摘要
以嗜热四膜虫为模式生物从氧化应激、凋亡及线粒体损伤作用探讨姜黄素的毒性效应,为姜黄素用药及生态安全提供依据.急性毒性实验:姜黄素(0.625、1.25、2.5μg/mL)作用四膜虫2 h,通过快速细胞分析仪计数细胞,光学显微镜观察细胞形态变化;氧化应激:荧光酶标仪检测细胞内活性氧(ROS)水平及乳酸脱氢酶活性(LDH);线粒体损伤:JC-1探针法检测线粒体膜电位(MMP)的变化;凋亡实验:Annexin V-FITC/碘化丙啶(PI)双染流式细胞仪检测细胞凋亡率.实验结果显示,姜黄素对四膜虫的毒性呈剂量依赖效应,0.625、1.25、2.5μg/mL分别抑制了16%、32%、37%,2.5μg/mL处理组细胞形态发生显著变化;此外1.25、2.5μg/mL处理组的细胞内ROS分别增加了35%、44%(P<0.05).LDH的结果表明各浓度的姜黄素均能破坏细胞膜通透性,与对照组相比分别增加了10%、17%、45%.2.5μg/mL的姜黄素处理组能够引起线粒体膜电位(MMP)去极化(P<0.05),凋亡率增加7.42%(P<0.05).姜黄素对嗜热四膜虫存在毒性作用,抑制细胞增殖,并通过氧化应激作用破坏细胞膜,增加细胞膜通透性,2.5μg/mL的姜黄素能够诱导四膜虫细胞凋亡并伴随线粒体损伤,暗示姜黄素可能通过激活线粒体途径诱导嗜热四膜细胞的凋亡.
Objective:To explore the toxic effects of curcumin from oxidative stress,apoptosis and mitochondrial damage in Tetrahymena Thermophila.Methods:acute toxicity test,curcumin(0.625,1.25,2.5μg/mL)was used to treat Tetrahymena Thermophila for 2 hours.The cells were counted by rapid cell analyzer and the morphological changes were observed by light microscope;oxidative stress,intracellular reactive oxygen species(ROS)level and lactate dehydrogenase activity(LDH)were detected by fluorescence enzyme labeling instrument;mitochondrial membrane potential(mm)was detected by JC-1 probe method.The apoptosis rate was detected by flow cytometry with annexin V-FITC/PI double staining.Results:the toxicity of curcumin on Tetrahymena Thermophila was dose-dependent.The toxicity of curcumin was inhibited by 16%,32%and 37%for 0.625,1.25 and 2.5μg/mL,respectively.In addition,ROS in 1.25 and 2.5μg/mL groups increased by 35%and 44%,respectively.The results of LDH showed that curcumin could destroy the permeability of the cell membrane,which increased by 10%,17%and 45%respectively compared with the control group.2.5μg/mL curcumin could induce mitochondrial membrane potential(MMP)depolarization(P<0.05),and the apoptosis rate increased by 7.42%(P<0.05).Conclusion:curcumin has toxic effect on Tetrahymena Thermophila,inhibits cell proliferation,destroys cell membrane and increases membrane permeability through oxidative stress.Curcumin at 2.5μg/mL can induce apoptosis of Tetrahymena Thermophila cells with mitochondrial damage,suggesting that curcumin may induce apoptosis of Tetrahymena Thermophila cells by activating mitochondria.
作者
葸灵娟
张梦玘
林志康
赵宇侠
XI Lingjuan;ZHANG Mengqi;LIN Zhikang;ZHAO Yuxia(Graduate School,Shanghai University of Traditional Chinese Medicine,Shanghai 200120;School of Pharmacy,Shanghai University of Medicine&Health Sciences,Shanghai 201138)
出处
《四川师范大学学报(自然科学版)》
CAS
2022年第3期394-401,共8页
Journal of Sichuan Normal University(Natural Science)
基金
国家自然科学基金(20130371)。
关键词
嗜热四膜虫
姜黄素
细胞凋亡
线粒体
活性氧
Tetrahymena Thermophila
curcumin
cell apoptosis,mitochondrial
reactive oxygen species