氧化铱纳米载药复合物的制备及其用于肿瘤声动力-化疗协同治疗的研究  

Preparation of drug-loaded iridium oxidenanocomposites and its application in synergetic sonodynamic/chemotherapeutic therapy against cancer

作　　者：邹卫娟 郝俊年 吴建荣 郑元义[1,2] ZOU Weijuan;HAO Junnian;WU Jianrong;ZHENG Yuanyi(Chongqing Key Laboratory of Ultrasound Molecular Imaging,Chongqing Medical University,Chongqing,400010;Shanghai Institute of Ultrasound Medicine,Sixth People’s Hospital affiliated to Shanghai Jiaotong University,Shanghai,200233,China)

机构地区：[1]重庆医科大学超声分子影像重庆市重点实验室,重庆400010 [2]上海交通大学附属第六人民医院,上海超声医学研究所,上海200233

出　　处：《第三军医大学学报》2021年第22期2395-2406,共12页Journal of Third Military Medical University

基　　金：国家自然科学基金委重点国际(地区)合作研究项目(81720108023);国家自然科学基金重点项目(82030050);癌基因及相关基因国家重点实验室开放项目(KF2104-93#)。

摘　　要：目的制备氧化铱(iridium oxide,IrO_(X))作为纳米声敏剂,用以负载化疗药物阿霉素(doxorubicin,DOX)后得到具有pH和超声辐照双响应的纳米载药复合物(IrO_(X)@DOX),验证其用于肿瘤声动力治疗(sonodynamic therapy,SDT)-化疗协同治疗肿瘤的效果。方法采用热水解法制备IrO_(X)纳米颗粒,并负载DOX。采用透射电子显微镜(transmission electron microscope,TEM)、X射线光电子能谱(X-ray photoelectron spectroscopy,XPS)、紫外-可见分光光度计(ultraviolet-visible spectrophotometer,UVvis)、Zeta电位分析仪等手段对纳米颗粒的形貌及理化性质进行相关表征;采用UV-Vis和酶标仪考察载药量及药物释放行为;用电子顺磁共振波谱仪(lectron spin-resonance spectroscopy,ESR)定性评价其活性氧产生性能;激光共聚焦显微镜评价IrO_(X)@DOX在超声辐照下的细胞摄取行为;荧光显微镜和流式细胞仪探究细胞内活性氧产生;CCK-8法及流式凋亡检测评价IrO_(X)@DOX的声动力治疗-化疗协同治疗效果。建立Hepa1-6肝癌裸鼠皮下移植瘤模型,并分为生理盐水组、IrO_(X)组、IrO_(X)+US组、IrO_(X)@DOX组、IrO_(X)@DOX+US组(n=3),观察各组荷瘤鼠肿瘤的体积、超声造影成像结果与病理学变化,计算肿瘤体积抑制率评价其体内协同治疗效果。结果成功制备IrO_(X)纳米颗粒,粒径大约为5 nm。当IrO_(X)与DOX的比值为4.0时,载药量可达18.7%;IrO_(X)@DOX在p H=7.4时释放的DOX量不足10%,证实了其良好的稳定性;pH=5.5时,DOX的累积释放量可达39.9%。在超声辐照下释放量进一步增加达69.9%;ESR和UV-Vis实验结果表明活性氧产量与辐照功率和时间呈正相关;激光共聚焦结果显示超声能促进细胞对IrO_(X)@DOX的摄取;细胞毒性实验表明,与单纯化疗组相比,协同治疗组存活率降至19.4%,证明IrO_(X)@DOX在超声辐照下可以实现良好的SDT-化疗协同治疗效果。动物实验表明与生理盐水组相比,IrO_(X)+US组、IrO_(X)@DOX组和IrO_(X)@Objective To prepare iridium oxide(IrO_(X)) as a nano-sonosensitizer to load chemotherapy drug doxorubicin(DOX) in order to obtain nanocomposites IrO_(X)@ DOX with dual response of pH/ultrasound(US) irradiation,and to verify its efficacy in sonodynamic therapy(SDT)-chemotherapy for tumor treatment.Methods IrO_(X)nanoparticles were prepared by thermal hydrolysis and then loaded with DOX.The morphological, chemical and physical properties of the nanoparticles were characterized by transmission electron microscopy(TEM), X-ray photoelectron spectroscopy(XPS), ultraviolet-visible spectrophotometer(UV-Vis) and Zeta potential analyzer.The drug loading efficiency and drug release behavior were determined with UV-vis spectrometry and ELISA.The generation of reactive oxygen species(ROS) by IrO_(X)@ DOX nanoparticles was evaluated usingelectron spin-resonance spectroscopy(ESR).The uptake of IrO_(X)@ DOX by 4 T1 cells under ultrasonic irradiation was evaluated with laser confocal scanning microscopy(LCSM),and the level of intracellular ROS was investigated using fluorescence microscopy and flow cytometry.The synergistic efficacy of IrO_(X)@ DOX in SDT-chemotherapy against 4 T1 cells was evaluated by CCK-8 assay and flow cytometry.In addition,the Hepa1-6 xenograft tumor model in nude mice was built and divided into 5 groups(n = 3 in each group) as follows: saline,IrO_(X),IrO_(X)+US,IrO_(X)@ DOX,and IrO_(X)@DOX+US.The tumor volume,contrast-enhanced US imaging and pathological changes of each group were observed,and the inhibitory rate of tumor volume was calculated to evaluate the effect of synergistic therapy in vivo.Results IrO_(X)nanoparticles were successfully prepared,at a diameter of about 5.0 nm,and when the ratio of IrO_(X)to DOX was 4.0,the drug loading reached 18.7%.Prepared IrO_(X)@ DOX released less than 10%of DOX at pH 7.4,which confirmed its good stability,while the cumulative release of DOX reached 39.9% at pH 5.5,and further increased to 69.9% under US irradiation.Both ESR and UV-vis experiments demonstrated th

关 键 词：氧化铱 声动力治疗 化疗 协同治疗 肿瘤 

分 类 号：R454.3[医药卫生—治疗学] R730.58[医药卫生—临床医学]