棒曲霉毒素对人胚肾细胞增殖与诱导其凋亡的研究  

Effects of Patulin on the Proliferation and Apoptosis of Human Embryonic Kidney Cells

作　　者：张百刚[1,2] 彭晓丽[1] 李光辉[1] 徐云凤[1] 夏效东[1] 

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100 [2]兰州理工大学生命科学与工程学院,甘肃兰州730050

出　　处：《现代食品科技》2015年第3期12-17,共6页Modern Food Science and Technology

基　　金：国家自然基金(31201286);中央高校基本科研创新一般项目(2014YB021)

摘　　要：本文研究了棒曲霉素抑制人胚肾细胞增殖与诱导其凋亡的作用。不同浓度的棒曲霉素作用于人胚肾细胞293后,采用乳酸脱氢酶释放检测棒曲霉素对细胞增殖的抑制作用。通过扫描电镜和荧光显微镜观察了细胞形态学的变化,Annexin V-EGFP/PI标记检测细胞凋亡率,JC-1线粒体膜电位荧光探针检测了细胞线粒体膜电位的变化。实时荧光定量PCR测定了线粒体相关基因FIS1、ASL、SLC25A6、COX17的表达水平。结果表明,棒曲霉素可显著抑制人胚肾细胞293增殖并诱导凋亡,具有明显的量效关系,2.5、5、7.5、10和15μM棒曲霉素对人胚肾细胞293的抑制率分别为8.1%、18.2%、31.0%、42.2%和63.1%;5μM棒曲霉素处理0、3、10、24 h,细胞的存活率分别为94.2%、78.1%、47.4%。5μM和10μM的棒曲霉素处理8h后,细胞凋亡率分别为16.4%,和20.1%。线粒体膜电位和线粒体相关基因的表达水平的变化表明棒曲霉素可能通过线粒体途径引起人胚肾细胞293凋亡。To study the mechanism of patulin on the proliferation and apoptosis ofhuman embryonic kidney cells, HEK293 cells were cultured and treated with different concentrations of patulin in vitro. The effect of patulin on cell proliferation level was examined by LDH cytotoxicity assay. The morphological changes in patulin-treated HEK293 cells were observed using scanning electron microscopy and fluorescence microscopy. The cell apoptosis rate was detected by AnnexinV-EGFP/PI double staining. The mitochondrial membrane potential was measured by a JC-1 probe. The expression levels of the mitochondrial related genes FIS1, ASL, SLC25A6, and COX17 were measured by real-time fluorescent quantitative PCR. The results showed that PAT inhibited the growth of HEK293 cells in a dose-dependent manner and resulted in apoptosis in HEK293 cells. Treatments with 2.5, 5, 7.5, 10, and 15μM PAT for 8h resulted in significant decreases in cell viability by 8.1%, 18.2%, 31.0%, 42.2%, and 63.1%, respectively. When treated with 5μM patulin for 0, 3, 10, and 24h, the cell survival rates were 94.2%, 78.1%, and 47.4%, respectively. After treatment with 5 and 10μM PAT for 8h, the number of apoptotic cells increased in a dose-dependent manner by 16.4% and 20.1%. The changes in the expression levels of mitochondrial membrane potential and mitochondrial genes indicate that patulin may cause HEK293 apoptosis through mitochondrial pathways.

关 键 词：棒曲霉素 细胞增殖 凋亡 人胚肾细胞 

分 类 号：R96[医药卫生—药理学]