基于扫描电化学显微镜技术评价镉诱导MCF-10A细胞产生的过氧化氢水平  

Hydrogen peroxide levels in cadmium-induced MCF-10A cells were evaluated by scanning electrochemical microscopy

作　　者：张标 樊孝银 鲁理平[1] ZHANG Biao;FAN Xiaoyin;LU Liping(Regional Air Composite Pollution Prevention and Control Key Laboratories in Beijing,Department of Environmental Sciences,Beijing University of Technology,Beijing,100124,China)

机构地区：[1]区域大气复合污染防治北京市重点实验室环境科学系北京工业大学,北京100124

出　　处：《环境化学》2022年第5期1693-1701,共9页Environmental Chemistry

基　　金：国家自然科学基金(21876005,21936001);北京市卓越青年计划(BJJWZYJH 01201910005017)资助。

摘　　要：本研究利用扫描电化学显微镜(SECM)技术在生理条件下对人正常乳腺上皮细胞(MCF-10A)受氯化镉刺激的形态变化和活性氧物种(ROS)释放情况进行了实时检测.结果表明,MCF-10A细胞在低浓度或短时间的氯化镉刺激下呈现收缩防御模式;但受到高浓度或长时间的氯化镉刺激时,细胞对氯化镉毒性的防御能力降低,细胞内氧化还原平衡态受到破坏,造成严重的氧化损伤.并利用荧光探针DCFH-DA法证实了扫描电化学显微镜技术检测结果的正确性和可靠性.此外,通过对镉作用下细胞内还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶,超氧化物歧化酶(SOD),过氧化氢酶(CAT)及谷胱甘肽(GST)活性的测定,进一步分析了金属镉诱导MCF-10A细胞的氧化应激机制.In this study,scanning electrochemical microscopy(SECM)technology was used to achieve real-time detection of the morphological changes and the release of reactive oxygen species(ROS)of human normal breast epithelial cells(MCF-10A)stimulated by cadmium chloride under physiological conditions.The results showed that MCF-10A cells exhibited a contraction defense mode under the condition of a low concentration cadmium chloride stimulation or a short time stimulation;However,when dealt with a high concentration cadmium chloride stimulation or the stimulation time was long,the defense ability of the cells against the toxicity of cadmium chloride decreased,the internal redox balance of the cells was destroyed,causing a serious oxidative damage.And the correctness and reliability of the detection results of scanning electrochemical microscopy technology was verified by using the fluorescent probe DCFH-DA method.In addition,the mechanism of oxidative stress induced by metal cadmium in MCF-10A cells was further analyzed through the determination of activity of the reduction of nicotinamide adenine dinucleotide phosphate(NADPH)oxidase,superoxide dismutase(SOD),catalase(CAT),glutathione(GST)in the cell under the action of cadmium.

关 键 词：扫描电化学显微镜(SECM) H2O_(2) 氧化损伤 

分 类 号：X50[环境科学与工程—环境工程]