基于Si纳米粒子拉曼成像的HepG2细胞凋亡检测及其活性分析  

Apoptosis Assay and Activity Analyses of HepG2 Cells Based on Raman Mapping of Si Nanoparticles

作　　者：林凡佳 吴冠毅 吴莎 张珠峰 石清华 周骏[2] LIN Fan-jia;WU Guan-yi;WU Sha;ZHANG Zhu-feng;SHI Qing-hua;ZHOU Jun(School of Marine Sciences,Ningbo University,Ningbo 315211,China;Institute of Photonics,Faculty of Science,Ningbo University,Ningbo 315211,China;Affiliated Hospital,School of Medicine,Ningbo University,Ningbo 315211,China)

机构地区：[1]宁波大学海洋学院,应用海洋生物技术教育部重点实验室,浙江宁波315211 [2]宁波大学理学院,光学与光电子技术研究所,浙江宁波315211 [3]宁波大学医学院附属医院,浙江宁波315211

出　　处：《光学与光电技术》2018年第4期37-45,共9页Optics & Optoelectronic Technology

基　　金：国家自然科学基金(61320106014、61675104);浙江省大学生新苗人才计划(2017R405047)资助项目

摘　　要：根据细胞的生物吞噬特性,通过研究人肝癌细胞(HepG2)内吞Si纳米粒子的拉曼成像特性,提出了一种新的细胞凋亡检测方法。首先,在Si纳米粒子与细胞共培养过程中,对HepG2细胞施加不同浓度的凋亡诱导剂——黄连素,然后对细胞内吞的Si纳米粒子在520cm^(-1)处的特征拉曼峰检测成像,通过分析细胞区域拉曼像图像素的平均信号强度,表征细胞内吞Si纳米粒子的能力,再对照流式细胞术的检测结果从而获得在不同浓度细胞凋亡诱导剂作用下的细胞凋亡率。结果表明,随着黄连素浓度的增加,细胞区域拉曼像图的平均信号强度逐渐减弱,说明在HepG2细胞凋亡过程中线粒体的活性不断减弱,能量代谢受阻,造成细胞对Si纳米粒子的内吞能力减弱。特别地,在高浓度(150μM)黄连素作用下,细胞区域内520cm^(-1)的特征拉曼峰十分微弱,说明此时HepG2细胞基本丧失内吞Si纳米粒子的能力。因此,基于细胞内吞Si纳米粒子拉曼成像方法,可以对不同环境下的体外细胞进行凋亡检测,将在细胞毒理研究和药效评价方面具有潜在应用。Based on the biological phagocytosis of cells,a new method of cell apoptosis detection is proposed by studying the characteristics of Raman mapping of the intracellular swallowed Si nanoparticles（NPs）in the human hepatoma cells（HepG2）.First,the different concentrations of apoptotic inducers（berberine）is applied to HepG2 cells during the process of culturing cells with Si NPs.Then the characteristic Raman peak at 520 cm^-1 of the intracellular swallowed Si NPs in the HepG2 cells are detected by the Raman mapping technology.Next,by analyzing the average pixel signal intensity of Raman image of endocytosed Si NPs in the cells,the apoptosis states of HepG2 cells dependent on the different concentrations of berberine are presented.And by means of flow cytometry,the relationship of the apoptosis rates of cells and the average pixel signal intensities of Raman image scorresponding to the different concentrations of berberine is obtained.The results show that,with the increasing of the berberine concentration,the average intensity of Raman images of cell decrease gradually,which demonstrates that the mitochondrial activity and the energy metabolism of HepG2 cells decline so that the endocytose ability of HepG2 cells for Si NPs descend.Especially,when HepG2 cells are induced by 150μm of berberine,the peak intensity of 520 cm^-1 of endocytosed Si NPs nearly disappeared,which presents that HepG2 cells almost lose the phagocytosis ability.Therefore,by the evaluation of Raman mapping of endocytosed Si NPs in cells,the apoptosis assay of cells in different environments could be detected,which has potential applications in the fields of the cell toxicology and pharmacology.

关 键 词：Si纳米粒子 特征拉曼峰 拉曼成像 细胞凋亡 细胞摄取 

分 类 号：O657.37[理学—分析化学] R329.2[理学—化学]