摘要
研究阿魏酸是否具有拮抗PM2.5致线粒体损伤作用及拮抗机制。实验分为空白对照组、PM2.5(240 μg/m L(损伤组和阿魏酸(80 mmol/L)保护组。通过Mito SOX Red特异性探针和流式细胞仪测定细胞线粒体内活性氧(reactive oxygen species,ROS)的含量;Annexin-V/PI双染色法流式细胞仪检测细胞凋亡率;JC-1染色法流式细胞仪检测细胞线粒体膜电位的变化;活体细胞线粒体膜通道孔荧光检测试剂盒检测细胞线粒体膜通道孔(mitochondrial permeablity transition pore,m PTP)的开放情况;Western blot法检测caspase-3、caspase-9、含锰金属辅基的超氧化物歧化酶(manganese superoxide dismutase,Mn SOD)和过氧化物酶增殖体激活受体γ共激活因子-1α(subunit of peroxisome proliferators-activated receptor-γ-coactivator-1α,PGC-1α)相关蛋白表达量。结果显示,与空白对照组比较,PM2.5损伤组的细胞ROS含量、caspase-3和caspase-9表达量及细胞凋亡率升高,线粒体膜电位和m PTP活性及PGC-1α、Mn SOD的表达量降低;阿魏酸预处理后,显著降低了PM2.5造成的A549细胞ROS的含量、细胞凋亡率及caspase-3和caspase-9表达量的升高,显著升高了PM2.5致A549细胞线粒体的膜电位、m PTP的活性及PGC-1α和Mn SOD表达量的降低。阿魏酸对PM2.5致A549细胞线粒体损伤具有明显的保护作用。
In this study, we examined whether ferulic acid (FA) could protect against PM2.5-induced mitochondrial damage in A549 cells and explored the underlying mechanism. The cells were treated with PM2.5 (240 μg/mL) with and without the addition of 80 mmol/L FA. The contents of reactive oxygen species (ROS) were determined by flow cytometry using MitoSOX Red specific probe. Apoptosis was detected by flow cytometry with Annexin-V/PI method. Mitochondrial membrane potential was detected by flow cytometry with JC-1 staining. The activity of mitochondrial membrane channel pore was tested by mitochondrial permeability transition pore (mPTP) kit. The expression of caspase-3, caspase-9, manganese superoxide dismutase (MnSOD), and peroxisome proliferators-activated receptor-γ-coactivator-1α (FGC-1α) were detected by Western blotting. The results showed that, compared with the blank control group, the level of ROS and the expression of caspase-3 and caspase-9 in mitochondria, as well as the apoptosis rate of A549 cells induced by PM2.5 were increased. The membrane potential and the activity of mPTP in mitochondria of A549 cells induced by PM2.5 were decreased as well as the expression of PGC-1α and MnSOD. The pre-treatment of FA could prevent the increased mitochondrial ROS, apoptosis rate and expression of caspase-3 and caspase-9, and the decreased mitochondrial membrane potential, mPTP activity, and expression of PGC-1α and MnSOD caused by PM2.5. Therefore, FA had a significant protective effect on PM2.5-induced mitochondrial damage in A549 cells.
作者
劳文艳
毕婷婷
周艳丽
陈世杰
赵晓红
刁屹
LAO Wenyan;BI Tingting;ZHOU Yanli;CHEN Shijie;ZHAO Xiaohong;DIAO Yi(Research Institute for Science and Technology of Functional Foods, Beijing Union University, Beijing 100191, China;Key Laboratory of Bioactive Substances and Functional Foods, Beijing Union University, Beijing 100191, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2017年第3期195-200,共6页
Food Science
基金
中国营养学会营养科研基金-帝斯曼专项科研基金项目(2015-030(CNS-DSM))
北京联合大学生物活性物质与功能食品北京市重点实验室开放课题(Zk70201501)