本文選題：原子力顯微鏡 + 單分子力示蹤�。� 參考：《長春工業(yè)大學(xué)》2017年碩士論文

【摘要】：相比于眾多傳統(tǒng)的高分子藥物載體,樹枝狀大分子聚合物聚酰胺胺(PAMAM)具有很多特有的優(yōu)勢,它的表面連有大量的官能團,經(jīng)過修飾可以連接更多的藥物分子;其次,在空間上呈球狀分布的高代數(shù)樹枝狀PAMAM大分子,內(nèi)部存在著較大的孔腔,這些孔腔可以包埋藥物分子。因此越來越多的研究人員著手研究PAMAM作為藥物載體的應(yīng)用,然而由于研究手段的空間和時間分辨率限制,作為藥物載體進入細胞的第一步——跨膜過程并不是很清楚。在此,我們利用基于原子力顯微鏡(AFM)的單分子力示蹤技術(shù)(可以測量皮牛級的力和微秒量級的動態(tài)過程)對單個第五代樹枝狀化合物分子(G5-PAMAM)進入細胞的跨膜動態(tài)過程進行研究。我們實時記錄了單個PAMAM進入活細胞的動態(tài)過程,測量到跨膜力的大小及其進入細胞所需的時間,并研究了其進入細胞的內(nèi)吞機制。本研究將為PAMAM作為藥物載體的應(yīng)用提供更多的理論基礎(chǔ),對生物醫(yī)學(xué)領(lǐng)域有重要意義。研究取得進展如下:1.本研究記錄了單個G5-PAMAM進入活細胞的動態(tài)過程,測量到跨膜力的大小及其進入細胞所需要的時間。G5-PAMAM納米粒子與細胞作用的內(nèi)吞力的范圍是50pN至425 pN,其平均的作用力是172±74 pN,進入細胞內(nèi)所需時間的范圍是2.5 ms至65 ms,平均值為19±11 ms。2.另外,本研究還揭示了G5-PAMAM進入細胞的內(nèi)吞機制。不僅用力示蹤技術(shù)在單分子水平上做了對照和阻礙實驗,而且也通過激光共聚焦顯微鏡量化了相對于對照實驗(未用阻礙試劑處理)而言用阻礙試劑預(yù)處理后的細胞內(nèi)G5-PAMAM粒子的平均熒光強度。值得注意的是,兩種實驗方法得出的結(jié)論一致。非洲綠猴腎細胞(Vero)內(nèi)吞G5-PAMAM通過小窩蛋白和網(wǎng)格蛋白介導(dǎo)的內(nèi)吞及巨胞飲方式(網(wǎng)格蛋白及巨胞飲為主)完成,其中網(wǎng)格蛋白介導(dǎo)和巨胞飲是主要的途徑。
[Abstract]:Compared with many traditional polymer drug carriers, the dendritic macromolecular polymer PAMAM has many unique advantages. It has a large number of functional groups attached to its surface, which can be modified to connect more drug molecules. High algebraic dendritic PAMAM macromolecules with spherical distribution in space have large pore cavities which can be encapsulated into drug molecules. Therefore, more and more researchers have begun to study the application of PAMAM as drug carrier. However, due to the limitation of space and time resolution, the first step-transmembrane process of drug carrier entering cells is not very clear. In this paper, the transmembrane dynamic process of single fifth generation dendrimer (G5-PAMAM) into cells was studied by using AFM-based single molecular force tracer technique (which can measure the dynamic processes of skin cattle force and microsecond order of magnitude). We recorded the dynamic process of a single PAMAM entering living cells in real time, measured the transmembrane force and the time it took to enter the cells, and studied the endocytosis mechanism. This study will provide more theoretical basis for the application of PAMAM as a drug carrier, and will be of great significance to the biomedical field. The progress of the study is as follows: 1. In this study, the dynamic process of single G5-PAMAM entering living cells was recorded. The transmembrane force and the time required to enter the cell. The endocytosis range of G5-PAMAM nanoparticles was from 50pN to 425pN, the average force was 172 鹵74 PN, and the time to enter the cell was 2.5 Ms to 65 Ms, with an average of 19 鹵11 ms.2. In addition, this study also revealed the endocytosis mechanism of G5-PAMAM entering cells. Not only were forced tracer techniques used to control and block experiments at the single molecular level, The average fluorescence intensity of G5-PAMAM particles pretreated with hindrance reagent was quantified by laser confocal microscopy. It is worth noting that the two experimental methods come to the same conclusion. The endocytosis of G5-PAMAM in the kidney cells of African green monkey is accomplished by the way of endocytosis and giant cell drink (mainly grid protein and giant cell drink) mediated by nest protein and grid protein, among which grid protein and giant cell drink are the main pathway.
【學(xué)位授予單位】：長春工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：TQ460.1;O633.22

【參考文獻】

相關(guān)期刊論文 前3條

1 Felipe Vidal;Leonardo Guzman;;Dendrimer nanocarriers drug action: perspective for neuronal pharmacology[J];Neural Regeneration Research;2015年07期

2 韓巧榮;王炳祥;丁馬太;何旭敏;;Cu~(2+)對樹狀大分子PAMAM-FCD熒光性能的影響[J];高等學(xué)�；瘜W(xué)學(xué)報;2010年01期

3 潘碧峰;崔大祥;徐萍;陳浩;劉鳳濤;李清;黃拓;尤曉剛;邵君;鮑晨晨;高峰;賀蓉;舒孟軍;馬勇杰;;Design of Dendrimer Modified Carbon Nanotubes for Gene Delivery[J];Chinese Journal of Cancer Research;2007年01期

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本文編號：1903100