本文選題：納米氧化鋅 + HEK293細(xì)胞　； 參考：《蘇州大學(xué)》2015年碩士論文

【摘要】：納米氧化鋅(Zn O Nanoparticles,Zn O NPs)是目前繼碳納米管之后又一個備受關(guān)注的多功能納米材料。納米氧化鋅的顆粒大小約在1~100nm之間。由于晶粒的細(xì)微化,其表面電子結(jié)構(gòu)和晶體結(jié)構(gòu)發(fā)生變化,產(chǎn)生了宏觀物體所不具有的表面效應(yīng)、體積效應(yīng)、量子尺寸效應(yīng)和宏觀隧道效應(yīng)以及高透明度、高分散性等特點,而被廣泛應(yīng)用于橡膠、陶瓷、化工、環(huán)保、飼料、電子、化妝品、光學(xué)、生物、醫(yī)藥等眾多領(lǐng)域,具有非常廣闊的應(yīng)用前景。納米氧化鋅在創(chuàng)造巨大經(jīng)濟(jì)效益的同時,其毒性和健康效應(yīng)也應(yīng)引起人們的關(guān)注。但是,到目前為止,關(guān)于納米氧化鋅的生物安全性的研究還很少,而且已取得的研究結(jié)果也并不統(tǒng)一,還需要進(jìn)行大量的研究證明。基于此,本研究以三種不同尺寸的納米氧化鋅(15nm、30nm、90nm)為對象,比較不同濃度的納米氧化鋅(15nm)對HEK293細(xì)胞的毒性大小以及同一濃度(4.0μg/ml)不同尺寸的氧化鋅(常規(guī)氧化鋅、15nm、30nm、90nm)對HEK293細(xì)胞的毒性差異,并進(jìn)一步研究納米氧化鋅的毒性作用機(jī)制,以期為納米氧化鋅做體外安全性評估,并為納米材料毒理學(xué)以及其生物安全性研究提供一定的理論依據(jù)。首先,利用透射電子顯微鏡(TEM)和動態(tài)光散射(DLS)對實驗所用納米氧化鋅(15nm、30nm、90nm)進(jìn)行表征,然后分別從納米氧化鋅對HEK293細(xì)胞的細(xì)胞形態(tài)學(xué)、細(xì)胞損傷、抗氧化指標(biāo)等方面的影響進(jìn)行研究。研究結(jié)果顯示,實驗所用3種不同尺寸Zn O NPs的純度均大于99.9%,其結(jié)構(gòu)均為紅鋅礦晶型,其尺寸均比理論尺寸略大。用15nm不同濃度(0、0.5、1.0、2.0、4.0、8.0、16.0μg/ml)的Zn O NPs、同一濃度(4.0μg/ml)不同尺寸(常規(guī)Zn O、15nm、30nm、90nm)的Zn O NPs處理HEK293細(xì)胞,培養(yǎng)48h后,熒光顯微鏡觀察顯示,隨著Zn O NPs濃度的逐漸增大,HEK293細(xì)胞逐漸出現(xiàn)形態(tài)學(xué)變化:細(xì)胞排列疏松,間隙增大,胞質(zhì)內(nèi)顆粒物增多,透明度下降,部分細(xì)胞回縮變圓并出現(xiàn)脫落。且在相同濃度下,Zn O NPs尺寸越小,這種變化越明顯。MTT、流式細(xì)胞儀、Caspase3檢測結(jié)果顯示,隨著Zn O NPs濃度的增大,HEK293細(xì)胞活性逐漸降低,凋亡率逐漸升高,Caspase3活性逐漸增強(qiáng);隨著Zn O NPs尺寸的減小,HEK293細(xì)胞活性逐漸降低,凋亡率逐漸升高,Caspase3活性逐漸增強(qiáng)。HEK293細(xì)胞抗氧化指標(biāo)(MDA、ROS、GSH)檢測結(jié)果顯示,隨著Zn O NPs濃度的增大,HEK293細(xì)胞中MDA和ROS水平逐漸升高,GSH水平逐漸降低;隨著Zn O NPs尺寸的減小,HEK293細(xì)胞中MDA和ROS水平逐漸升高,GSH水平逐漸降低。MDA、ROS的過量積累,GSH的大量消耗,影響了細(xì)胞正常的氧化-抗氧化水平,誘導(dǎo)HEK293細(xì)胞出現(xiàn)氧化應(yīng)激狀態(tài),從而進(jìn)一步誘導(dǎo)Caspase3的活化,最后導(dǎo)致細(xì)胞發(fā)生凋亡,且Zn O NPs對HEK293細(xì)胞的這種毒性作用具有明顯的劑量效應(yīng)和尺寸效應(yīng),即Zn O NPs濃度越大,其對HEK293細(xì)胞的毒性越大,Zn O NPs尺寸越小,其對HEK293細(xì)胞的毒性越大。
[Abstract]:Nano-ZnO Zn-O Nanoparticles-Zn-O NPsis another multifunctional nano-material, which has attracted much attention after carbon nanotubes (CNTs). The particle size of nanometer zinc oxide is about between 1~100nm. Due to the fineness of the grain, the surface electronic structure and crystal structure change, resulting in the surface effect, volume effect, quantum size effect, macroscopic tunnel effect, high transparency, high dispersion and so on. It is widely used in many fields, such as rubber, ceramics, chemical industry, environmental protection, feed, electronics, cosmetics, optics, biology, medicine and so on. At the same time, the toxicity and health effect of nanometer zinc oxide should be paid more attention to. However, so far, there are few studies on the biosafety of nano-zinc oxide, and the results obtained are not uniform, and a lot of research is needed to prove it. Based on this, three kinds of nanometer zinc oxide with different sizes were studied. To compare the toxicity of different concentrations of nanometer zinc oxide (15 nm) to HEK293 cells and the toxicity of different sizes of zinc oxide (conventional zinc oxide 15 nm / ml) to HEK293 cells, and to further study the mechanism of the toxicity of nanometer zinc oxide. In order to do in vitro safety evaluation of nano-zinc oxide, and provide a theoretical basis for the toxicology of nano-materials and their biological safety research. First, transmission electron microscopy (TEM) and dynamic light scattering (DLSs) were used to characterize the HEK293 cells. The effects of antioxidant indexes were studied. The results show that the purity of three different sizes of Zno NPs is greater than 99.9, and the structure of all of them is red zinc ore crystal type, which is slightly larger than the theoretical size. HEK293 cells were treated with Zn O NPs with different concentrations of Zn O NPs, of different concentration (15nm) of 0.5nmnmnmnmnmnmnmnm ~ (30 nm) and 4.0 渭 g / ml of 4.0 渭 g / ml of Zn O NPs, (4.0 渭 g / ml). The cells were cultured for 48 h, and the results of fluorescence microscopy showed that the HEK293 cells were cultured for 48 hours, and the results were as follows: (1) after 48 h culture, the cells were treated with Zn O NPs of the same concentration of 4.0 渭 g / ml. With the increasing of Zn O NPs concentration, morphological changes of HEK293 cells appeared gradually: the arrangement of cells was loose, the gap was enlarged, the particles in the cytoplasm were increased, the transparency was decreased, and some cells were reduced to round and shedding. The smaller the size of Zn-O NPs at the same concentration, the more obvious the change was. The results of flow cytometry showed that the activity of HEK293 cells decreased with the increase of Zn O NPs concentration, and the apoptosis rate increased gradually. With the decrease of Zno NPs size, the activity of HEK293 cells decreased, the apoptosis rate increased and the activity of Caspase3 increased. The results showed that the antioxidant index of HEK293 cells was determined by MDA-ROSS-GSH. With the increase of Zn O NPs concentration, the levels of MDA and ROS in HEK293 cells gradually increased, and the levels of MDA and ROS in HEK293 cells gradually decreased with the decrease of Zn O NPs size. It affected the normal level of oxidation-antioxidation, induced oxidative stress in HEK293 cells, further induced the activation of Caspase3, and finally resulted in cell apoptosis. The toxicity of Zn O NPs to HEK293 cells showed significant dose and size effects, that is, the greater the concentration of Zn O NPs, the greater the size of Zn O NPs and the greater the toxicity to HEK293 cells.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：TB383.1;X171.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 田靜博;劉琳;錢建華;邢錦娟;秦金蘭;;納米氧化鋅的制備技術(shù)與應(yīng)用研究進(jìn)展[J];化學(xué)工業(yè)與工程技術(shù);2008年02期

2 汪冰,豐偉悅,趙宇亮,邢更妹,柴之芳,王海芳,賈光;納米材料生物效應(yīng)及其毒理學(xué)研究進(jìn)展[J];中國科學(xué)(B輯 化學(xué));2005年01期

3 劉曉慧;郭利利;秦定霞;高莉;楊明德;張紅梅;崔毓桂;劉嘉茵;;ZnO納米對雌性ICR小鼠急性毒性作用的研究[J];南京醫(yī)科大學(xué)學(xué)報(自然科學(xué)版);2009年02期

4 李巖;彭光銀;何胡軍;喬永康;常青;楊旭;;碳納米管導(dǎo)致小鼠肺部急性氧化損傷作用的研究[J];生態(tài)毒理學(xué)報;2006年04期

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