Curcumin induces autophagic cell death in Spodoptera frugiperda cells
Curcumin induces autophagic cell death in Spodoptera frugiperda cells
复制标题
姜黄素诱导草地贪夜蛾细胞自噬细胞死亡
DOI：
10.1016/j.pestbp.2017.05.004
复制
发表时间：
2017-06-01
影响因子：
4.7
通讯作者：
Zhong Guohua
中科院分区：
文献类型：
--
作者：
Sethuraman, Veeran;Shu Benshui;Zhong Guohua
自噬（II型细胞死亡）在昆虫毒理学调控中的作用日益受到关注，推动了自噬细胞死亡途径的研究。姜黄是草本植物姜黄（Curcuma longa，印度泰米尔语为Manjal，中国为Jianghuang）的根茎，传统上单独或与其他植物杀虫剂组合用于害虫防治。然而，姜黄或姜黄素在害虫中发挥细胞毒性的机制尚不清楚。在这项研究中，我们调查姜黄（姜黄素）作为一种天然农药的效力使用Sf 9昆虫系。采用细胞增殖实验、倒置相差显微镜观察和透射电镜观察等方法，研究了姜黄素对草地贪夜蛾（Spodoptera frugiperda，Sf 9）细胞自噬的诱导作用。使用荧光染料单丹酰尸胺（MDC）评价自噬。使用浓度为5-15 μ g/mL的3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四唑溴化盐（MTT）检测细胞死亡测量。姜黄素抑制Sf 9细胞的生长，并诱导自噬性细胞死亡，呈时间和剂量依赖性。MDC染色显示自噬空泡的存在，并呈剂量和时间依赖性增加。在超微结构水平，透射电子显微镜下，细胞显示大量的自噬空泡积累和染色质凝聚的情况下。ATG 8-I和ATG 8-II的蛋白质表达水平，自噬相关蛋白的成熟标志物，姜黄素治疗后以时间依赖性方式升高。本研究证明姜黄素诱导昆虫细胞系Sf 9的自噬性细胞死亡，这是姜黄素在昆虫细胞中的细胞毒作用的首次报道，将作为未来的天然杀虫剂。(C)2017由Elsevier Inc.出版
The increasing interest in the role of autophagy (type II cell death) in the regulation of insect toxicology has propelled study of investigating autophagic cell death pathways. Turmeric, the rhizome of the herb Curcuma longa (Manjal in Tamil, India and Jianghuang in Chinese) have been traditionally used for the pest control either alone or combination with other botanical pesticides. However, the mechanisms by which Curcuma longa or curcumin exerts cytotoxicity in pests are not well understood. In this study, we investigated the potency of Curcuma longa (curcumin) as a natural pesticide employing Sf9 insect line. Autophagy induction effect of curcumin on Spodoptera frugiperda (Sf9) cells was investigated using various techniques including cell proliferation assay, morphology analysis with inverted phase contrast microscope and Transmission Electron Microscope (TEM) analysis. Autophagy was evaluated using the fluorescent dye monodansylcadaverine (MDC). Cell death measurement was examined using 3-(4,5-Dimethylthiazol-2-1)-2,5-Diphenyltetrazolium Bromide (MTT) within the concentrations of 5-15 mu g/mL. Curcumin inhibited the growth of the Sf9 cells and induced autophagic cell death in a time and dose dependent manner. Staining the cells with MDC showed the presence of autophagic vacuoles while increased in a dose and time dependent manner. At the ultrastructural level transmission electron microscopy, cells revealed massive autophagy vacuole accumulation and absence of chromatin condensation. Protein expression levels of ATG8-I and ATG8-II, well-established markers of autophagy related protein were elevated in a time dependent manner after curcumin treatment. The present study proves that curcumin induces autophagic cell death in Sf9 insect cell line and this is the first report of cytotoxic effect of curcumin in insect cells and that will be utilized as natural pesticides in future. (C) 2017 Published by Elsevier Inc.