高效氯氰菊酯对草地贪夜蛾Sf9细胞自噬的诱导作用

Beta-cypermethrin induced Spodoptera frugiperda Sf9 cells autophagy research

采用噻唑蓝(MTT)染色、显微镜及透射电子显微镜观察、单丹磺酰戊二胺(MDC)荧光染色及蛋白质免疫印迹(WB)技术等方法,分别针对高效氯氰菊酯(beta-cypermethrin)作用后草地贪夜蛾Spodoptera frugiperda卵巢细胞系Sf9细胞的细胞活力、自噬发生的典型特征及自噬相关蛋白变化情况进行了测试及分析。结果表明:经不同浓度高效氯氰菊酯处理后,对草地贪夜蛾Sf9细胞活性的抑制效应与作用剂量及时间呈现依赖关系,并表现出典型的细胞自噬现象;LC3-Ⅱ/LC3-Ⅰ蛋白表达水平比和Beclin 1蛋白表达量呈现上调趋势,分别为对照组的201.32%和136.00%,p62蛋白表达量下调至对照组的68.13%。研究表明,高效氯氰菊酯对草地贪夜蛾Sf9细胞具有显著的细胞毒性,并且其对细胞活力的抑制作用与自噬诱导途径相关。

The cell viability and autophagic vacuoles of the beta-cypermethrin treated Sf9 cells （Spodoptera frugiperda cell） were evaluated and the autophagic-related proteins of Sf9 cells was analyzed. 3-（4,5-Dimethyl-2-thiazolyl）-2, 5-diphenyl-2-H-tetrazolium bromide（MTT） assay, microscope assay, transmission electron microscope（TEM） assay, monodansylcadaverine（MDC） fluorescent staining assay and western blot（WB） assay were employed for the investigation. Results showed that beta-cypermethrin inhibited the viability of Sf9 cells in both concentration- and time- dependent manners, and the autophagy was observed in beta-cypermethrin-treated cells. Autophagy- related protein LC3-Ⅱ/LC3-Ⅰ and Beclin 1 were up-regulated to 201.32% and 136.00% compared to the control group. And protein p62 was down-regulated to 68.13%. It has been demonstrated that beta- cypermethrin has significant cytotoxicity on Sf9 cells and inhibition the cell＇s ability via the autophagy pathway.