【作者】 刘欢；

【导师】 肖恩华；

【作者基本信息】 中南大学 ， 临床医学（专业学位）， 2023， 硕士

【摘要】 目的：基于白蛋白结合策略,创新性合成一种新型光敏剂EB-Ppa,用于增强肿瘤靶向成像和光动力治疗,以实现肿瘤的诊疗一体化。方法：构建分子探针EB-Ppa并采用紫外分光光度计、质谱仪进行吸收光谱和发射光谱以及质谱检测;然后采用RNO-ID实验检测EB-Ppa活性氧的生成;采用CCK-8试剂盒评价细胞毒性及体外光动力治疗实验;利用Kodak In Vivo FX小动物光学成像仪检测探针的体内成像能力和生物分布;采用4T1瘤鼠进行体内光动力治疗实验,H&E染色、TUNEL凋亡染色及PCNA增殖染色用于评估光动力治疗对肿瘤的杀伤作用;最后对光动力治疗后的4T1瘤鼠主要器官进行H&E染色和血清生化分析,用于评价探针的生物安全性。结果：相比于游离Ppa,EB-Ppa与白蛋白结合后,RNO吸光度最高下降了15%,而游离Ppa的吸光度仅下降了6%;其次正常光照条件下,EB-Ppa在200μg m L^-1的最高浓度下,其细胞活力也高于90%。而当用光动力治疗10分钟后,其细胞活力显著下降至10%以下。瘤鼠尾静脉注射200μg m L^-1浓度的EB-Ppa探针4小时后,EB-Ppa开始在肿瘤区域的逐渐聚集,并在24小时后肿瘤区域的积聚达到顶峰。之后进行激光诱导的光动力治疗,实验组瘤鼠的肿瘤体积从基线的155.7mm³下降至11.6mm³,而其余三组体积增长至少达初始值的4倍以上。最后主要器官（心、肝、脾、肺和肾）的H&E染色和血清生化指标显示均无明显不良反应和急性损伤。结论：本文成功构建了一种新型的白蛋白结合光敏剂EB-Ppa,其具有较高的活性氧生成率、良好的肿瘤靶向能力及生物相容性,能够有效用于4T1肿瘤的靶向成像和光动力治疗。图11幅,表格2个,参考文献145篇更多还原

【Abstract】 Objective:Based on the albumin binding strategy,a new type of albumin binding photosensitizer agent EB-Ppa was innovatively synthesized for enhanced tumor targeted imaging and PDT therapy to realize the integration of tumor diagnosis and treatment.Methods:The molecular probe EB-Ppa was constructed and detected by UV/Vis spectrometer and mass spectrometer;Then,RNO-ID experiment was used to detect the generation of EB-Ppa reactive oxygen species;The CCK-8 kit was used to evaluate cytotoxicity and in vitro PDT experiments;The in vivo imaging ability and biodistribution of the probe were detected by the Kodak In Vivo FX small animal optical imager;In vivo PDT experiments using 4T1 tumor-bearing nude mice,H&E staining,TUNEL staining and PCNA staining were used to evaluate the killing effect of PDT;Finally,H&E staining and serum biochemical analysis of the major organs of 4T1 tumor-bearing nude mice after PDT treatment was used to evaluate the biological safety of the probes.Result:Compared with free Ppa,the absorbance of RNO decreased by 15%,while the absorbance of free Ppa decreased by only 6%.Secondly,the cell viability of EB-Ppa was higher than 90%at the highest concentration of 200μg m L^-1.However,when treated with PDT for 10minutes,its cell viability significantly decreased to less than 10%.Then four hours after injection of 200μg m L^-1probe,the gradual accumulation of EB-Ppa was initiated in the tumor region and peaked in the accumulation in the tumor region 24 hours later.After laser-induced photodynamic treatment,the tumor volume of the experimental group of tumors decreased from 155.7mm³at baseline to 11.6mm³,while the remaining three groups increased by at least four times the initial value.H&E staining and serum biochemical parameters of the major organs（heart,liver,spleen,lung and kidney）showed no significant adverse effects and acute injury.Conclusion:A novel albumin-binding photosensitizer EB-Ppa was successfully constructed in this thesis,which has high reactive oxygen species generation rate,tumor targeting ability and good biocompatibility,and can be effectively used for targeted imaging and PDT treatment of4T1 tumors.更多还原