纳米二氧化硅颗粒对人血管内皮细胞损伤作用剂量和时间效应的研究

发布时间：2018-05-13 20:31

本文选题：纳米二氧化硅颗粒 + 血管内皮细胞　；参考：《华中科技大学》2013年硕士论文

【摘要】：目的:纳米材料的职业安全和健康问题是很复杂的。纳米材料虽然粒径小却有较大的表面积，导致其与相同成分较大尺寸的颗粒物相比，物理、化学、生物学性质发生较大的变化，这些理化性质极有可能会影响纳米材料以下几方面的能力，到达肺气体交换区域、自肺进入运输至整个机体、穿透皮肤屏障、跨越细胞膜，甚至在分子水平的反应等等。初步的一些研究结果表明，微米级物质的安全性评价结果可能并不不适用于纳米级物质，因此纳米尺度材料的生物安全性问题得到人们的广泛关注。故本研究选择比较常见的典型纳米颗粒（SiO2颗粒）作为染尘颗粒，通过体外实验对纳米颗粒物心血管毒性的剂量和时间效应进行研究，为纳米级化学物的心血管安全性评价提供进一步的实验依据。方法:选择nano-SiO2作为染尘颗粒物，将人脐静脉内皮细胞（HUVECs）作为受试细胞，不同剂量（0、5、10、25、50、100、200μg/ml）的颗粒物和人脐静脉内皮细胞共培养不同时间6、12、24、48h，用CCK-8方法检测细胞存活率，计算半数致死量，确定后续实验的染尘剂量为0、5、10、20、40μg/ml。将上述剂量（0、5、10、20、40μg/ml）的nano-SiO2颗粒与HUVECs共培养6、12、24、48h。用相应试剂盒测定细胞培养液上清中乳酸脱氢酶（LDH）活性、总超氧化物歧化酶（SOD）活性、肿瘤坏死因子-α（TNF-α）释放量、白介素-6（IL-6）的释放量。结果:细胞存活率随着nano-SiO2染尘剂量升高和作用时间的延长而逐渐降低。如果作用时间短，细胞存活率在较高剂量开始明显降低；若作用时间长，细胞存活率在低剂量即明显下降。细胞培养液中LDH活性、TNF-α释放量、IL-6释放量基本上随着染尘剂量的升高和作用时间的延长而逐渐升高。随着作用的时间延长，细胞因子TNF-α和IL-6开始出现明显变化的剂量有逐渐降低的趋势，反之亦然；LDH活性变化的剂量则未见这个趋势，明显变化的剂量始终在10μg/ml。 SOD活力的变化与上述指标不同。在6、12h时间点，随着染尘剂量增加，SOD活力呈升高趋势，在24h时间点，，SOD活力呈先增高后降低的改变，在48h时间点，SOD活力呈降低趋势。随着作用时间的延长，SOD活力呈现先升高后降低的趋势。结论: Nano-SiO2能够对HUVECs产生一定的毒性效应，且对细胞的毒性效应存在明显的剂量和时间依赖性。
[Abstract]:Objective: occupational safety and health problems of nanomaterials are complex. Nanomaterials have a large surface area, although they are small in size, resulting in great changes in physical, chemical and biological properties compared with those of larger particles of the same composition. These physical and chemical properties are likely to affect the ability of nanomaterials to reach the lung gas exchange region, transport from the lung to the whole body, penetrate the skin barrier, cross the cell membrane, and even react at the molecular level, etc. Some preliminary research results show that the safety evaluation results of micron materials may not be applicable to nanoscale materials, so the biosafety of nanoscale materials has been paid more and more attention. Therefore, in this study, typical nano-particles (SiO2) were selected as dust particles, and the dose and time effects of cardiovascular toxicity of nanoparticles were studied in vitro. To provide further experimental basis for cardiovascular safety evaluation of nano-scale chemicals. Methods: human umbilical vein endothelial cells (HUVECs) were selected as dust-contaminated particles and human umbilical vein endothelial cells (HUVECs) were co-cultured with human umbilical vein endothelial cells (HUVECs) at different doses for 6 ~ 12 ~ 24 ~ (24) h for different time. The survival rate of cells was measured by CCK-8 method, and the lethal dose of HUVECs was calculated by 50 渭 g / ml of human umbilical vein endothelial cells (HUVECs). The dose of dust in the follow-up experiment was determined to be 0 ~ 5 ~ 10 ~ 10 ~ 20 渭 g 路ml ~ (-1) 路L ~ (-1). The above dosages of nano-SiO2 particles were co-cultured with HUVECs at a dose of 10 ~ 10 渭 g / ml and 60 渭 g / ml, respectively, and then cultured for 48 h with 6o _ (12) O _ (24) O _ (24) 渭 g / ml. The activity of lactate dehydrogenase (LDH), the activity of total superoxide dismutase (SOD), the release of tumor necrosis factor- 伪 (TNF- 伪) and the release of interleukin-6 (IL-6) in the supernatant of cell culture medium were measured by the corresponding kit. Results: the cell survival rate decreased gradually with the increase of nano-SiO2 dust exposure dose and the prolongation of the time of action. If the time was short, the cell survival rate began to decrease at a higher dose, and if the time was long, the cell survival rate decreased significantly at a low dose. The release of TNF- 伪 and IL-6 in the cell culture medium increased gradually with the increase of dust exposure dose and the prolongation of the time of action. The dosages of TNF- 伪 and IL-6 began to change gradually with the prolongation of the time of action, but the dose of the change of activity of TNF- 伪 and IL-6 was not seen at the same time, and the dose of obvious change was always at 10 渭 g / ml. The change of SOD activity was different from the above indexes. At 6h / 12h, the SOD activity increased with the increase of dust exposure dose, increased first and then decreased at 24h, and decreased at 48h. The activity of SOD increased first and then decreased with the prolongation of action time. Conclusion: Nano-SiO2 can produce a certain toxic effect on HUVECs, and its cytotoxic effect is dose and time dependent.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114

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