两种富硅细颗粒矿物粉尘对人肺腺癌A549细胞的毒性作用研究

发布时间：2018-04-20 19:28

本文选题：高硅质 + 细颗粒物　；参考：《西南医科大学》2017年硕士论文

【摘要】：目的:近年来雾霾天气愈渐突出,石英和纳米二氧化硅是雾霾天气中大气颗粒物的两种主要富硅细颗粒矿物粉尘。本文以这两种粉尘为实验对象,研究及分析两者对人肺腺癌A549细胞的毒性作用。方法:(1)50μg/mL、100μg/mL、200μg/mL、400μg/mL和800μg/mL的石英和纳米二氧化硅两种粉尘悬液,分别染毒A549细胞24 h,采用MTT试验分析这两种粉尘对A549细胞存活率的影响。(2)50μg/mL、200μg/mL和800μg/mL的石英和纳米二氧化硅两种粉尘悬液,分别染毒A549细胞24 h,检测培养上清液中乳酸脱氢酶(lactate dehydrogenase,LDH)活力,分析不同浓度的这两种粉尘对A549细胞膜通透性的影响差异;200μg/mL的石英和纳米二氧化硅两种矿物粉尘悬液,分别染毒A549细胞3 h、6 h和18 h,检测培养上清液中LDH活力,分析这两种粉尘对A549细胞膜通透性的影响随染毒时间的变化趋势。(3)50μg/mL、200μg/mL和800μg/mL的石英和纳米二氧化硅两种矿物粉尘悬液,分别于染毒A549细胞2 h、4 h、8 h、16 h、24 h和48 h,检测细胞内超氧化物歧化酶(superoxide dismutase,SOD)活力和谷胱甘肽酶(glutathione,GSH)含量。(4)流式细胞仪PI染色分析这两种富硅细颗粒矿物粉尘对A549细胞周期及凋亡的影响。结果:(1)在实验设定的50μg/mL-800μg/mL浓度范围内,与阴性对照组相比较,随着两种矿物粉尘悬液浓度的增高,A549细胞存活率降低,呈现较好的剂量-效应关系。纳米二氧化硅引起细胞的存活率降低程度大于石英。(2)两种矿物粉尘染毒A549细胞24 h,细胞上清液中LDH的活力均增加,并随矿物粉尘悬液浓度的增加,LDH的活力呈现增加的趋势;随着两种矿物粉尘悬液染毒时间延长,LDH的活力呈现增加的趋势;纳米二氧化硅引起A549细胞的LDH释放量大于石英。(3)在50μg/mL-800μg/mL浓度范围内,随着矿物粉尘浓度的升高,A549细胞的SOD活力增高,GSH含量降低。(4)石英和纳米二氧化硅两种富硅细颗粒矿物粉尘均能够诱导A549细胞G2期阻滞和引起部分细胞凋亡。结论:石英和纳米二氧化硅两种富硅细颗粒矿物粉尘均能够破坏A549细胞膜完整性,引起细胞抗氧化体系指标改变,诱导A549细胞G2期阻滞和部分细胞凋亡,纳米二氧化硅粉尘对A549细胞的毒性作用强于石英粉尘。
[Abstract]:Objective: in recent years, the weather of haze has become more and more prominent. Quartz and nano-silica are the two main kinds of silicon-rich fine particulate mineral dust in haze weather. The toxic effects of these two dusts on human lung adenocarcinoma cell line A549 were studied and analyzed. Methods two kinds of dust suspensions of 200 渭 g 路mL ~ (-1) 100 渭 g 路mL ~ (-1) ~ 100 渭 g 路mL ~ (-1) ~ 100 渭 g 路mL ~ (-1) ~ 100 渭 g 路mL ~ (-1) and 400 渭 g/mL and 800 渭 g/mL silica were exposed to A549 cells for 24 h, respectively. The effects of these two kinds of dust on the survival rate of A549 cells were analyzed by MTT test. The suspensions of silica (200 渭 g/mL) and silica (800 渭 g/mL) of 200 渭 g 路mL ~ (-1) and 800 渭 g/mL were used to analyze the survival rate of A549 cells. The activity of lactate dehydrogenase (LDH) in the supernatant of A549 cells was detected for 24 h, and the effects of different concentrations of dust on membrane permeability of A549 cells were analyzed. The mineral dust suspensions of quartz and nano-silica at 200 渭 g/mL were analyzed. A549 cells were exposed to A549 cells for 3 h and 18 h, respectively. The activity of LDH in the supernatant was detected, and the effect of these two kinds of dust on the membrane permeability of A549 cells was analyzed. The effect of these two kinds of dust on the membrane permeability of A549 cells was analyzed with the time of exposure. The mineral dust suspensions of quartz and nanometer silica of 200 渭 g/mL and 800 渭 g/mL were analyzed. The activity of superoxide dismutase (SOD) and glutathione glutathione glutathione (GSH) content in A549 cells were detected at 2 h, 4 h, 8 h, 16 h and 48 h, respectively. Flow cytometry (FCM) was used to analyze the effects of these silicon-rich fine particulate mineral dust on A549 cells. Phase and apoptosis. Results in the concentration range of 50 渭 g/mL-800 渭 g/mL, compared with the negative control group, the survival rate of A549 cells decreased with the increase of the concentration of two mineral dust suspensions, and showed a good dose-effect relationship. The activity of LDH in the supernatant of A549 cells was increased with the increase of the concentration of mineral dust, and the activity of LDH increased with the increase of the concentration of mineral dust. The LDH release of A549 cells induced by nano-silica was larger than that of quartz. The concentration of LDH in A549 cells was higher than that of silica in the concentration range of 50 渭 g/mL-800 渭 g/mL. With the increase of mineral dust concentration, the SOD activity of A549 cells increased and the content of SOD decreased. 4) Quartz and nano-silica two kinds of fine silicon-rich mineral dust could block the G2 phase of A549 cells and induce some cell apoptosis. Conclusion: both silica and silica particles can destroy the integrity of A549 cell membrane, change the index of antioxidant system, and induce G2 arrest and apoptosis of A549 cells. The toxicity of nano silica dust to A549 cells was stronger than that of quartz dust.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R994.6

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