论文总字数：19834字

摘 要

背景：含铁纳米粒在肿瘤高表达过氧化氢时可以诱导产生自由基，促进肿瘤死亡，达到治疗肿瘤的治疗效果。本论文基于单宁酸和聚醚F-127之间的多价氢键结合聚(环氧丙烷)链的疏水相互作用，利用铁离子和多羟基单宁酸的螯合作用，构建铁死亡纳米粒并对其制备工艺进行优化。

方法和结果：对PPNP-Fe进行的粒径检测，纳米体系对铁的包封率检测，MB显色实验，TMB显色实验，PPNP-Fe体系中铁的释放率测定和在酸性条件下对铁氧化循环的验证相关实验。本实验采用邻菲罗琳法与紫外分光光度法相结合的方法实现对PPNP-Fe中微量铁的测定，结果表明纳米体系对铁的包封率为20.27%。优化后的纳米体系97%以上的粒径在65-220nm之间并且30h内粒径稳定；在对PPNP-Fe释放率进行相关测定的实验中，实验结果表明在pH=7.4时PPNP-Fe几乎不释放铁离子，随着pH值的下降，纳米体系铁的释放率会有所提升，在pH=4.0醋酸盐缓冲溶液中，温度为27℃时，20ml的24h的释放率可达到50.39%。用MB显色实验对铁死亡反应中的自由基进行检测，显现出较为明显的结果。用TMB实验对铁离子的催化能力进行检测，也显示出较好的催化能力。

结论：PPNP-Fe纳米体系存在金属酚醛网，可用来包载治疗药物，因此其在联合铁死亡和其他治疗对癌症进行治疗上有着比较广阔的应用前景。

关键词：单宁酸 铁死亡 TMB显色实验 MB显色实验 邻菲罗啉

Construction of  Ferroptosis nanometer and its in vitro evaluation

Abstract

Background: iron-containing nanoparticles can induce the generation of free radicals, promote the death of tumors, and achieve the therapeutic effect of treating tumors when hydrogen peroxide is highly expressed in tumors.In this paper, based on the hydrophobic interaction between tannic acid and polyether f-127 (propylene oxide) chain, iron death nanoparticles were constructed and their preparation process was optimized by using the chelation of iron ions and polyhydroxytannic acid.

Methods and results: particle size detection of ppnp-fe, encapsulation rate detection of iron in nanometer system, MB color rendering experiment, TMB color rendering experiment, determination of iron release rate in ppnp-fe system and verification of iron oxidation cycle under acidic conditions.In this experiment, the determination of trace iron in ppnp-fe was carried out by the method of o-philocline and ultraviolet spectrophotometry.The particle size of the optimized nanometer system was more than 97% between 65-220nm and was stable within 30h.In the experiment to determine the release rate of ppnp-fe, the experimental results showed that at pH=7.4, ppnp-fe almost did not release iron ions, and with the decrease of pH value, the release rate of iron in the nanometer system would be improved. In the buffer solution of pH=4.0 acetate, when the temperature was 27℃, the release rate of 20ml 24h could reach 50.39%.MB chromogenic experiment was used to detect the free radicals in iron death reaction.The catalytic capacity of iron ions was tested by TMB experiment, which also showed good catalytic capacity.

Conclusion: the ppnp-fe nano system has a metal phenolic mesh, which can be used to package therapeutic drugs, so it has a broad application prospect in the treatment of cancer combined with iron death and other treatments.

Keywords： Tannins acid;Ferroptosis;TMB;MB; Phenanthroline

目 录

摘要................................................................................I

Abstract II

第一章 绪 论 3

1.1 本课题的背景和意义 3

1.2 芬顿反应和铁死亡 3

1.3 装载三价铁的纳米载体的选择及原理 4

第二章 PPNP-Fe纳米载体制备 5

2.1 实验所需仪器、材料 5

2.2 PPNP-Fe的制备工艺 6

2.3 PPNP-Fe的工艺优化过程以及粒径检测和粒径稳定性考察 6

第三章 PPNP-Fe的体外指标的检测 8

3.1 PPNP-Fe中Fe包载量的测定 8

3.2 MB显色实验 11

3.2.1 PPNP-Fe进行MB显色实验的实验原理 11

3.2.2 MB显色实验的问题和解决方法 11

3.2.3 PPNP-Fe的MB显色随时间变化的实验 12

3.2.4 PPNP-Fe的浓度梯度MB显色实验 12

3.3 TMB显色实验 13

3.3.1 TMB的结构和性质 13

3.3.2 PPNP-Fe的TMB预实验出现的问题 14

3.3.3 PPNP-Fe的TMB显色实验随时间变化曲线 16

3.3.4 PPNP-Fe的TMB显色实验的浓度梯度变化曲线 16

3.3.5实验结果不为蓝色的原因 17

3.4 PPNP-Fe纳米体系铁离子的释放 18

3.4.1 实验步骤 18

3.4.2 实验结果 18

3.4.3结论 19

3.5 酸性条件下对铁氧化循环的验证试验 19

3.5.1 实验步骤 19

3.5.2 实验结果 20

结 论 21

参考文献 22

致谢 24

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