巰基穩(wěn)定的CdTe量子點(diǎn)與蛋白質(zhì)相互作用及其細(xì)胞毒性的研究
發(fā)布時(shí)間：2018-08-20 14:53
【摘要】：1.概述了量子點(diǎn)的基本物理特性、有機(jī)相和水相合成方法、表面修飾方法,及其在分析科學(xué)和生物醫(yī)學(xué)領(lǐng)域的應(yīng)用及毒理效應(yīng)的研究現(xiàn)狀與進(jìn)展。在此基礎(chǔ)上,提出了本文的研究構(gòu)想。 2.采用水熱合成法,分別以N-乙�；�-L-半胱氨酸(NAC)和巰基丙酸(MPA)為穩(wěn)定劑,制備出不同粒徑的高質(zhì)量碲化鎘(CdTe)量子點(diǎn)。利用紫外可見吸收光譜、熒光光譜、傅立葉變換紅外光譜和透射電鏡等手段對(duì)所制備的NAC/MPA-CdTe QDs進(jìn)行了表征。結(jié)果表明,所制備的量子點(diǎn)粒徑分布均一,紫外吸收譜寬而連續(xù),熒光發(fā)射峰窄且對(duì)稱,發(fā)射范圍幾乎覆蓋了整個(gè)可見區(qū)(480-760nm)。通過改變反應(yīng)時(shí)間實(shí)現(xiàn)了對(duì)量子點(diǎn)熒光發(fā)射波長(zhǎng)的調(diào)制。 3.主要利用熒光光譜法,分別研究了生理?xiàng)l件下CdTe量子點(diǎn),CdTe/CdS核-殼量子點(diǎn)與細(xì)胞色素c(cytochrome c, Cyt c),以及CdTe量子點(diǎn)與牛血清白蛋白(bovine serum albumin, BSA)之間的相互作用。研究發(fā)現(xiàn)Cytc對(duì)不同粒徑的量子點(diǎn)均有顯著的熒光猝滅作用；BSA的引入則使量子點(diǎn)的熒光明顯增強(qiáng),而量子點(diǎn)對(duì)BSA的熒光表現(xiàn)出明顯猝滅效應(yīng)。探討了量子點(diǎn)對(duì)牛血清白蛋白的熒光猝滅效應(yīng)、牛血清白蛋白對(duì)量子點(diǎn)的熒光增敏效應(yīng),以及細(xì)胞色素c對(duì)量子點(diǎn)熒光猝滅效應(yīng)的作用機(jī)制,為進(jìn)一步研究量子點(diǎn)與活細(xì)胞內(nèi)蛋白質(zhì)之間的相互作用提供了實(shí)驗(yàn)依據(jù)。 4.利用激光共聚焦顯微鏡分別研究了水溶性MPA-CdTe QD和NAC-CdTe QD與小鼠表皮黑色素細(xì)胞B16-F10、人神經(jīng)膠質(zhì)瘤細(xì)胞SH-SY5Y的相互作用。初步結(jié)果顯示,相同條件下,MPA-CdTe QD進(jìn)入細(xì)胞的能力強(qiáng)于NAC-CdTe QD; QD進(jìn)入細(xì)胞的能力存在細(xì)胞種別依賴性；外源性物種對(duì)QD在細(xì)胞內(nèi)的分布存在一定的協(xié)同效應(yīng)。為了進(jìn)一步提高量子點(diǎn)的生物相容性,利用化學(xué)還原的BSA對(duì)水溶性CdTeQDs進(jìn)行了表而修飾。由紫外光譜,熒光光譜和細(xì)胞存活率檢測(cè)實(shí)驗(yàn)結(jié)果表明,變性BSA被連接到CdTe QDs表面,提高了量子點(diǎn)的化學(xué)穩(wěn)定性和生物相容性。此外,對(duì)不同穩(wěn)定劑(MPA, NAC和dBSA)修飾的量子點(diǎn)與細(xì)胞之間的相互作用進(jìn)行了初步比較研究,結(jié)果表明,在相同濃度下MPA-CdTe QDs對(duì)細(xì)胞存活率影響最大,NAC-CdTe QDs對(duì)細(xì)胞存活率的影響次之,dBSA-CdTe QDs對(duì)細(xì)胞存活率的影響最小。實(shí)驗(yàn)還表明紫外光照射等外部因素會(huì)增強(qiáng)量子點(diǎn)的細(xì)胞毒性。上述研究為合成高生物相容性的量子點(diǎn),最大限度地抑制其細(xì)胞毒性以滿足生物醫(yī)學(xué)方面應(yīng)用的要求提供了一定的理論依據(jù)。
[Abstract]:1. The basic physical properties of quantum dots, organic and aqueous phase synthesis methods, surface modification methods, their applications in analytical science and biomedicine, and the research status and progress of toxicological effects are reviewed in this paper. On the basis of this, the research conception of this paper is put forward. 2. Using N-acetyl-L-cysteine (NAC) and mercaptopropionic acid (MPA) as stabilizers, high-quality cadmium telluride (CdTe) quantum dots with different particle sizes were prepared by hydrothermal synthesis. The NAC/MPA-CdTe QDs was characterized by UV-Vis absorption spectrum, fluorescence spectrum, Fourier transform infrared spectroscopy and transmission electron microscope. The results show that the size distribution of the QDs is uniform, the UV absorption spectrum is wide and continuous, the fluorescence emission peak is narrow and symmetrical, and the emission range almost covers the whole visible region (480-760nm). The fluorescence emission wavelength of quantum dots was modulated by changing the reaction time. 3. 3. The interaction of CdTe quantum dots with cytochrome c (cytochrome c, Cyt c), and CdTe quantum dots with bovine serum albumin (BSA) under physiological conditions was studied by fluorescence spectroscopy. It is found that Cytc has a significant fluorescence quenching effect on quantum dots with different particle sizes. The introduction of Cytc enhances the fluorescence of quantum dots, while the fluorescence of BSA is obviously quenched by quantum dots. The fluorescence quenching effect of quantum dot on bovine serum albumin, the fluorescence enhancement effect of bovine serum albumin on quantum dot and the mechanism of fluorescence quenching effect of cytochrome c on quantum dot were studied. It provides an experimental basis for the further study of the interaction between quantum dots and proteins in living cells. 4. The interaction of water-soluble MPA-CdTe QD and NAC-CdTe QD with mouse epidermal melanocyte B16-F10 and human glioma cell SH-SY5Y was studied by laser confocal microscopy. The preliminary results showed that the ability of MPA-CdTe QD to enter the cells was stronger than that of NAC-CdTe QD under the same conditions, and the ability of QD to enter the cells was species dependent, and there was a synergistic effect of exogenous species on the distribution of QD in the cells. In order to further improve the biocompatibility of quantum dots, chemically reduced BSA was used to modify water-soluble CdTeQDs. The results of UV spectra, fluorescence spectra and cell survival test show that denatured BSA is connected to the surface of CdTe QDs, which improves the chemical stability and biocompatibility of QDs. In addition, the interaction between QDs modified by different stabilizers, (MPA, NAC and dBSA, was studied. At the same concentration, MPA-CdTe QDs had the greatest effect on cell survival. NAC-CdTe QDs had the second best effect on cell survival, and dBSA-CdTe QDs had the least effect on cell survival. The experiments also show that external factors such as ultraviolet irradiation can enhance the cytotoxicity of quantum dots. These studies provide a theoretical basis for the synthesis of highly biocompatible quantum dots and the maximum inhibition of their cytotoxicity to meet the requirements of biomedical applications.
【學(xué)位授予單位】：山西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R114
本文編號(hào)：2194040
[Abstract]:1. The basic physical properties of quantum dots, organic and aqueous phase synthesis methods, surface modification methods, their applications in analytical science and biomedicine, and the research status and progress of toxicological effects are reviewed in this paper. On the basis of this, the research conception of this paper is put forward. 2. Using N-acetyl-L-cysteine (NAC) and mercaptopropionic acid (MPA) as stabilizers, high-quality cadmium telluride (CdTe) quantum dots with different particle sizes were prepared by hydrothermal synthesis. The NAC/MPA-CdTe QDs was characterized by UV-Vis absorption spectrum, fluorescence spectrum, Fourier transform infrared spectroscopy and transmission electron microscope. The results show that the size distribution of the QDs is uniform, the UV absorption spectrum is wide and continuous, the fluorescence emission peak is narrow and symmetrical, and the emission range almost covers the whole visible region (480-760nm). The fluorescence emission wavelength of quantum dots was modulated by changing the reaction time. 3. 3. The interaction of CdTe quantum dots with cytochrome c (cytochrome c, Cyt c), and CdTe quantum dots with bovine serum albumin (BSA) under physiological conditions was studied by fluorescence spectroscopy. It is found that Cytc has a significant fluorescence quenching effect on quantum dots with different particle sizes. The introduction of Cytc enhances the fluorescence of quantum dots, while the fluorescence of BSA is obviously quenched by quantum dots. The fluorescence quenching effect of quantum dot on bovine serum albumin, the fluorescence enhancement effect of bovine serum albumin on quantum dot and the mechanism of fluorescence quenching effect of cytochrome c on quantum dot were studied. It provides an experimental basis for the further study of the interaction between quantum dots and proteins in living cells. 4. The interaction of water-soluble MPA-CdTe QD and NAC-CdTe QD with mouse epidermal melanocyte B16-F10 and human glioma cell SH-SY5Y was studied by laser confocal microscopy. The preliminary results showed that the ability of MPA-CdTe QD to enter the cells was stronger than that of NAC-CdTe QD under the same conditions, and the ability of QD to enter the cells was species dependent, and there was a synergistic effect of exogenous species on the distribution of QD in the cells. In order to further improve the biocompatibility of quantum dots, chemically reduced BSA was used to modify water-soluble CdTeQDs. The results of UV spectra, fluorescence spectra and cell survival test show that denatured BSA is connected to the surface of CdTe QDs, which improves the chemical stability and biocompatibility of QDs. In addition, the interaction between QDs modified by different stabilizers, (MPA, NAC and dBSA, was studied. At the same concentration, MPA-CdTe QDs had the greatest effect on cell survival. NAC-CdTe QDs had the second best effect on cell survival, and dBSA-CdTe QDs had the least effect on cell survival. The experiments also show that external factors such as ultraviolet irradiation can enhance the cytotoxicity of quantum dots. These studies provide a theoretical basis for the synthesis of highly biocompatible quantum dots and the maximum inhibition of their cytotoxicity to meet the requirements of biomedical applications.
【學(xué)位授予單位】：山西大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R114
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