不同杀虫成分对Sf9细胞凋亡的影响

Effects of Different Insecticidal Components on Induction of Apoptosis in Spodoptera frugiperda Cultured Cell Line Sf9

通过倒置显微形态观察凋亡小体、DNA琼脂糖凝胶电泳确证的模型,研究了8种代表性杀虫成分对草地贪夜蛾Spodoptera frugiperda离体培养细胞系Sf9的凋亡诱导效果,以探讨不同杀虫成分对昆虫细胞凋亡的影响.结果表明,印楝素0.15-1.50μg/mL处理后72 h内、喜树碱0.35-3.50μg/mL处理后36 h,Sf9细胞产生大量典型凋亡小体,处理时间延长或剂量加大则以造成细胞坏死为主;DNA电泳产生典型的DNA Ladder,表明印楝素和喜树碱对Sf9具有明显的细胞凋亡诱导作用.印楝素1.5μg/mL诱导处理Sf9细胞凋亡高峰期为处理后24-48 h,喜树碱0.35-3.50μg/mL诱导处理Sf9细胞凋亡高峰期为处理后12 h.苦参碱、氟铃脲、毒死蜱、灭多威、氯氰菊酯以0.1-10.0μg/mL及昆虫蜕皮激素20-羟基蜕皮酮（20-hydroxyecdysone）以0.048-4.800μg/mL处理后72 h内对Sf9均无明显的细胞凋亡诱导作用.杀虫剂对昆虫细胞凋亡诱导效应随供试化合物、细胞种类、处理剂量、处理时间而异.

The induction of apoptosis of eight typical insecticidal components, including azadirachtin, in Spodoptera frugiperda cultured cell lines Sf9 were determined in this paper to investigated the specificity of apoptosis induction by insecticides through the research model of inverted phase contrast microscope assay and DNA agarose gel electrophoresis. The results were showed that the typical morphological characteristics of apoptosis appeared, including a lot of apoptotic bodies, treated in 72 h with azadirachtin at a mass concentration of 0.15 - 1.50 μg/mL and 36 h with comptothecin at 0. 35 - 3.50 μg/mL. The necrosis would become a dominant role after the treatment prolonged or the concentration increased. The typical DNA Ladder was got by agarose gel electrophoresis, which showed that azadirachtin and comptochetin possessed distinct apoptosis induction and the induction peak time was 24 -48 h or 12 h after treatment with at a mass concentration of 1.5 μg/mL of azadirachtin or 0. 35 - 3.50 μg/mL of comptochetin, respectively. However, the same changes could not appeared in Sf9 cells within 72 h treated at a mass concentration of 0. 1 - 10. 0 μg/mL of matrine, hexaflumuron, chlorpyriphos, methomyl or cypermethrin, and 0. 048 - 4. 800 μg/mL of 20-hydroxyecdysone, respectively. It was suggested that the apoptosis induction by insecticide would be specific and the induction effectiveness depend on the kind of test chemicals, cell line, treated close and time.