School of Plant Protection, Hainan University, Haikou 570228, China.
Institute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences, Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization, Ministry of Agriculture and Rural Affairs, Guangzhou 510640, China.
Toxins (Basel). 2022 Sep 29;14(10):677. doi: 10.3390/toxins14100677.
Furanocoumarins, the secondary metabolites of plants, are considered to be natural insecticides and fungicides because they prevent the invasion of plant pathogenic microorganisms and the predation of herbivorous insects. In this study, novel 2-arylfuranocoumarin derivatives were designed to synthesize by condensation, esterification, bromination, and Wittig reaction. The results showed an excellent photosensitive activity of 2-thiophenylfuranocoumarin (I34). Cell Counting Kit-8 detected that I34 could inhibit the proliferation of Spodoptera frugiperda (Sf9) cells in a time- and concentration-dependent manner under ultraviolet A (UV-A) light for 3 min. The inverted microscope revealed that cells treated with I34 swelled, the membrane was ruptured, and apoptotic bodies appeared. The flow cytometry detected that I34 could induce apoptosis of Sf9 cells, increase the level of intracellular reactive oxygen species (ROS), decrease the mitochondrial membrane potential, and block cell cycle arrest in the G2/M phase. Transmission electron microscopy detected cell mitochondrial cristae damage, matrix degradation, and mitochondrial vacuolation. Further enzyme activity detection revealed that the enzyme activities of apoptosis-related proteins caspase-3 and caspase-9 increased significantly (p < 0.05). Finally, Western blotting analysis detected that the phosphorylation level of Akt and Bad and the expression of the apoptosis inhibitor protein Bcl-XL were inhibited, cleaved-PARP and P53 were increased, and cytochrome C was released from the mitochondria into the cytoplasm. Moreover, under UV-A irradiation, I34 promoted the increase in ROS in Sf9 cells, activated the mitochondrial apoptotic signal transduction pathway, and finally, inhibited cell proliferation. Thus, novel furanocoumarins exhibit a potential application prospect as a biochemical pesticide.
呋喃香豆素是植物的次生代谢产物，被认为是天然的杀虫剂和杀菌剂，因为它们可以防止植物病原微生物的入侵和草食性昆虫的捕食。在本研究中，通过缩合、酯化、溴化和 Wittig 反应设计合成了新型 2-芳基呋喃香豆素衍生物。结果表明，2-噻吩基呋喃香豆素（I34）具有优异的光敏活性。细胞计数试剂盒-8 检测到 I34 在紫外 A（UV-A）光下 3 分钟内以时间和浓度依赖的方式抑制斜纹夜蛾（Sf9）细胞的增殖。倒置显微镜显示，用 I34 处理的细胞肿胀，膜破裂，出现凋亡小体。流式细胞术检测到 I34 可诱导 Sf9 细胞凋亡，增加细胞内活性氧（ROS）水平，降低线粒体膜电位，并阻断细胞周期停滞在 G2/M 期。透射电子显微镜检测到细胞线粒体嵴损伤、基质降解和线粒体空泡化。进一步的酶活性检测显示，凋亡相关蛋白 caspase-3 和 caspase-9 的酶活性显著增加（p<0.05）。最后，Western 印迹分析检测到 Akt 和 Bad 的磷酸化水平以及凋亡抑制蛋白 Bcl-XL 的表达受到抑制，裂解-PARP 和 P53 增加，细胞色素 C 从线粒体释放到细胞质中。此外，在 UV-A 照射下，I34 促进 Sf9 细胞中 ROS 的增加，激活线粒体凋亡信号转导途径，最终抑制细胞增殖。因此，新型呋喃香豆素作为生化农药具有潜在的应用前景。
