Investigating the autophagy pathway in silver@gold core-shell nanoparticles- treated cells using surface-enhanced Raman scattering
Investigating the autophagy pathway in silver@gold core-shell nanoparticles- treated cells using surface-enhanced Raman scattering
复制标题
使用表面增强拉曼散射研究银@金核壳纳米颗粒处理的细胞中的自噬途径
作者：
Yuqing Hou;Wen Zhang;Shengtao Li;Zhen Wang;Huiqing Zhong;Zhiming Liu;Zhouyi Guo
以往的研究表明纳米粒子可以诱导自噬，研究纳米粒子诱导自噬的主要方法是通过TEM和生化检测等传统方法。这些方法测量存在实验过程复杂、细胞破坏以及缺乏自噬途径各个阶段特征的缺点。表面增强拉曼散射（SERS）已广泛应用于生物应用中。前期研究通过结合SERS和化学计量学方法，如主成分分析-线性判别分析（PCA-LDA），通过从复杂的SERS数据集中分离光谱，实现了Au纳米粒子内吞作用中内体和溶酶体的识别和分布图谱。本研究通过还原银纳米粒子表面的金离子合成了银@金核壳纳米粒子（Ag@Au NPs），并以Ag@Au NPs作为自噬诱导剂和高性能SERS底物研究了Ag@Au NPs诱导的自噬。通过蛋白质印迹分析、流式细胞术和荧光染色证明了 Ag@Au NPs 诱导的促存活自噬。此外，Ag@Au NPs 处理的细胞中的自噬途径首先通过 SERS 结合改进的基于参考的 PCA-LDA 方法来阐明。本研究为利用SERS阐明纳米粒子诱导的自噬途径提供了一种可行的方法。
Previous studies have shown that nanoparticles can induce autophagy, and the main approach for investigating autophagy induced by nanoparticles is via traditional methods such as TEM and biochemical assay. These methods measurements suffer from the disadvantages of complicated experimental processes, cell destruction, as well as lack of characterization of individual stages of the autophagy pathway. Surface-enhanced Raman scattering (SERS) has been extensively used in biological applications. With the combination of SERS and chemometric methods, such as principal component analysis-linear discriminant analysis (PCA-LDA), identification and distribution mapping of endosomes and lysosomes in the endocytosis of Au nanoparticles has been achieved by segregating the spectra from complex SERS data sets in the previous study. In this study, silver@gold core–shell nanoparticles (Ag@Au NPs) were synthesized by reduction of gold ions on the surface of the silver nanoparticles, and the autophagy induced by Ag@Au NPs was studied with Ag@Au NPs serving both as an autophagy inducer and as a high-performance SERS substrate. Pro-survival autophagy induced by Ag@Au NPs was proved by the western blot assay, flow cytometry and fluorescent staining. Furthermore, the autophagy pathway in Ag@Au NPs-treated cells was first elucidated by SERS combined with a modified reference-based PCA-LDA methodology. This study provides a feasible way of using SERS to elucidate the autophagy pathway induced by nanoparticles.