears evident mitochondria are involved in apoptosis of the studied lepidopteran cells.