环状有机铋（锑）配合物生物活性的研究

发布时间：2019-06-18 17:28

【摘要】：铋、锑具有悠久的药用历史。近期研究表明铋、锑配合物具有良好的抑菌和抗肿瘤等生物活性。但由于常规的有机铋(锑)配合物对水和空气较为敏感，稳定性差，其应用受到限制。因此，设计开发具有较高稳定性的有机铋(锑)配合物，并探索其在生命科学中的应用，具有重要的学术意义和应用价值。本文以配合物与DNA分子的作用方式为依据，选取具有共轭芳香环的有机铋(锑)配合进行研究。利用紫外-可见吸收光谱谱学手段，证实环状有机铋(锑)配合物与小牛胸腺DNA (CT-DNA)以插入方式相互作用。以人肺腺癌细胞A549、肝癌细胞HepG2、食管癌Ec109等细胞为模型，采用CCK-8法从配合物中筛选出能有效抑制肿瘤细胞增殖的C3([t-BuN(CH2C6H4)2]BiCl)、C4([O(CH2C6H4)2]BiCl)、C5([O(CH2C6H4)2]BiNO3)、 C9([t-BuN(CH2C6H4)2]SbCl)和C12([{t-BuN(CH2C6H4)2}Sb]2O)。构效关系研究结果表明配合物的抗增殖活性可能与铋、锑和配位原子之间的配位作用有关。为了进一步研究配合物的抗增殖机制，本文采用了流式细胞术检测配合物对细胞周期和细胞凋亡的影响，使用激光共聚焦显微镜观察配合物作用下A549细胞的形态学变化及凋亡现象，利用realtime PCR及Western Blot分析凋亡相关基因和蛋白的表达。一系列的实验结果表明，，环状有机铋配合物C3、C4、C5和环状有机锑配合物C9、C12对细胞周期的阻滞作用可能与其抗肿瘤细胞增殖效应有关，另一方面，配合物可能通过Bcl-2表达的下调及Bax的过表达诱导的细胞凋亡。本文采用试管法和琼脂扩散法考察环状有机铋(锑)配合物的抑菌作用，研究表明与铋、锑配位基团具有较小位阻和较强供电能力的环状有机铋配合物(C3([t-BuN(CH2C6H4)2]BiCl)、C4([O(CH2C6H4)2]BiCl)、C5([O(CH2C6H4)2]BiNO3))和环状有机锑配合物(C9([t-BuN(CH2C6H4)2]SbCl)和C12([{t-BuN(CH2C6H4)2}Sb]2O)同样具有较好抑菌活性，尤其是对革兰氏阳性菌的抑菌作用呈剂量依赖性。本研究成果为深入探究环状有机铋(锑)的抗肿瘤及抑菌作用机制，为生物活性分子的合理设计、活性化合物的筛选以及药物的研制提供理论及实验依据。
[Abstract]:Bismuth and antimony have a long medicinal history. Recent studies have shown that bismuth and antimony complexes have good bacteriostatic and antitumor activities. However, due to the sensitivity and poor stability of conventional organic bismuth (antimony) complexes to water and air, their application is limited. Therefore, it is of great academic significance and application value to design and develop organic bismuth (antimony) complexes with high stability and to explore their application in life science. Based on the interaction of complexes with DNA molecules, the coordination of organic bismuth (antimony) with conjugated aromatic rings was studied in this paper. The interaction between cyclic organic bismuth (antimony) complex and calf thymic DNA (CT-DNA) was confirmed by UV-vis absorption spectroscopy. C3 ([t-BuN (CH2C6H4) 2] BiCl), C 4 ([O (CH2C6H4) 2] BiCl), C 5 (O (CH2C6H4) 2] BiNO3), C9 ([t-BuN (CH2C6H4) 2] SbCl) and C12 ([{t-BuN (CH2C6H4) 2} Sb] 2O) were screened from human lung adeno cell line A549 and hepatocellular carcinoma cell line HepG2, esophageal carcinoma Ec109 by CCK- 8 method. The results of structure-activity relationship show that the anti-proliferation activity of the complex may be related to the coordination between bismuth, antimony and coordination atoms. In order to further study the anti-proliferation mechanism of the complex, the effects of the complex on cell cycle and apoptosis were detected by flow cytometry. The morphological changes and apoptosis of A549 cells under the action of the complex were observed by laser confocal microscope, and the expression of apoptosis-related genes and proteins was analyzed by realtime PCR and Western Blot. A series of experimental results show that the blocking effects of cyclic organic bismuth complexes C _ 3, C _ 4, C _ 5 and cyclic organic antimony complexes C _ 9 and C _ 12 on cell cycle may be related to their anti-tumor cell proliferation effects. on the other hand, the complexes may induce apoptosis through down-regulation of Bcl-2 expression and overexpression of Bax. The bacteriostatic effect of cyclic organic bismuth (antimony) complex with bismuth and antimony was investigated by tube test method and Agar diffusion method. The results showed that cyclic organic bismuth complex (C3 ([t-BuN (CH2C6H4) 2] BiCl), C 4 ([O (CH2C6H4) 2] BiCl),) with bismuth and antimony coordination group had small steric hindrance and strong power supply ability. C5 ([O (CH2C6H4) 2] BiNO3) and cyclic organic antimony complexes (C9 ([t-BuN (CH2C6H4) 2] SbCl) and C12 ([{t-BuN (CH2C6H4) 2} Sb] 2O) also have good bacteriostatic activity, especially against Gram-positive bacteria in a dose-dependent manner. The results of this study provide theoretical and experimental basis for the anti-tumor and bacteriostatic mechanism of cyclic organic bismuth (antimony), and for the rational design of bioactive molecules, the screening of active compounds and the development of drugs.
【学位授予单位】：湖南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96

【参考文献】