發(fā)布時間：2018-04-20 19:28

  本文選題：高硅質(zhì) + 細顆粒物��； 參考：《西南醫(yī)科大學》2017年碩士論文

【摘要】：目的:近年來霧霾天氣愈漸突出,石英和納米二氧化硅是霧霾天氣中大氣顆粒物的兩種主要富硅細顆粒礦物粉塵。本文以這兩種粉塵為實驗對象,研究及分析兩者對人肺腺癌A549細胞的毒性作用。方法:(1)50μg/mL、100μg/mL、200μg/mL、400μg/mL和800μg/mL的石英和納米二氧化硅兩種粉塵懸液,分別染毒A549細胞24 h,采用MTT試驗分析這兩種粉塵對A549細胞存活率的影響。(2)50μg/mL、200μg/mL和800μg/mL的石英和納米二氧化硅兩種粉塵懸液,分別染毒A549細胞24 h,檢測培養(yǎng)上清液中乳酸脫氫酶(lactate dehydrogenase,LDH)活力,分析不同濃度的這兩種粉塵對A549細胞膜通透性的影響差異;200μg/mL的石英和納米二氧化硅兩種礦物粉塵懸液,分別染毒A549細胞3 h、6 h和18 h,檢測培養(yǎng)上清液中LDH活力,分析這兩種粉塵對A549細胞膜通透性的影響隨染毒時間的變化趨勢。(3)50μg/mL、200μg/mL和800μg/mL的石英和納米二氧化硅兩種礦物粉塵懸液,分別于染毒A549細胞2 h、4 h、8 h、16 h、24 h和48 h,檢測細胞內(nèi)超氧化物歧化酶(superoxide dismutase,SOD)活力和谷胱甘肽酶(glutathione,GSH)含量。(4)流式細胞儀PI染色分析這兩種富硅細顆粒礦物粉塵對A549細胞周期及凋亡的影響。結(jié)果:(1)在實驗設(shè)定的50μg/mL-800μg/mL濃度范圍內(nèi),與陰性對照組相比較,隨著兩種礦物粉塵懸液濃度的增高,A549細胞存活率降低,呈現(xiàn)較好的劑量-效應關(guān)系。納米二氧化硅引起細胞的存活率降低程度大于石英。(2)兩種礦物粉塵染毒A549細胞24 h,細胞上清液中LDH的活力均增加,并隨礦物粉塵懸液濃度的增加,LDH的活力呈現(xiàn)增加的趨勢;隨著兩種礦物粉塵懸液染毒時間延長,LDH的活力呈現(xiàn)增加的趨勢;納米二氧化硅引起A549細胞的LDH釋放量大于石英。(3)在50μg/mL-800μg/mL濃度范圍內(nèi),隨著礦物粉塵濃度的升高,A549細胞的SOD活力增高,GSH含量降低。(4)石英和納米二氧化硅兩種富硅細顆粒礦物粉塵均能夠誘導A549細胞G2期阻滯和引起部分細胞凋亡。結(jié)論:石英和納米二氧化硅兩種富硅細顆粒礦物粉塵均能夠破壞A549細胞膜完整性,引起細胞抗氧化體系指標改變,誘導A549細胞G2期阻滯和部分細胞凋亡,納米二氧化硅粉塵對A549細胞的毒性作用強于石英粉塵。
[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.
【學位授予單位】：西南醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R994.6

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