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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 effects of beta-cypermethrin on the growth of meadow grass were studied by MTT assay, microscopy and transmission electron microscopy, fluorescence spectroscopy of single dansyl pentamethylenediamine (MDC) and Western blotting (WB) The cell viability, the characteristics of autophagy and the changes of autophagy-related proteins in Spodoptera frugiperda ovarian cell line Sf9 were tested and analyzed. The results showed that the inhibitory effect of different concentrations of beta-cypermethrin on the activity of Sf9 cells was dependent on the dose and time, and showed typical autophagy. The expression of LC3-Ⅱ / LC3-Ⅰ Level and Beclin 1 protein expression showed an upward trend, respectively 201.32% and 136.00% of the control group, p62 protein expression was down to 68.13% of the control group. Studies have shown that beta-cypermethrin on Spodoptera frugiperda Sf9 cells have significant cytotoxicity, and its inhibition of cell viability and autophagy-induced pathway.