基于纳米材料拉曼增强特性的细胞氧化损伤检测

Cellular oxidative damage detection based on Raman-enhanced property of nanomaterials

表面增强拉曼可作为单细胞氧化损伤的一种检测手段。以铜离子诱导的细胞氧化损伤为模型、以表面增强拉曼光谱为检测手段,建立了用以表征细胞损伤的拉曼光谱方法。通过制备经多肽修饰的金纳米花作为拉曼增强基底,使其能够进入细胞并达到信号增强的效果。利用CCK-8细胞毒性实验测试基底对细胞的损伤程度,确定基底质量浓度为160μg/mL。进一步利用CCK-8细胞毒性实验验证Cu^(2+)对细胞的氧化损伤,证实经Cu^(2+)诱导后细胞出现明显的损伤和死亡现象。最后,建立了Cu^(2+)诱导的细胞氧化损伤模型,并获得不同条件下细胞所呈现的拉曼光谱,发现经氧化处理后,细胞受到不同程度的损伤,细胞内的糖类、核酸、脂质和蛋白质等物质均发生了一定程度的改变,对机体造成损伤。

Surface-enhanced Raman spectroscopy(SERS)can be used as an assay for single-cell oxidative damage.A Raman spectroscopic characterization method was developed to study cellular damage using copper ion-induced oxidative damage in cells as a model and surface-enhanced Raman spectroscopy as a detection method.Gold nanoflowers with peptide modifications were prepared as Raman-enhanced substrate to enable them to enter cells and achieve signal enhancement.The extent of cell damage caused by the substrate was tested using CCK-8 cytotoxicity assay,and concentration of the substrate was determined to be 160μg/mL.The cell oxidative damage caused by Cu^(2+)was also verified through CCK-8 cytotoxicity assay,and significant damage and death of cells were observed under the inducement of Cu^(2+).Finally,a model of Cu^(2+)-induced cell oxidative damage was established and Raman spectra presented by cells under different conditions were obtained.It was found that cells treated with oxidation were damaged to different degrees,and intracellular glycans,nucleic acids,lipids and proteins were altered to some extent,causing damage to the organism.