叶酸修饰的和厚朴酚主动靶向药物对鼻咽癌的抑制作用与机制研究

发布时间：2018-06-25 07:49

本文选题：和厚朴酚 + 鼻咽癌　；参考：《西南医科大学》2017年硕士论文

【摘要】：目的:制备并验证叶酸修饰的靶向人鼻咽癌HNE-1细胞的和厚朴酚的纳米药物系统,并探讨其对鼻咽癌抑制效果和机制。方法:本文使用两亲性三嵌段聚合物:聚己内酯-聚乙二醇-聚己内酯(PCL-PEG-PCL,PCEC)为原材料,采用聚醚酰亚胺(PEI)引入胺基后,与叶酸(Folate,FA)共价结合,制备了目标载体聚合物PCEC-PEI-FA。然后,以PCEC-PEI-FA聚合物为载体材料,通过薄膜分散法自主装负载和厚朴酚形成具有主动靶向功能的载药纳米粒(ATNH),并使用透射电镜、激光粒度仪、红外光谱仪及核磁氢谱考察纳米聚合物的形貌、粒径及结构,溶血实验考察溶血安全性、透析法考察体外释药特性。使用噻唑蓝(methylthiazoletetrazolium, -MTT)试验及荧光显微镜分别研究了该纳米粒的细胞毒性和主动靶向作用。建立人鼻咽癌裸鼠体内皮下移植瘤模型,并接受主动靶向纳米粒的治疗。肿瘤再生长延迟时间曲线和中位生存期用于评价载药纳米粒的抗肿瘤疗效。18F-脱氧葡萄糖(fluorine-18-deoxyglucose,FDG) PET/CT显像用于评价肿瘤对载药纳米粒的早期反应。此外,载药纳米粒的抗肿瘤机制研究是通过流式细胞仪检测细胞周期分布与细胞凋亡和免疫组化检测血小板-内皮细胞黏附分子(CD 31)反应抗血管生成作用及增殖细胞核抗原(Ki 67)反应抗增殖作用。结果:该研究制备的主动靶向功能的和厚朴酚纳米粒(ATNH)呈球形,平均流体动力学直径为188.34 ±0.54 nm,无明显的突发释放,并呈现出缓慢释药的特点,未发生溶血现象。MTT实验结果显示:主动靶向载药纳米粒(ATNH)对肿瘤细胞的细胞毒性明显高于普通和厚朴酚,其作用机制可能与叶酸受体(folate receptor, FR)介导的细胞内吞作用有关。主动靶向载药纳米(ATNH)组的细胞凋亡指数为86.07 ±4.95%,明显高于各对照组(P 0.01).体内研究结果表明,主动靶向载药纳米粒(ATHN)组的肿瘤延迟生长时间及中位生存时间分别为24天及57.5天,明显高于普通和厚朴酚对照组(肿瘤延迟生长时间及中位生存时间分别为8天及34天)及非主动靶向纳米组(肿瘤延迟生长时间及中位生存时间分别为18天及42.5天)(P 0.01 )。小动物PET/CT显像结果示主动靶向载药纳米粒(ATHN)组的T/M值为1.03±0.42(P0.01)。免疫组化结果显示:主动靶向载药纳米粒(ATHN)组的Ki 67阳性指数为37.24 ± 3.89% (P 0.01),CD31阳性指数为2.16±0.76(P0.01)。与对照组相比,主动靶向载药纳米粒(ATHN)组在G1期的细胞比例升高,为51.24 ± 3.95%,明显高于对照组(p 0.01)。结论:叶酸修饰的和厚朴酚纳米粒具有对鼻咽癌细胞(HNE-1)的主动特异性靶向功能,对鼻咽癌移植瘤具有非常明显的抗肿瘤作用。其抗癌机制可能与诱导人鼻咽癌细胞凋亡,G1期细胞周期阻滞,抑制肿瘤细胞增殖、肿瘤血管生成,并抑制肿瘤代谢有关。表明该载药纳米粒对于治疗鼻咽癌等细胞表面叶酸受体表达丰富的肿瘤具有潜在的运用前景。
[Abstract]:Objective: to prepare and verify the nanoscale drug system of HNE-1 cells and magnolol in human nasopharyngeal carcinoma cells modified by folic acid, and to explore its inhibitory effect and mechanism on nasopharyngeal carcinoma. Methods: This paper uses polyhexyl polyhexyl polyhexyl polyhexyl (PCL-PEG-PCL, PCEC) as the raw material and the introduction of polyether imide (PEI) in the study of the inhibition of nasopharyngeal carcinoma (HNE-1). After the amine group, the target carrier polymer PCEC-PEI-FA. was prepared by covalent binding with folic acid (Folate, FA). Then, the PCEC-PEI-FA polymer was used as the carrier material. The load and the active targeting drug loaded nanoparticles (ATNH) were formed independently by the thin film dispersion method and the magnolol was formed by the thin film dispersion method. The transmission electron microscope, laser particle size meter, infrared spectrometer and nuclear magnetic hydrogen were used. The morphology, particle size and structure of nanoscale, hemolysis safety and drug release in vitro were investigated by hemolytic test. The cytotoxicity and active targeting effect of the nanoparticles were studied by methylthiazoletetrazolium (-MTT) test and fluorescence microscopy. The subcutaneous transplantation tumor in nude mice of human nasopharyngeal carcinoma was established. The model, and the treatment of active targeting nanoparticles. The tumor regrowth delay time curve and the median survival time are used to evaluate the antitumor effect of drug loaded nanoparticles,.18F- deoxyglucose (fluorine-18-deoxyglucose, FDG) PET/CT imaging for the evaluation of the early response of tumor to drug loaded nanoparticles. In addition, the anti-tumor mechanism of drug loaded nanoparticles The study was to detect the anti angiogenesis effect of cell cycle distribution, cell apoptosis and immunohistochemical detection of platelet endothelial cell adhesion molecule (CD 31) in response to angiogenesis and proliferating cell nuclear antigen (Ki 67). Results: the active targeting function and the magnolol nanoparticles (ATNH) were spherical and flat. The average fluid dynamic diameter was 188.34 + 0.54 nm, and there was no obvious burst release and showed the characteristics of slow release. The result of.MTT experiment without hemolysis showed that the cytotoxicity of active target drug loaded nanoparticles (ATNH) to tumor cells was obviously higher than that of common and magnolol. The mechanism of its action may be related to the folic acid receptor (folate receptor, FR). The apoptosis index of the active targeted drug loaded nanoparticles (ATNH) was 86.07 + 4.95%, which was significantly higher than that of the control group (P 0.01). In vivo studies showed that the delayed growth time and the median survival time of the active targeted drug loaded nanoparticles (ATHN) group were 24 days and 57.5 days respectively, which were significantly higher than those of ordinary and Magnolia officinalis. The phenol control group (the delayed growth time and the median survival time of the tumor was 8 and 34 days respectively) and the non active target group (18 days and 42.5 days, respectively, for the delayed growth time and the median survival time of the tumor) (P 0.01). The PET/CT imaging results of the small animals showed that the T/M value of the active targeted drug loaded nanoparticles (ATHN) group was 1.03 + 0.42 (P0.01). The results showed that the positive index of Ki 67 in active targeted drug loaded nanoparticles (ATHN) was 37.24 + 3.89% (P 0.01) and CD31 positive index was 2.16 + 0.76 (P0.01). Compared with the control group, the proportion of active targeted drug loaded nanoparticles (ATHN) in G1 phase increased, which was 51.24 + 3.95%, obviously higher than that of the control group (P 0.01). Conclusion: folic acid modified and Magnolia officinalis. Phenolic nanoparticles have an active specific targeting function to nasopharyngeal carcinoma cells (HNE-1), which have an obvious antitumor effect on nasopharyngeal carcinoma transplantation tumor. Its anticancer mechanism may be related to inducing apoptosis of human nasopharyngeal carcinoma cells, cell cycle arrest in G1 phase, inhibition of tumor cell proliferation, tumor angiogenesis, and inhibition of tumor metabolism. Nanoparticles have potential applications in the treatment of nasopharyngeal carcinoma and other tumor cells with abundant folate receptors.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.63

【相似文献】