本文選題：基因治療 + 雙硫鍵　； 參考：《華中科技大學(xué)》2016年博士論文

【摘要】：第一部分：使用超分子化學(xué)技術(shù)構(gòu)建模塊化自組裝多功能陽(yáng)離子基因傳遞系統(tǒng)目的：構(gòu)建一種基于超分子化學(xué)的主-客體自組裝多功能大分子陽(yáng)離子聚合物基因載體系統(tǒng),評(píng)價(jià)其性能及生物安全性。方法：首先通過對(duì)PCD進(jìn)行修飾,合成了含有雙硫鍵的大分子ATRP引發(fā)劑PCD-SS-BIB;之后通過與DMAEMA的ATRP反應(yīng),得到以聚環(huán)糊精為主鏈、含有雙硫鍵的PDMAEMA為側(cè)鏈的高分子梳狀陽(yáng)離子聚合物PCD-SS-PDMAEMA。通過與金剛烷和葉酸修飾的聚乙二醇(Ad-PEG-FA)自組裝形成超分子聚合物PSD-1/PEG-FA。我們對(duì)聚合物／質(zhì)粒DNA的粒徑、電勢(shì)進(jìn)行了表征以探究聚合物對(duì)質(zhì)粒DNA的壓縮綁定能力；通過在HeLa及COS-7細(xì)胞中的細(xì)胞毒性及溶血實(shí)驗(yàn)檢測(cè)了聚合物的生物相容性；通過熒光素酶報(bào)告基因轉(zhuǎn)染評(píng)價(jià)了聚合物對(duì)質(zhì)粒DNA的轉(zhuǎn)染效果；通過激光共聚焦成像及流式細(xì)胞術(shù)研究了聚合物／質(zhì)粒DNA復(fù)合物的細(xì)胞內(nèi)吞；通過EGFP的siRNA評(píng)價(jià)了聚合物對(duì)siRNA的轉(zhuǎn)染效果；通過葉酸受體陽(yáng)性的KB細(xì)胞驗(yàn)證了葉酸受體的靶向功能。結(jié)果：合成的聚合物可有效地綁定壓縮DNA以及干擾RNA(siRNA)形成納米復(fù)合物。通過針對(duì)該基因載體系統(tǒng)的理化性能表征及相關(guān)生物學(xué)實(shí)驗(yàn)可以證明,該基因載體具有很好的基因綁定壓縮能力、還原敏感性降解能力、低細(xì)胞毒性以及特異性的細(xì)胞吞噬和優(yōu)秀的轉(zhuǎn)染效率。結(jié)論：基于超分子化學(xué)的基因載體合成方法可能為基因載體的“個(gè)性化”生物應(yīng)用提供廣泛的前景。第二部分：CD44靶向的智能響應(yīng)型金-碳納米球治療耐藥性腫瘤目的：構(gòu)建基于空心碳納米球的藥物載體,實(shí)現(xiàn)對(duì)耐藥性腫瘤光熱-光動(dòng)力學(xué)-化學(xué)聯(lián)合治療。方法：首先通過聚多巴胺(PDA)包裹在二氧化硅球的表面形成SiO2@PDA復(fù)合物。之后,通過PDA對(duì)氯金酸鉀的還原,得到SiO2@PDA@Au復(fù)合物。之后,在高溫下碳化PDA并使用氫氟酸去除Si02模板,最終得到了分散有金納米顆粒的金-碳納米球(AuC納米球)。之后通過搭載阿霉素(DOX)及修飾透明質(zhì)酸,得到了聯(lián)合治療納米藥物DOX@AuC@HA。使用紅外成像儀對(duì)金-碳納米球在808nnm近紅外照射下的光-熱轉(zhuǎn)化進(jìn)行了觀察；通過體外釋藥觀察了DOX@AuC@HA對(duì)DOX的釋放;通過血液相容性及細(xì)胞毒性驗(yàn)證了AuC納米球的生物相容性；通過細(xì)胞實(shí)驗(yàn)探究了其體外聯(lián)合治療效果及逆轉(zhuǎn)耐藥性；通過對(duì)MCF-7/ADR荷瘤小鼠的聯(lián)合治療評(píng)價(jià)了其體內(nèi)抗腫瘤效果及體內(nèi)生物相容性。結(jié)果：DOX@AuC@HA具有優(yōu)秀的光熱轉(zhuǎn)化能力、多重敏感的藥物釋放能力以及CD44靶向的細(xì)胞內(nèi)吞。在近紅外光的照射下,DOX@AuC@HA可以通過光熱和光動(dòng)力學(xué)的效果極大的促進(jìn)其所包載的DOX的療效。在體內(nèi)實(shí)驗(yàn)中,我們也證實(shí)了DOX@AuC@HA可以通過系統(tǒng)給藥有效的分布到腫瘤部位,且顯示出了非常理想的抗腫瘤能力。結(jié)論：多功能、多治療策略聯(lián)用的DOX@AuC@HA納米藥物可能在治療耐藥性腫瘤上發(fā)揮巨大的作用。
[Abstract]:Part I: construction of modular multifunctional cationic gene transfer system based on supramolecular chemistry objective: to construct a host-guest self-assembly multi-functional macromolecular cationic polymer gene vector system based on supramolecular chemistry Its performance and biological safety were evaluated. Methods: the macromolecular ATRP initiator PCD-SS-BIBB containing disulfide bond was synthesized by modifying PCD, and then the comb-like cationic polymer PCD-SS-PDMAEMAA was obtained by reacting with the ATRP of DMAEMA and PDMAEMA with PDMAEMA as side chain. A supramolecular polymer PSD-1 / PEG-FAA was synthesized by self-assembly of polyethylene glycol Ad-PEG-FAA modified with adamantane and folic acid. The particle size and potential of polymer / plasmid DNA were characterized to investigate the binding ability of polymer to plasmid DNA, and the biocompatibility of polymer was tested by cytotoxicity and hemolysis test in HeLa and COS-7 cells. The transfection effect of polymer on plasmid DNA was evaluated by luciferase reporter gene transfection, and the endocytosis of polymer / plasmid DNA complex was studied by confocal laser imaging and flow cytometry. The transfection effect of the polymer on siRNA was evaluated by EGFP siRNA, and the targeted function of folate receptor was verified by folate receptor positive KB cells. Results: the synthesized polymers can effectively bind to compress DNA and interfere with RNA-siRNAs to form nanocomposites. According to the physical and chemical properties of the gene vector system and related biological experiments, it can be proved that the gene vector has a good ability of gene binding compression and reduction sensitivity degradation. Low cytotoxicity, specific phagocytosis and excellent transfection efficiency. Conclusion: the method of gene vector synthesis based on supramolecular chemistry may provide a broad prospect for individualized biological application of gene vector. Part II: intelligent responsive gold-carbon nanospheres targeting CD44 for the treatment of drug-resistant tumors objective: to construct a drug carrier based on hollow carbon nanospheres and to achieve photothermal-photodynamic chemotherapeutic combined therapy for drug-resistant tumors. Methods: at first, Sio _ 2 @ PDA complex was formed on the surface of silica sphere by polydopamine PDA-encapsulated. After that, through the reduction of potassium chloraurate by PDA, the Sio _ 2 @ PDA-R au complex was obtained. After carbonation of PDA at high temperature and removal of Si02 template with hydrofluoric acid, au C nanospheres dispersed with gold nanoparticles were obtained. After that, the combination therapy of doxobenzene (DOX) and modified hyaluronic acid (HA) was obtained. The photothermal transformation of gold-carbon nanospheres irradiated by 808nnm near infrared irradiation was observed by infrared imager, the release of DOX from DOXR was observed by drug release in vitro, and the biocompatibility of AUC nanospheres was verified by blood compatibility and cytotoxicity. The in vitro combined treatment and reversal of drug resistance were investigated by cell experiments, and the anti-tumor effect and biocompatibility of MCF-7 / ADR bearing mice were evaluated. Results the results showed that: 1. The ability to convert photoheat, the ability of drug release and the endocytosis of CD44 were excellent. DOX @ AuC@ HA can greatly promote the efficacy of DOX by photothermal and photodynamic effects under near-infrared light irradiation. In vivo experiments, we also confirmed that DOX @ AuC@ HA can be effectively distributed to the tumor site through systemic administration, and has shown a very good anti-tumor ability. Conclusion: the multifunctional, multitherapy strategy combined with DOX @ AuC@ HA nanopharmaceuticals may play an important role in the treatment of drug-resistant tumors.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R730.5

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 趙海全;沈家成;劉蕊;;石油庫(kù)埋地管道外防腐層涂料的選材分析[J];石油庫(kù)與加油站;2012年05期

2 冉龍飛;王曉霖;;含體積型缺陷管道的剩余強(qiáng)度評(píng)價(jià)[J];石油化工高等學(xué)校學(xué)報(bào);2015年01期

3 劉斌;;天然氣管道腐蝕監(jiān)測(cè)[J];化工管理;2014年24期

4 張鵬;趙國(guó)仙;畢宗岳;李鴻斌;王軍;趙勇;;油氣管道內(nèi)防腐涂層性能研究[J];焊管;2014年01期

5 李欣;李進(jìn);;長(zhǎng)輸天然氣氣質(zhì)對(duì)管道內(nèi)腐蝕的影響研究[J];石油化工高等學(xué)校學(xué)報(bào);2015年02期

6 馬曉紅;常大勝;;長(zhǎng)嶺至松原含CO_2天然氣管道腐蝕監(jiān)測(cè)與防護(hù)[J];中小企業(yè)管理與科技(上旬刊);2015年05期

7 張偉剛;趙會(huì)軍;周立輝;程雅雯;彭浩平;;濕氣管線的頂部腐蝕研究進(jìn)展[J];腐蝕科學(xué)與防護(hù)技術(shù);2015年05期

8 雷曉曾;張陽(yáng);;天然氣長(zhǎng)輸管道運(yùn)行管理中易發(fā)問題及綜合治理淺析[J];內(nèi)蒙古石油化工;2015年15期

9 張桉;黎庶;;礦化度對(duì)油氣管道腐蝕的研究[J];石化技術(shù);2016年07期

10 張偉剛;趙會(huì)軍;張璇;程雅雯;;X80管線鋼在含硫原油中的頂部腐蝕行為[J];腐蝕與防護(hù);2016年08期

，

本文編號(hào)：2048793