本文選題：三氧化二砷 + 納米��； 參考：《吉林大學》2014年碩士論文

【摘要】：本實驗采用溶膠-凝膠法制備As2O3納米粒，通過MTT法比較As2O3納米粒和As2O3對NB4細胞的增殖抑制作用；通過Hoechst和PI染色、流式細胞儀（Flow cytometry,FCM）觀察檢測As2O3納米粒和As2O3對NB4細胞的凋亡作用；通過羅丹明123染色檢測兩種劑型對NB4細胞的線粒體膜電位的影響；通過Western blot觀察兩種劑型作用NB4細胞后細胞內(nèi)p-PTEN、p-AKT、Bax、caspase-3、caspase-9和AIF的表達變化，初步探討As2O3納米粒對NB4細胞的增殖抑制和誘導凋亡的作用機制。 細胞增殖抑制實驗結果顯示As2O3納米粒和As2O3對NB4細胞均可產(chǎn)生抑制作用，且呈時間濃度依賴性；藥物濃度和作用時間相同時，As2O3納米粒對NB4細胞的生長抑制作用明顯強于As2O3。細胞凋亡實驗結果顯示，As2O3納米粒和As2O3作用NB4細胞后主要表現(xiàn)為凋亡，相同作用時間下，隨藥物濃度增加，凋亡率增加，且納米粒組凋亡率明顯多于傳統(tǒng)劑型As2O3組。羅丹明123染色實驗結果顯示兩種劑型均可降低NB4細胞的線粒體膜電位，且呈濃度依賴性；相同藥物濃度和作用時間下，，As2O3納米粒對NB4細胞的作用明顯強于As2O3。Western blot結果顯示，與同濃度As2O3比較，1.5μmol/L、3μmol/L As2O3納米粒能顯著下調(diào)p-AKT和PTEN的表達；相同濃度時，As2O3納米粒較As2O3明顯增加p-PTEN和Bax的表達，且呈濃度依賴性；1.5μmol/L和3μmol/LAs2O3caspase-3、caspase-9的表達量呈濃度依賴性增加，1.5μmol/L As2O3納米粒較同濃度三氧化二砷表達量增加，而3μmol/LAs2O3納米粒組caspase-3、caspase-9的表達量較同濃度三氧化二砷降低。隨后繼續(xù)研究了AIF的表達變化，結果顯示3μmol/L As2O3納米粒組AIF的表達量增加。因此，根據(jù)實驗結果推測三氧化二砷和低濃度三氧化二砷納米粒主要通過依賴caspase的線粒體凋亡途徑誘導NB4細胞凋亡，而高濃度三氧化二砷納米粒主要通過非依賴caspase的線粒體凋亡途徑誘導NB4細胞凋亡。 根據(jù)上述實驗結果得出以下結論： ⑴As2O3納米粒和As2O3對NB4細胞的生長有抑制作用，呈時間和濃度依賴性，且As2O3納米粒對NB4細胞的作用明顯強于As2O3； ⑵As2O3納米粒和As2O3誘導NB4細胞凋亡呈濃度依賴性；且As2O3納米粒的誘導凋亡作用更為顯著； ⑶As2O3納米粒對NB4細胞誘導凋亡作用強于As2O3，可能與上調(diào)p-PTEN的表達、抑制AKT的活化和啟動線粒體凋亡途徑有關。
[Abstract]:In this experiment, As2O3 nanoparticles were prepared by sol-gel method, and the inhibitory effects of As2O3 nanoparticles and As2O3 on the proliferation of NB4 cells were compared by MTT method, and the apoptotic effects of As2O3 nanoparticles and As2O3 on NB4 cells were observed by flow cytometry (FCM) and Hoechst and Pi staining. The effects of two dosage forms on mitochondrial membrane potential of NB4 cells were detected by rhodamine 123 staining, and the changes of expression of p-PTEN, p-AKTX, caspase-3, caspase-9 and AIF in NB4 cells were observed by Western blot. To explore the mechanism of As2O3 nanoparticles inhibiting proliferation and inducing apoptosis of NB4 cells. The results of cell proliferation inhibition test showed that both As2O3 nanoparticles and As2O3 could inhibit the growth of NB4 cells in a time and concentration dependent manner, and the inhibitory effect of As2O3 nanoparticles on the growth of NB4 cells was stronger than that of as 2O 3 at the same drug concentration and time. The results of apoptosis experiment showed that the apoptosis rate of NB4 cells was increased with the increase of drug concentration at the same time, and the apoptotic rate of As2O3 group was higher than that of As2O3 group. The results of Rhodamine 123 staining showed that the mitochondrial membrane potential of NB4 cells was decreased in a concentration-dependent manner, and the effect of as _ 2O _ 3 nanoparticles on NB4 cells was significantly stronger than that of As2O3.Western blot at the same concentration and time. Compared with the same concentration of As2O3, 1.5 渭 mol 路L ~ (-3) 渭 mol/L As2O3 nanoparticles could significantly down-regulate the expression of p-AKT and PTEN, and the same concentration of as _ 2O _ 3 nanoparticles could significantly increase the expression of p-PTEN and Bax in the same concentration of as _ 2O _ 3 nanoparticles compared with As2O3. In a dose-dependent manner, the expression of caspase-3 caspase-9 was increased in a dose-dependent manner by 1.5 渭 mol/L and 3 渭 mol / L as _ 2O _ 3 caspase-3 caspase-9 in a dose-dependent manner, and the expression of caspase-3 caspase-9 in 3 渭 mol/LAs2O3 nanoparticles was lower than that in the same concentration of arsenic trioxide. The expression of AIF was further studied. The results showed that the expression of AIF in 3 渭 mol/L As2O3 nanoparticles was increased. Therefore, according to the experimental results, it is inferred that arsenic trioxide nanoparticles and low concentration arsenic trioxide nanoparticles induce apoptosis of NB4 cells mainly through mitochondrial apoptosis pathway dependent on caspase. High concentration of arsenic trioxide nanoparticles mainly induce apoptosis of NB4 cells through mitochondrial apoptosis independent of caspase. Based on the above experimental results, the following conclusions are drawn: 1As2O3 nanoparticles and As2O3 inhibited the growth of NB4 cells in a time-and concentration-dependent manner, and the effect of As2O3 nanoparticles on NB4 cells was significantly stronger than that on as _ 2O _ 3 cells. The apoptosis of NB4 cells induced by 2As2O3 nanoparticles and As2O3 was concentration-dependent, and the apoptosis induced by As2O3 nanoparticles was more significant. The effect of 3As2O3 nanoparticles on apoptosis of NB4 cells is stronger than that of As2O3, which may be related to up-regulation of p-PTEN expression, inhibition of AKT activation and activation of mitochondrial apoptosis pathway.
【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R943;R96

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