Natural β-carboline alkaloids regulate the PI3K/Akt/mTOR pathway and induce autophagy in insect Sf9 cells
Natural β-carboline alkaloids regulate the PI3K/Akt/mTOR pathway and induce autophagy in insect Sf9 cells
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天然 β-咔啉生物碱调节 PI3K/Akt/mTOR 通路并诱导昆虫 Sf9 细胞自噬
DOI：
10.1016/j.pestbp.2018.12.005
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发表时间：
2019-02-01
影响因子：
4.7
通讯作者：
Zhong, Guohua
中科院分区：
文献类型：
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作者：
Cui, Gaofeng;Shu, Benshui;Zhong, Guohua
β-咔啉生物碱是一大类天然存在的和合成的吲哚生物碱，具有显著的药理学性质。此外，这些生物碱也被报道是用于控制许多害虫和植物病原线虫的有效药剂。然而，对这些潜在的杀虫成分的研究很少。这些生物活性化合物能够诱导肿瘤细胞程序性死亡的研究为探索其对昆虫的毒理机制提供了新的思路。本研究测定了5种天然骆驼蓬生物碱的细胞毒性，并在草地贪夜蛾Sf 9培养细胞系中证实了其诱导自噬的作用。单丹酰尸胺染色和LysoTracker Red染色显示，不饱和β-咔啉生物碱去氢骆驼蓬碱和去氢骆驼蓬酚具有较强的自噬诱导活性。在RNA水平上，β-咔啉处理后，许多自噬相关基因表达增加，Sf-Atg 8的蛋白表达也被证实在处理后增加。此外，主要的自噬信号通路PI 3 K/Akt/mTOR通路在手术过程中发生了改变。此外，进行了特殊抑制剂和活化剂的实验，以确认β-咔啉对该途径的影响。结果表明，PI 3 K/Akt/mTOR信号通路主要调控骆驼蓬碱诱导的昆虫细胞自噬，这一发现可能有利于这些具有潜在生物活性成分的应用。
The beta-carboline alkaloids are a large group of naturally occurring and synthetic indole alkaloids with remarkable pharmacological properties. Furthermore, these alkaloids have also been reported to be effective agents for controlling many pests and plant pathogenic nematodes. However, studies on these potential insecticidal components are scarce. The previous finding that these bioactive compounds can induce programmed cell death in cancer cell lines provided a new insight for exploration of their toxicological mechanisms on insects. In the present study, the cytotoxicity of five natural harmala alkaloids was measured, and the autophagy-inducing effect was confirmed in the Spodoptera frugiperda Sf9 cultured cell line. The results demonstrated that these alkaloids inhibited the proliferation of Sf9 cells in a dose- and time-dependent manner, and the unsaturated beta-carboline alkaloids, harmine and harmol, exhibited stronger autophagy induction activity based on mono-dansylcadaverineand LysoTracker Red staining. Many autophagy-related genes were increased after beta-carbolines treatment at the RNA level, and the protein expression of Sf-Atg8 was also confirmed to increase after treatment. In addition, the primary autophagic signaling pathway, the PI3K/Akt/mTOR pathway, was altered during the procedure. Furthermore, experiments with special inhibitors and activators were performed to confirm the effect of beta-carbolines on this pathway. The results suggested that the PI3K/Akt/mTOR pathway primarily regulated harmine-induced autophagy in insect cells, and this finding may potentially benefit the application of these promising bioactivity components.