Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, Sichuan 610065, China.
Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, Sichuan 610065, China.
Pestic Biochem Physiol. 2019 Jun;157:169-177. doi: 10.1016/j.pestbp.2019.03.025. Epub 2019 Apr 1.
To explore the toxicity mechanisms of neochamaejasmin A (NCA), extracted from Stellera chamaejasme L., we first evaluated its cytotoxicity on the Spodoptera frugiperda (Sf9) cell line. The results confirmed that NCA inhibited Sf9 cell survival in both a dose- and time-dependent manner. Then, intracellular biochemical assays showed that NCA induced apoptosis in Sf9 cells. Evidence of apoptosis was confirmed by morphological changes and the activation of caspases-3/9. We also observed that NCA induced apoptosis via mitochondrial-dependent intrinsic apoptotic pathway by upregulating cytochrome c and proapoptotic protein (Bax) and downregulating the mitochondrial membrane potential (MMP) and antiapoptotic protein (Bcl-2). Moreover, we found a dose-dependent increase in reactive oxygen species (ROS), accumulation of lipid peroxidation product and an inactivation of the antioxidant enzymes in treated cells. Additionally, the cleavage of PARP and G2/M arrest were also detected in Sf9 cells exposed to NCA. These findings provide critical information that NCA effectively induced apoptosis in Sf9 cells through mitochondrial pathways.
为了探究从瑞香狼毒（ Stellera chamaejasme L.）中提取的新狼毒素 A（NCA）的毒性机制，我们首先评估了其对草地贪夜蛾（Spodoptera frugiperda， Sf9）细胞系的细胞毒性。结果证实，NCA 以剂量和时间依赖的方式抑制 Sf9 细胞存活。然后，细胞内生化测定表明，NCA 诱导 Sf9 细胞凋亡。形态学变化和 caspase-3/9 的激活证实了细胞凋亡的发生。我们还观察到，NCA 通过上调细胞色素 c 和促凋亡蛋白（Bax）以及下调线粒体膜电位（MMP）和抗凋亡蛋白（Bcl-2），诱导 Sf9 细胞通过线粒体依赖性内在凋亡途径发生凋亡。此外，我们发现，处理细胞中活性氧（ROS）的浓度呈剂量依赖性增加，脂质过氧化产物积累，抗氧化酶失活。此外，在暴露于 NCA 的 Sf9 细胞中还检测到 PARP 的裂解和 G2/M 期阻滞。这些发现为 NCA 通过线粒体途径有效诱导 Sf9 细胞凋亡提供了重要信息。
