基于介孔上转换材料的肿瘤微环境响应诊疗一体化系统的研究

发布时间：2018-03-13 15:12

本文选题：核壳介孔上转换　切入点：谷胱甘肽　出处：《郑州大学》2017年硕士论文　论文类型：学位论文

【摘要】：研究目的:制备肿瘤微环境响应诊疗一体化系统(mUCNPs@DOX/CuS/HA),对其理化性质、体内外抗肿瘤活性、药物代谢动力学及体内成像进行考察,研究该载药系统在UCL/MRI/PAT三模态成像介导下的抗肿瘤效果。研究方法:(1)本研究采用高温热分解法制备核壳介孔上转换纳米粒(mUCNPs),通过荧光光谱、X射线光电子能谱分析(XPS)、X射线衍射分析(XRD)、动态光散射技术(DLS)、透射电子显微镜(TEM)、氮气吸附脱附分析对mUCNPs进行表征;在mUCNPs表面修饰二硫键,连接CuS与透明质酸,制备得到mUCNPs@DOX/CuS/HA,通过紫外光谱、荧光光谱、红外光谱、动态光散射技术(DLS)、透射电子显微镜(TEM)对纳米复合物进行表征,并考察其光热性能及热稳定性;采用HPLC法测定其载药量和药物释放。(2)体外抗肿瘤活性及靶向性研究:采用SRB法考察诊疗系统对MCF-7细胞活性的影响;使用激光共聚焦显微镜及流式细胞仪考察细胞对诊疗系统摄取情况;此外,还考察了诊疗系统对细胞周期及凋亡的影响。(3)药物代谢动力学研究:以SD大鼠为动物模型,进行药物代谢动力学实验,采用HPLC法测定大鼠血浆药物浓度,并计算相应的药物代谢动力学参数。(4)药效学及体内成像研究:采用S180荷瘤小鼠为动物模型,通过肿瘤相对体积变化及HE染色考察该诊疗系统的毒性及肿瘤抑制作用;考察诊疗系统在体内的成像效果及靶向性。研究结果:NaGdF4介孔层可以大幅提高上转换发光强度,氮气吸附脱附实验证明了mUCNPs表面介孔分布及载药能力。紫外光谱、红外光谱、热重分析表明mUCNPs表面成功修饰了CuS和HA,透射电镜观察到诊疗系统平均粒径约60 nm,该系统具有良好的光热性能及热稳定性;此外,该诊疗系统具有体外核磁成像能力及良好的稳定性。mUCNPs@DOX/CuS/HA在10 mM GSH,pH 5.0PBS中释放最多,光热作用可进一步促进药物释放,说明该诊疗系统能够响应肿瘤细胞微环境释放药物。细胞毒实验结果证明mUCNPs@CuS/HA具有良好的生物相容性;当化疗与热疗联合,mUCNPs@DOX/CuS/HA在980 nm激光照射下,显示出强大的细胞杀伤作用;细胞摄取实验结果显示,HA修饰的mUCNPs@DOX/CuS/HA靶向性最强;细胞周期与细胞凋亡实验进一步证明了化疗与热疗协同治疗效果。药物代谢动力学结果表明,诊疗系统延长了药物在大鼠体内停留的时间,使药物的清除速率减慢。mUCNPs@DOX/CuS/HA在980nm激光照射下,协同化疗和热疗,显示出强大的肿瘤抑制作用;小鼠体重变化及HE染色结果显示诊疗系统对正常组织器官几乎无毒副作用;体内成像结果显示,mUCNPs@DOX/CuS/HA在进入小鼠体内2 h后在肿瘤部位累积量达到最大,显示出较强的成像能力。结论:本研究构建的mUCNPs@DOX/CuS/HA诊疗系统,具有肿瘤靶向和氧化还原刺激响应型释药特点,可用于UCL/MRI/PAT三模态成像介导的化疗和光热治疗,实时评估肿瘤治疗效果。
[Abstract]:Objective: to prepare an integrated system for the diagnosis and treatment of tumor microenvironment response, and investigate its physicochemical properties, anti-tumor activity in vivo and in vitro, pharmacokinetics and imaging in vivo. The antitumor effect of the drug carrier system mediated by UCL/MRI/PAT three-mode imaging was studied. (methods: 1) in this study, the core-shell mesoporous upconversion nanocrystalline (mUCNPsN) was prepared by high temperature thermal decomposition. X-ray photoelectron spectroscopy (XPS) was used to analyze the results of X-ray photoelectron spectroscopy (XPS). MUCNPs was characterized by X-ray diffraction analysis, dynamic light scattering technique, transmission electron microscope (TEM) and nitrogen adsorption desorption analysis. Modified disulfide bond on the surface of mUCNPs, connecting CuS with hyaluronic acid, the nanocomposites were characterized by UV spectra, fluorescence spectra, infrared spectra, dynamic light scattering techniques, transmission electron microscopy (TEM). The photothermal properties and thermal stability were investigated, and the antitumor activity and targeting of MCF-7 cells were determined by HPLC method. The effects of diagnosis and treatment system on the activity of MCF-7 cells were investigated by SRB method. Laser confocal microscopy and flow cytometry were used to investigate the uptake of cells to the diagnosis and treatment system. In addition, the effects of the diagnosis and treatment system on cell cycle and apoptosis were investigated. The pharmacokinetic experiment was carried out. The plasma drug concentration in rats was determined by HPLC method. The pharmacodynamics and in vivo imaging of the corresponding pharmacokinetic parameters were calculated: S180 tumor-bearing mice were used as animal models. The toxicity and tumor inhibition of the diagnosis and treatment system were investigated by tumor relative volume change and HE staining. The imaging effect and targeting of the diagnosis and treatment system in vivo were investigated. The results showed that the mesoporous layer: NaGdF4 could significantly increase the intensity of up-conversion luminescence. Nitrogen adsorption and desorption experiments have proved the mesoporous distribution and drug loading ability of mUCNPs surface. Thermogravimetric analysis showed that the surface of mUCNPs was successfully modified with CuS and HA.The average particle size of the diagnostic and treatment system was about 60 nm, and the system had good photothermal properties and thermal stability. The diagnostic and therapeutic system has in vitro nuclear magnetic imaging capability and good stability. The dox / CuSr / HA released the most in pH 5.0 PBS of 10 mm GSHN. Photothermal action can further promote drug release. The results of cytotoxicity test showed that mUCNPs@CuS/HA had good biocompatibility, and that when chemotherapeutic therapy combined with hyperthermia combined with DOX / Cu / Cu / HA, it showed strong cytotoxicity under the irradiation of 980nm laser. The results of cell uptake test showed that HA-modified mUCNPs@DOX/CuS/HA was the most targeted, cell cycle and apoptosis experiments further proved the synergistic effect of chemotherapy and hyperthermia. The diagnosis and treatment system prolonged the stay time of the drug in rats and slowed down the drug clearance rate. Under the irradiation of 980nm laser irradiation, the drug clearance rate of .mUCNPs@ DOX / CuS- / HA combined with chemotherapy and hyperthermia showed strong tumor inhibition. The results of body weight change and HE staining showed that the diagnosis and treatment system had almost no toxic side effects on normal tissues and organs, and the imaging results in vivo showed that the accumulation of mUCNPsR DOX / CuS- / HA reached the maximum in the tumor site 2 hours after entering the body of mice. Conclusion: the mUCNPs@DOX/CuS/HA system constructed in this study has the characteristics of tumor targeting and redox stimulation-responsive drug release and can be used in UCL/MRI/PAT three-mode imaging mediated chemotherapy and photothermal therapy. To evaluate the effect of tumor therapy in real time.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943;R96

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