Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China.
CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China.
Comp Biochem Physiol C Toxicol Pharmacol. 2021 Nov;249:109130. doi: 10.1016/j.cbpc.2021.109130. Epub 2021 Jul 8.
Avermectin (AVM), is widely applied in the fields of agriculture, possess activities against mites and insects. AVM is generally thought to keep the GABA-related chloride channels open in insect cells. However, AVM induces cytotoxicity in non-neural cells still ambiguous. Here we evaluate the cytotoxicity and other mode of action of AVM in Spodoptera frugiperda (Sf9) cells. Our results showed that AVM suppressed the activity of Sf9 cells and induced programmed cell death. DNA damage of Sf9 cells was detected by alkaline comet assay and PARP. The cleavage of poly ADP-ribose polymerase (PARP) and DNA double-strand breaks demonstrated AVM induced DNA damage in Sf9 cells. In addition, a series of established cytotoxicity tests were conducted to explore the mechanism of AVM toxicity in Sf9 cells. Typical apoptosis changes were occurred including increasing the expression of Bax/Bcl-2 and the activation of caspase-9/-3. Subsequently, Western blotting was used to detected autophagy related proteins including LC3, Beclin1 and p62. We found that AVM upregulated LC3, Beclin1 expression and downregulated p62 expressions. Moreover, we found that AVM induced autophagy may through AMPK/mTOR-mediated autophagy pathway. These results showed that AVM-induced DNA damage and programmed cell death in Sf9 cells.
阿维菌素（AVM）广泛应用于农业领域，具有抗螨和杀虫活性。AVM 通常被认为能使昆虫细胞中的 GABA 相关氯离子通道保持开放。然而，AVM 在非神经细胞中诱导细胞毒性的机制仍不清楚。在这里，我们评估了 AVM 在草地贪夜蛾（Sf9）细胞中的细胞毒性和其他作用模式。我们的结果表明，AVM 抑制 Sf9 细胞的活性并诱导程序性细胞死亡。通过碱性彗星试验和 PARP 检测 Sf9 细胞的 DNA 损伤。多聚 ADP-核糖聚合酶（PARP）的切割和 DNA 双链断裂表明 AVM 诱导 Sf9 细胞的 DNA 损伤。此外，还进行了一系列既定的细胞毒性试验，以探索 AVM 在 Sf9 细胞中的毒性机制。包括 Bax/Bcl-2 表达增加和 caspase-9/-3 激活在内的典型凋亡变化发生。随后，通过 Western blot 检测自噬相关蛋白，包括 LC3、Beclin1 和 p62。我们发现 AVM 上调了 LC3、Beclin1 的表达，下调了 p62 的表达。此外，我们发现 AVM 诱导的自噬可能通过 AMPK/mTOR 介导的自噬途径。这些结果表明，AVM 诱导 Sf9 细胞中的 DNA 损伤和程序性细胞死亡。
