一种带新型修饰配体的金纳米杆的组织分布和急性毒性试验及其在红外热疗辅助下对鼻咽癌移植瘤抑制作用的体内研究

发布时间：2018-05-30 08:54

本文选题：多巯基磺酸甜菜碱 + 金纳米杆　；参考：《浙江大学》2014年硕士论文

【摘要】：目的：纳米颗粒的低毒性是其在生物利用的先决条件。我们试图开发一种带新型修饰配体即多巯基磺酸甜菜碱聚合物(简称PTSB)的金纳米杆,评价该PTSB-金纳米杆在小鼠体内的组织分布和急性毒性作用,进一步探讨该PTSB-金纳米杆在红外热疗辅助下对裸鼠鼻咽癌移植瘤的治疗作用,从而寻找治疗鼻咽癌的一种新方法。材料和方法：两性离子磺酸甜菜碱(sB)形成聚合物通过多个巯基与金纳米杆牢固结合,构建PTSB-金纳米杆。以ICR小鼠为对象,按最大给药剂量经尾静脉注射给药,在4小时、24小时和48小时三个时间点观察小鼠体重、血液学及肝肾功能,电感耦合等离子体质谱(ICP-MS)检测小鼠血液和主要脏器的金纳米杆含量,组织病理学分析金纳米杆对脏器是否损伤,透射电镜确认金纳米杆是否进入组织细胞内。以BALB/C裸鼠为对象,不同浓度金纳米杆、两种给药途径和不同近红外线照射时间作为实验处理因素,观察裸鼠鼻咽癌移植瘤的体积变化、肿瘤增殖率、瘤体病理学及透射电镜表现。结果：本实验构建的PTSB-金纳米杆在近红外区有很强烈的光吸收作用,在生理条件下具有很好的稳定性和生物相容性。静脉给药后,ICR小鼠一般状况良好,体重、血液学及肝肾功能与对照组相比无明显异常,ICP-MS检测发现金纳米杆可经血液循环到达各重要脏器,且肝和脾是其主要蓄积器官,脏器病理学显示无明显损伤,透射电镜确认金纳米杆进入肝、脾和肾细胞内,且大多数出现在溶酶体,少数在细胞质、线粒体。裸鼠鼻咽癌抑瘤实验显示,治疗组瘤体缩小,表面皱缩,随着金纳米杆瘤内给药浓度的增加,近红外线照射时间的增长,治疗组的相对肿瘤增殖率呈下降趋势,肿瘤抑制率呈上升趋势。瘤内给药组与静脉注射给药组相比,同一给药浓度下,瘤内组疗效稍差于静脉组。换算成同一给药剂量下,瘤内组的疗效优于静脉组。照射1.5h和4h疗效优于照射0.5h,但1.5h和4h之间对肿瘤抑制率的影响差别不显著。瘤体病理学显示治疗组肿瘤细胞不同程度变性坏死,肿瘤血管发育差,淋巴细胞浸润程度减轻。透射电镜显示金纳米杆进入肿瘤组织,较多的分布在细胞内的溶酶体,部分散在于细胞质,肿瘤细胞可见凋亡。结论：PTSB-金纳米杆合成过程简便,具有很好的稳定性和生物相容性,是应用于肿瘤红外热疗有前景的纳米材料。PTSB-金纳米杆按最大剂量给药后可随血液循环到达各重要脏器,主要蓄积在肝和脾,对小鼠一般状况和脏器不产生毒性作用,认为可安全地应用于抑瘤实验。PTSB-金纳米杆在红外热疗辅助下对裸鼠鼻咽癌移植瘤有抑制作用,同一给药剂量下,瘤内组的疗效优于静脉组。本实验有望为治疗鼻咽癌提供一种可能的新的治疗方法。
[Abstract]:Objective: low toxicity of nanoparticles is a prerequisite for bioutilization. We attempted to develop a gold nanorods with a novel modified ligand, polymercaptosulfonic betaine polymer (PTSBs), to evaluate the tissue distribution and acute toxicity of PTSB-gold nanorods in mice. To explore the therapeutic effect of PTSB-gold nanorods in nude mice with nasopharyngeal carcinoma (NPC) transplantation assisted by infrared hyperthermia, and to find a new method for the treatment of nasopharyngeal carcinoma (NPC). Materials and methods: PTSB-gold nanorods were constructed by solid combination of sulfhydryl groups and gold nanorods formed by amphoteric ion betaine sulfonic acid (betaine) B). ICR mice were treated by tail vein injection according to the maximum dose. The body weight, hematology, liver and kidney function were observed at four hours, 24 hours and 48 hours, respectively. Inductively coupled plasma mass spectrometry (ICP-MS) was used to detect the content of gold nanorods in blood and main organs of mice. Histopathology was used to analyze whether gold nanorods were damaged or not, and transmission electron microscopy (TEM) was used to confirm whether gold nanorods entered the tissues and cells. BALB/C nude mice with different concentrations of gold nanorods, two ways of administration and different time of near-infrared irradiation were used as experimental factors to observe the volume change and tumor proliferation rate of nasopharyngeal carcinoma xenografts in nude mice. Pathological and transmission electron microscopic findings of the tumor. Results: PTSB-gold nanorods have strong photoabsorption in near infrared region and good stability and biocompatibility under physiological conditions. After intravenous administration, ICR mice were generally in good condition. There was no obvious abnormality in body weight, hematology, liver and kidney function. ICP-MS was used to detect that the hair cash nanorods could reach all important organs through blood circulation, and liver and spleen were the main accumulative organs of ICR mice. The pathology of viscera showed no obvious damage. Transmission electron microscope confirmed that gold nanorods entered into liver spleen and kidney cells and most of them appeared in lysosomes a few in cytoplasm and mitochondria. The tumor inhibition test in nude mice showed that the tumor body in the treatment group was smaller and the surface shrank. With the increase of the concentration of gold nanorods and the increase of the time of near-infrared irradiation, the relative tumor proliferation rate of the treatment group showed a decreasing trend. The tumor inhibition rate was on the rise. Compared with intravenous administration group, the effect of intratumor group was slightly worse than that of intravenous group under the same administration concentration. The curative effect of intratumoral group was better than that of intravenous group under the same dosage. The effect of 1.5 h and 4 h irradiation was better than that of 0.5 h irradiation, but there was no significant difference between 1.5 h and 4 h on tumor inhibition rate. Tumor pathology showed that the tumor cells in the treatment group were denatured and necrotic to varying degrees, the tumor vascular development was poor, and the degree of lymphocytic infiltration was reduced. Transmission electron microscopy (TEM) showed that gold nanorods entered the tumor tissue, and the lysosomes were distributed in the cells, some of which were scattered in the cytoplasm, and apoptosis was observed in the tumor cells. Conclusion the synthesis process of the gold nanorods is simple and has good stability and biocompatibility. It is a promising nanomaterial for tumor infrared hyperthermia. PTSB-gold nanorods can reach various important organs with blood circulation at the maximum dose. It is considered that PTSB-gold nanorods can be safely used in tumor inhibition experiment. PTSB-gold nanorods can inhibit the transplanted tumor of nasopharyngeal carcinoma in nude mice assisted by infrared hyperthermia. The curative effect of intratumoral group was better than that of venous group. This study is expected to provide a new treatment for nasopharyngeal carcinoma.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.63

【参考文献】