本文選題：氧化石墨烯（GO） + 聚乙烯亞胺（PEI）��； 參考：《吉林大學(xué)》2015年碩士論文

【摘要】：基因治療對(duì)于治療疾病相關(guān)，尤其是癌癥是一個(gè)熱點(diǎn)�；蛑委熥铌P(guān)鍵的問(wèn)題就是發(fā)展一個(gè)合適、高效的基因載體，基因載體可分為病毒型載體和非病毒型載體。病毒型載體由于毒性高和自身的局限性而使用的少，非病毒載體因?yàn)榘踩愿摺⒂行詮?qiáng)得到了廣泛的應(yīng)用，例如：聚合物、脂質(zhì)體、納米粒子等等。近幾年，非病毒基因載體雖然已經(jīng)獲得了很大的進(jìn)步，但有些問(wèn)題包括非特異性靶向位點(diǎn)、毒性和生物穩(wěn)定等問(wèn)題依然沒(méi)有得到很好的解決。因此發(fā)展一個(gè)有效的納米載體仍然是基因治療中最大的挑戰(zhàn)。 石墨烯，擁有一個(gè)碳原子厚的二維層狀結(jié)構(gòu)，因?yàn)檫@個(gè)特異性在許多領(lǐng)域中都得到了廣泛的應(yīng)用，例如電化學(xué)裝置，生物應(yīng)用等等。氧化石墨烯(GO)是石墨烯的水溶性衍生物，由于其容易制備，在水溶液和生理環(huán)境中有較好的分散性，好的膠體穩(wěn)定性已經(jīng)吸引了很大的興趣，并且在生物傳感器，基因和藥物傳遞，以及在體內(nèi)外的生物成像等生物領(lǐng)域中也顯示了潛在的應(yīng)用。 在本文中，我們以氧化石墨烯為基礎(chǔ)，采用對(duì)其進(jìn)行化學(xué)偶聯(lián)修飾，設(shè)計(jì)一種新型的多功能氧化石墨烯的基因載體GO-PEI-PEG-FA/si-Stat3。我們選擇常用的聚合物支化的聚乙烯亞胺（PEI）與羧基化的氧化石墨烯通過(guò)偶氮化學(xué)反應(yīng)進(jìn)行連接，目的是為提高轉(zhuǎn)染效率。引入了聚乙二醇（PEG），為提高溶液的生物穩(wěn)定性，最后引入一種小分子葉酸（FA），對(duì)其進(jìn)行靶向性修飾，實(shí)現(xiàn)靶向治療的目的。再與si-Stat3質(zhì)粒通過(guò)靜電相互作用與GO-PEI-PEG-FA連接。最終形成GO-PEI-PEG-FA/si-Stat3基因載體。 我們研究了通過(guò)調(diào)節(jié)不同比例的PEI對(duì)GO-PEI-PEG-FA的結(jié)構(gòu)的影響，并在這個(gè)過(guò)程進(jìn)一步研究結(jié)構(gòu)對(duì)粒徑和細(xì)胞轉(zhuǎn)染效率的關(guān)系，建立了相應(yīng)的構(gòu)效關(guān)系。結(jié)果表明通過(guò)采用不同比例的PEI，得到不同接枝量對(duì)結(jié)構(gòu)、粒徑和穩(wěn)定性的影響。當(dāng)接枝量小時(shí)，粒徑較大，PEI與PEI之間的作用力小，納米粒子為舒展的狀態(tài)，并形成了插層的結(jié)構(gòu)，相對(duì)應(yīng)的穩(wěn)定性則下降；當(dāng)接枝量逐漸增加粒徑卻逐漸變小，PEI與PEI之間的作用力增大，納米粒子為卷曲的狀態(tài)，形成了剝離的結(jié)構(gòu)，，穩(wěn)定性也由于作用力的作用增強(qiáng)；當(dāng)接枝量逐漸增加的緩慢，粒徑也幾乎趨于平穩(wěn)，作用力之間達(dá)到恒定，穩(wěn)定性達(dá)到最好。我們制備了結(jié)構(gòu)可控，粒徑小，穩(wěn)定性好的基因載體。同時(shí)GO-PEI-PEG-FA/si-Stat3復(fù)合物沉默了Stat3的表達(dá)使達(dá)到了靶向治療肝癌的目的。
[Abstract]:Gene therapy is a hot spot in the treatment of diseases, especially cancer. The key problem of gene therapy is to develop a suitable and efficient gene vector, which can be divided into viral vector and non-viral vector. Viral carriers are less used because of their high toxicity and limitations. Non-viral carriers have been widely used because of their high safety and effectiveness, such as polymers, liposomes, nanoparticles and so on. In recent years, although non-viral gene vectors have made great progress, some problems, such as non-specific targeting sites, toxicity and biological stability, have not been solved. Therefore, the development of an effective nano-carrier remains the biggest challenge in gene therapy. Graphene, which has a two-dimensional layered structure with thick carbon atoms, has been widely used in many fields, such as electrochemical devices, biological applications and so on. Graphene oxide (GOO) is a water-soluble derivative of graphene. Because of its easy preparation, good dispersion in aqueous solution and physiological environment, good colloid stability has attracted great interest, and has attracted great interest in biosensors. Gene and drug delivery, as well as biological fields such as imaging in vivo and in vitro, have also shown potential applications. In this paper, based on graphene oxide, a novel multifunctional graphene oxide gene vector GO-PEI-PEG-FA-FA-Stat3 was designed by chemical coupling modification. In order to improve the transfection efficiency, we selected the commonly used polymer branched polyethyleneimine (PEI) and carboxylated graphene oxide by azo chemical reaction. In order to improve the biological stability of the solution, a small molecule of folate, FAA, was introduced and modified to achieve the purpose of targeted therapy. The GO-PEI-PEG-FA was connected with si-Stat3 plasmid by electrostatic interaction. Finally, the GO-PEI-PEG-FA/si-Stat3 gene vector was formed. We studied the effect of different proportion of PEI on the structure of GO-PEI-PEG-FA, and further studied the relationship of structure to particle size and cell transfection efficiency during this process, and established the corresponding structure-activity relationship. The results show that the effects of grafting amount on structure, particle size and stability are obtained by using different proportion of PEI. When the grafting amount is small, the interaction force between the larger particle size and the PEI is small, and the nanoparticles are in a stretch state, and the intercalation structure is formed, and the corresponding stability decreases. When the grafted amount increases gradually but the interaction force between PEI and PEI increases gradually, the nanoparticles are crimped, and the structure of peeling is formed, and the stability is enhanced because of the effect of the acting force, and when the grafting amount increases slowly, The particle size is almost stable, the interaction force is constant, and the stability is the best. We have prepared gene vectors with controllable structure, small particle size and good stability. At the same time, the GO-PEI-PEG-FA/si-Stat3 complex silenced the expression of Stat3, so that the target therapy of liver cancer was achieved.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R450;O613.71

【共引文獻(xiàn)】

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