【摘要】：研究目的： 1、通過(guò)研究硫化鋅包被硒化鎘量子點(diǎn)（CdSe/ZnS QDs）基本物理及光學(xué)特征，驗(yàn)證其是否為一種適用于生物醫(yī)學(xué)的熒光染料。 2、通過(guò)檢測(cè)CdSe/ZnS QDs及氯化鎘（CdCl2）對(duì)Hela細(xì)胞活性及凋亡率的影響，探究CdSe/ZnS QDs是否具有細(xì)胞毒性作用。 3、通過(guò)比較空白對(duì)照組、CdSe/ZnS QDs組及CdCl2組Hela細(xì)胞內(nèi)ROS產(chǎn)量，探究CdSe/ZnS QDs誘導(dǎo)Hela細(xì)胞凋亡可能的機(jī)制。 4、通過(guò)對(duì)凋亡相關(guān)蛋白的檢測(cè)，進(jìn)一步闡述CdSe/ZnS QDs誘導(dǎo)細(xì)胞凋亡的機(jī)制。 方法： 本實(shí)驗(yàn)采用透射電子顯微鏡（Transmission electron microscope，TEM）、紫外分光光度計(jì)（Ultraviolet spectrophotometer）及熒光光譜儀（Fluorescencespectrometer）對(duì)CdSe/ZnS QDs進(jìn)行表征。透射電子顯微鏡用于了解CdSe/ZnSQDs的大小、形狀、外觀均一程度，紫外分光光度計(jì)用于檢測(cè)CdSe/ZnS QDs的吸收光波長(zhǎng)范圍，熒光光度計(jì)用于檢測(cè)CdSe/ZnS QDs所發(fā)射熒光波長(zhǎng)范圍。 在建立穩(wěn)定Hela傳代細(xì)胞株，，通過(guò)MTT實(shí)驗(yàn)比較不同濃度CdSe/ZnS QDs及CdCl2孵育Hela細(xì)胞24h后細(xì)胞活性的改變，再通過(guò)流式細(xì)胞術(shù)比較不同濃度CdSe/ZnS QDs及CdCl2對(duì)Hela細(xì)胞凋亡率的影響。通過(guò)對(duì)比空白對(duì)照組、CdSe/ZnS QDs組及CdCl2組Hela細(xì)胞活性及凋亡率差異，初步判斷CdSeCdSe/ZnS QDs是否具有細(xì)胞毒性。 通過(guò)DCFH-DA熒光探針檢測(cè)不同濃度CdSe/ZnS QDs及CdCl2孵育Hela細(xì)胞24h后細(xì)胞內(nèi)ROS量，并比較不同組之間ROS量的差異，探究CdSe/ZnSQDs對(duì)Hela細(xì)胞內(nèi)ROS量的影響。 最后通過(guò)Western blot檢測(cè)不同濃度CdSe/ZnS QDs及CdCl2孵育Hela細(xì)胞24h后細(xì)胞內(nèi)凋亡相關(guān)蛋白Bcl-2與Bax表達(dá)量，對(duì)比不同組間Bcl-2與Bax蛋白表達(dá)量差異分析CdSe/ZnS QDs對(duì)細(xì)胞內(nèi)Bcl-2與Bax蛋白表達(dá)的影響并分析其可能的原因。 結(jié)果： 1、本實(shí)驗(yàn)所用CdSe/ZnS QDs顆粒均勻呈球形，直徑小，具有良好的單分散性和晶體結(jié)構(gòu)，激發(fā)光譜寬，發(fā)射光譜連續(xù)且對(duì)稱。 2、CdSe/ZnS QDs對(duì)Hela細(xì)胞的毒性作用遠(yuǎn)強(qiáng)于CdCl2，且其細(xì)胞毒性作用與給藥濃度呈正比。 3、 CdSe/ZnS QDs較CdCl2誘導(dǎo)Hela內(nèi)產(chǎn)生更多的ROS，且CdSeQDs/ZnS對(duì)ROS的誘導(dǎo)效應(yīng)與給藥濃度成正比。 4、CdSe/ZnS QDs較CdCl2誘導(dǎo)Hela細(xì)胞凋亡效應(yīng)更強(qiáng)，且其誘導(dǎo)凋亡效應(yīng)與給藥濃度成正比。 5、CdSe/ZnS QDs處理組細(xì)胞內(nèi)Bcl-2蛋白表達(dá)量較空白對(duì)照組及相應(yīng)CdCl2處理組少，而Bax蛋白表達(dá)較空白對(duì)照組及相應(yīng)CdCl2處理組多。 結(jié)論： 1、CdSe/ZnS QDs直徑小，光學(xué)特性優(yōu)，適用于生物醫(yī)學(xué)研究。 2、CdSe/ZnS QDs具有細(xì)胞毒性作用，其毒性作用強(qiáng)于CdCl2，且毒性作用與給藥濃度成正比。 3、CdSe/ZnS QDs能通過(guò)促進(jìn)細(xì)胞內(nèi)ROS生成而誘導(dǎo)Hela細(xì)胞凋亡，且CdSe/ZnS QDs能通過(guò)誘導(dǎo)Bax表達(dá)增加、Bcl-2表達(dá)減少而誘導(dǎo)Hela細(xì)胞凋亡。
[Abstract]:Objective: 1. By studying the basic physical and optical properties of zinc sulfide coated cadmium selenide quantum dots (CdSe/ZnS QDs), it is proved that it is a kind of fluorescent dye suitable for biomedicine. 2. By examining the effects of CdSe/ZnS QDs and cadmium chloride (CdCl2) on the activity and apoptosis rate of Hela cells, we investigated the cytotoxicity of CdSe/ZnS QDs. 3. By comparing ROS production in Hela cells of blank control group, CdSe/ZnS QDs group and CdCl2 group, the possible mechanism of Hela cell apoptosis induced by CdSe/ZnS QDs was explored. 4. Through the detection of apoptosis-related proteins, the mechanism of apoptosis induced by CdSe/ZnS QDs was further explained. Methods: CdSe/ZnS QDs was characterized by transmission electron microscope (Transmission electron microscope,TEM), ultraviolet spectrophotometer (Ultraviolet spectrophotometer) and fluorescence spectrometer (Fluorescencespectrometer). Transmission electron microscopy (TEM) is used to understand the size, shape and appearance of CdSe/ZnSQDs. Ultraviolet spectrophotometer is used to detect the absorption wavelength range of CdSe/ZnS QDs, and fluorescence photometer is used to detect the range of fluorescence wavelength emitted by CdSe/ZnS QDs. After establishing stable Hela cell lines, the changes of cell activity of Hela cells incubated with different concentrations of CdSe/ZnS QDs and CdCl2 for 24 hours were compared by MTT assay, and the effects of different concentrations of CdSe/ZnS QDs and CdCl2 on the apoptosis rate of Hela cells were compared by flow cytometry. By comparing the difference of Hela cell activity and apoptosis rate among control group, CdSe/ZnS QDs group and CdCl2 group, the cytotoxicity of CdSeCdSe/ZnS QDs was preliminarily determined. The amount of ROS in Hela cells incubated with different concentrations of CdSe/ZnS QDs and CdCl2 for 24 hours was detected by DCFH-DA fluorescence probe, and the difference of ROS content among different groups was compared to explore the effect of CdSe/ZnSQDs on the quantity of ROS in Hela cells. Finally, Western blot was used to detect the expression of Bcl-2 and Bax in Hela cells incubated with different concentrations of CdSe/ZnS QDs and CdCl2 for 24 hours. The effect of CdSe/ZnS QDs on the expression of Bcl-2 and Bax in different groups was analyzed and the possible causes were analyzed. Results: 1. The CdSe/ZnS QDs particles used in this experiment are spherical and small in diameter, with good monodispersity and crystal structure, wide excitation spectrum, continuous and symmetrical emission spectrum. 2 the cytotoxic effect of CDS / ZnS QDs on Hela cells was much stronger than that on CdCl2, cells, and its cytotoxicity was proportional to the drug concentration. 3. More ROS, was produced in Hela induced by CdSe/ZnS QDs than that induced by CdCl2, and the effect of CdSeQDs/ZnS on ROS was directly proportional to the drug concentration. 4CdSee / ZnS QDs was more effective than CdCl2 in inducing apoptosis of Hela cells, and the apoptotic effect was in direct proportion to the drug concentration. 5The expression of Bcl-2 protein in CDSe-ZnS QDs treated group was lower than that in control group and CdCl2 treatment group, but the expression of Bax protein was higher than that in blank control group and CdCl2 treatment group. Conclusion: 1 CdSeR / ZnS QDs has small diameter and excellent optical properties, which is suitable for biomedical research. 2CdSe- / ZnS QDs has cytotoxic effect, which is stronger than CdCl2, and is directly proportional to the concentration of the drug. 3CdSee / ZnS QDs could induce apoptosis of Hela cells by promoting the production of intracellular ROS, and CdSe/ZnS QDs could induce apoptosis of Hela cells by increasing the expression of Bax and decreasing the expression of Bcl-2.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.3

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