纳米金刚石载药及其对HL-60细胞凋亡作用的研究

发布时间：2018-06-16 11:42

本文选题：纳米金刚石 + 表面修饰　；参考：《吉林大学》2014年博士论文

【摘要】：纳米金刚石具有非常优异的物理、化学特性，特别是其生物相容性及功能化可修饰性，使纳米金刚石在生物、医学等诸多领域具有非常重要的研究价值和应用前景。纳米金刚石的引入，可以提高载药效果，并应用于多种肿瘤细胞的化疗。进一步拓展纳米金刚石对不同药物的载药及提高药物的治疗效果，成为近年来纳米材料在生物医学研究领域的重要前沿课题。本论文以爆炸法纳米金刚石(DND)为基础，通过表面功能化，连接全反式维甲酸(ATRA)，研究其在人早幼粒白血病细胞(HL-60细胞)凋亡过程中的促进作用。主要研究内容和结果如下： 1.采用超声、高温等处理过程，通过相转变和氧化反应，，以及醇分子与纳米金刚石液相化学反应，对平均粒度约5nm的爆炸法纳米金刚石(DND)进行颗粒分散。实验表明，相对于其他方法，液相反应使纳米金刚石表面修饰了酯类化学基团，基本呈单分散状态。 2.将用于治疗急性早幼粒白血病(APL)的ATRA药物与单分散的表面修饰纳米金刚石颗粒相结合，纳米金刚石表面修饰的酯类基团的疏水性有利于ATRA的碳环端与纳米金刚石修饰表面之间的吸附结合，制备了DND-ATRA纳米复合物，通过傅立叶变换红外光谱、紫外-可见荧光光谱和透射电子显微镜等测量，证明了ATRA和纳米金刚石有效地组装在一起，可用于提高ATRA有效浓度和利用率。 3.利用四甲基偶氮唑蓝比色法(MTT)、流式细胞仪检测(FACS)及原位末端标记(TUNEL)荧光镜检等检测方法，我们系统研究了DND-ATRA纳米复合药物对人早幼粒白血病细胞HL-60细胞的体外促凋亡作用。DND-ATRA复合物作用2天后，MTT结果显示HL-60细胞的抑制率与单独使用ATRA相比提高了38%，并且在8天药物作用后，DND-ATRA纳米复合物实验组达到93%的HL-60有效的凋亡率。与常规的单独ATRA或者其他ATRA-化疗药物作用方式相比，DND-ATRA纳米复合物体系不仅保持了ATRA本身诱导分化癌细胞的药物活性，而且更有效地促进了HL-60癌细胞凋亡，较ATRA的细胞凋亡率提高了35%，细胞水平上证明了DND-ATRA纳米复合物的有效性，同时说明，纳米金刚石是优秀的生物相容性载体，可抑制细胞膜转运蛋白对药物分子的外排效应，并对细胞没有破坏作用。我们认为单纯ATRA的代谢形式为胞内直接扩散，易于降解和清除，而纳米复合物具有细胞局部区域扩散累积效应，可比单独ATRA能更持久地诱导HL-60细胞分化凋亡。本论文研究表明，DND-ATRA的纳米复合物可以成为高效治疗人早幼粒白血病的潜力的药剂，证明了以纳米金刚石为基础，所构建的新型药物输送体系具有很好的可行性和实用性，为今后的动物实验以及纳米金刚石药物输送载体对各种功能基团和药物分子的负载，提供了一定的实验数据和工作基础，具有潜在的临床应用意义。
[Abstract]:Nano diamond has very excellent physical and chemical properties, especially its biocompatibility and functionalization modifiable, which makes nano diamond have very important research value and application prospect in many fields, such as biology, medicine and so on. The introduction of nano diamond can improve the effect of drug carrying and apply to chemotherapy of various tumor cells. One step to expand the effect of nano diamond on drug loading and drug treatment has become an important frontier topic in the biomedical research field in recent years. This paper, based on the nano diamond (DND), is based on the surface functionalization and connection of all trans retinoic acid (ATRA) to study its fine promyelocytic leukemia. The promoting effect of cell (HL-60 cell) in the process of apoptosis. The main contents and results are as follows:
1. in the process of ultrasonic and high temperature treatment, by phase transition and oxidation reaction, and the chemical reaction between alcohol molecules and nano diamond liquid, the particle dispersion of the nano diamond (DND) with an average particle size of 5nm is dispersed. The experiment shows that the liquid phase reaction makes the nano diamond surface modified with the chemical groups of the esters relative to the other methods. It is monodisperse.
2. the ATRA drugs used in the treatment of acute promyelocytic leukemia (APL) were combined with monodisperse surface modified nano diamond particles. The hydrophobicity of the nano diamond surface modified ester groups was beneficial to the adsorption of the carbon ring ends of ATRA and the nano diamond modified surface. The DND-ATRA nanocomposites were prepared by Fu Liye change. Infrared spectroscopy, ultraviolet visible fluorescence and transmission electron microscopy have proved that ATRA and nano diamond are effectively assembled together, which can be used to improve the effective concentration and utilization of ATRA.
3. using four methyl azolazolium Colorimetry (MTT), flow cytometer detection (FACS) and in situ terminal labeling (TUNEL) fluoroscopy, we systematically studied the effect of DND-ATRA nanocomposite on the apoptosis of HL-60 cells of human promyelocytic leukemia cells in vitro for 2 days, and MTT results showed HL-60 cells. The inhibition rate increased by 38% compared with the single use of ATRA, and after 8 days of drug action, the DND-ATRA nanocomposite experimental group reached 93% of the effective apoptosis rate of HL-60. Compared with the conventional single ATRA or other ATRA- chemotherapeutic agents, the DND-ATRA nanocomposite system did not only maintain the drug of ATRA itself to induce the differentiation of cancer cells. Activity, and more effective to promote the apoptosis of HL-60 cancer cells, increased the apoptosis rate of ATRA by 35%. The cell level proved the effectiveness of DND-ATRA nanocomposites. At the same time, the nano diamond is an excellent biocompatibility carrier, which can inhibit the efflux of cell membrane transporter to drug molecules and not to cells. We think that the metabolic form of simple ATRA is intracellular direct diffusion, which is easy to degrade and clear, and the nanocomposite has the effect of local area diffusion accumulation, which can induce HL-60 cells to differentiate and apoptosis more consistently than single ATRA.
This study shows that DND-ATRA nanocomposite can be a potent agent for the treatment of human promyelocytic leukemia. It is proved that the new drug delivery system based on nano diamond has good feasibility and practicability. It can be used for the future animal experiment and the carrier of nano diamond drug delivery. The loading of energy groups and drug molecules provides certain experimental data and work basis, which has potential clinical application significance.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：TB383.1;R943;R96

【参考文献】