目的：探讨四氧化三铁（Fe3O4）纳米粒子（PION）作为药物载体增强二氢卟吩e6（chlorin e6，Ce6）在胶质瘤中的增效作 用。方法：采用高温降解法和相转移法制备PEG-Fe3O4@Ce6复合纳米粒子（PION@E6），用水合粒径分析、透射电镜、胶体稳定 性分析、紫外可见光吸收光谱等方法对PION@E6进行鉴定。CCK-8法检测胶质瘤U251细胞的增殖活性，流式细胞术检测细胞 的凋亡水平，DCFH-DA探针法检测细胞中活性氧（reactive oxygen species，ROS）的水平。构建BALB/c-nu裸鼠胶质瘤U251细胞 移植瘤模型，动物活体荧光成像术及磁共振成像（MRI）观察PION@E6及Ce6在移植瘤中的潴留时间，比较PION@E6声动力治 疗组及Ce6声动力治疗组的第28天生存情况及肿瘤体积。结果：PION@E6的核心粒径为10 nm、水合粒径为（37.86±12.90）nm， 具有良好的水溶性和稳定性；吸收光谱及XRD图谱显示Ce6已经负载到Fe3O4纳米粒子上。与Ce6声动力组比较，PION@E6声 动力组U251细胞的增殖活性显著下降（P<0.05），细胞凋亡率显著升高（均P<0.05），细胞中ROS水平显著升高（P<0.05）。荷瘤裸 鼠胶质瘤U251细胞移植瘤治疗实验结果显示，与Ce6声动力治疗组比较，PION@E6声动力治疗组裸鼠移植瘤组织中潴留时间 显著延长（P<0.05），存活的裸鼠数显著增多，移植瘤体积显著缩小（P<0.01）。结论：Fe3O4纳米粒子对Ce6介导的胶质瘤U251细 胞声动力治疗具有明显的增效作用。
Objective: To explore the synergistic effect of Fe3O4 nanoparticles (PION), as a drug carrier, on enhancing the effect of chlorin e6 (Ce6) in glioma. Methods: PEG-Fe3O4@Ce6 composite nanoparticles (PION@E6) were prepared by high temperature degradation method and phase transfer method, and then verified by hydrated particle size analysis, transmission electron microscopy, colloidal stability analysis and ultraviolet-visible light absorption spectroscopy, etc. CCK-8 method was used to detect the proliferation activity of glioma U251 cells, Flow cytometry was used to detect the apoptosis of the cells, and the DCFH-DA probe method was used to detect the level of reactive oxygen species (ROS) in the cells. Glioma U251 cell transplanted tumor model was constructed on BALB/c-nu nude mice. The retention time of PION@E6 and Ce6 in transplanted tumors was observed with animal fluorescence imaging technology and magnetic resonance imaging (MRI), and the survival and tumor volume on the 28th day in the PION@E6 sonodynamic therapy group and Ce6 sonodynamic therapy group were compared. Results: Transmission electron microscopy and hydrated particle size analysis showed that the core particle size of PION@E6 was 10 nm and the hydrated particle size was (37.86±12.90) nm; colloidal stability analysis showed that PION@E6 had good water solubility and stability; and the absorption spectrum and XRD atlas showed that Ce6 had been loaded on Fe3O4 nanoparticles. Compared with the Ce6 sonodynamic group, the proliferation activity of U251 cells in the PION@E6 sonodynamic group was significantly decreased (P<0.05)， the apoptosis rate was significantly increased (all P<0.05)，and the level of ROS in the cells was significantly increased (P<0.05). In vivo experiments on nude mice bearing glioma U251 cell transplanted tumors showed that compared with Ce6 sonodynamic therapy group, the retention time of Ce6 in the transplanted tumor tissues of the PION@E6 sonodynamic therapy group was significantly prolonged (P<0.05)，the number of survived nude mice was significant increased, and the transplanted tumor volume was significantly reduced (P<0.01). Conclusion: Fe3O4 nanoparticles have a significant synergistic effect on Ce6-mediated sonodynamic therapy of glioma U251 cells.
国家自然科学基金资助项目（No.32060228）；广西自然科学基金资助项目（No. 2017GXNSFAA198112,No. 2019GXNSFAA245077）； 广西研究生教育创新计划资助项目（No. YCSW2019214, No.YCSW2020225）；广西脑与认知神经科学重点实验室开放课题资助项目（No. GKLBCN-20200108-02）