本文選題：鈾礦塵　切入點(diǎn)：巨噬細(xì)胞　出處：《南華大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探討鈾礦塵作用小鼠巨噬細(xì)胞不同時(shí)間后的細(xì)胞毒性作用及鈾礦塵誘導(dǎo)巨噬細(xì)胞表達(dá)TGF-β1的影響。 方法：采用小鼠巨噬細(xì)胞系RAW264.7細(xì)胞株，進(jìn)行體外細(xì)胞培養(yǎng)實(shí)驗(yàn)，實(shí)驗(yàn)分別設(shè)鈾礦塵實(shí)驗(yàn)組和無粉塵培養(yǎng)基對(duì)照組。實(shí)驗(yàn)組用120μg/ml的鈾礦塵混懸液與RAW264.7細(xì)胞分別作用2、4、8、16、24、32h后，通過MTT實(shí)驗(yàn)、HE染色、AO/EB染色等方法觀察鈾礦塵對(duì)RAW264.7增殖活性、細(xì)胞形態(tài)及細(xì)胞凋亡的影響；利用比色法測定細(xì)胞培養(yǎng)上清液中·OH、H2O2及MDA水平的變化；應(yīng)用ELISA法測定鈾礦塵對(duì)RAW264.7細(xì)胞培養(yǎng)上清液中TGF-β1水平的影響。 結(jié)果：鈾礦塵混懸液作用RAW264.7細(xì)胞2、4、8、16、24、32h后，MTT實(shí)驗(yàn)結(jié)果顯示，RAW264.7細(xì)胞增殖活性明顯降低（與對(duì)照組比較，P＜0.05），并且隨著染塵時(shí)間的延長，RAW264.7細(xì)胞的存活率逐漸降低；HE染色可見，隨著染塵時(shí)間的延長，鈾礦塵組梭型或不規(guī)則型細(xì)胞數(shù)目上升，多核細(xì)胞數(shù)明顯增加，細(xì)胞胞質(zhì)缺失，胞核外露、染色變淺，細(xì)胞形態(tài)變化較大；AO/EB染色發(fā)現(xiàn)，鈾礦塵組紅色或橙色的凋亡/壞死細(xì)胞明顯增多，并且隨著染塵時(shí)間的延長，壞死和凋亡細(xì)胞數(shù)目逐漸增加；比色法測定細(xì)胞培養(yǎng)上清液中·OH、H2O2及MDA含量發(fā)現(xiàn)，鈾礦塵組細(xì)胞培養(yǎng)上清液中·OH、H2O2、MDA水平明顯升高（與對(duì)照組比較，P＜0.05），并且隨著染塵時(shí)間的延長，，其水平持續(xù)升高；ELISA實(shí)驗(yàn)結(jié)果顯示，鈾礦塵能夠引起RAW264.7細(xì)胞培養(yǎng)上清液中TGF-β1水平增高（與對(duì)照組比較，P＜0.05），而且隨著染塵時(shí)間的延長，TGF-β1水平逐漸增高，在24h時(shí)達(dá)到峰值。 結(jié)論： 1、鈾礦塵能夠引起巨噬細(xì)胞凋亡或崩解，抑制細(xì)胞增殖活性。 2、鈾礦塵可以引起細(xì)胞培養(yǎng)上清液中自由基水平升高，并誘發(fā)脂質(zhì)過氧化反應(yīng)。 3、鈾礦塵能夠誘導(dǎo)巨噬細(xì)胞分泌TGF-β1，引起細(xì)胞培養(yǎng)上清液中TGF-β1水平升高。
[Abstract]:Aim: to investigate the cytotoxicity of mouse macrophages induced by uranium dust and the effect of uranium dust on the expression of TGF- 尾 1 in macrophages. Methods: a mouse macrophage cell line RAW264.7 cell line was used for cell culture in vitro. The experimental group was divided into two groups: uranium mine dust test group and dust free medium control group. The experimental group was treated with 120 渭 g / ml uranium ore dust suspension and RAW264.7 cells for 2432 hours, respectively. The effects of uranium dust on the proliferation activity, cell morphology and apoptosis of RAW264.7 were observed by means of MTT and HE staining, and the changes of H _ 2O _ 2 and MDA levels in the supernatant of cell culture were determined by colorimetry. The effect of uranium dust on the level of TGF- 尾 1 in supernatant of RAW264.7 cell culture was determined by ELISA assay. Results: the results showed that the proliferative activity of RAW264.7 cells decreased significantly (P < 0.05) compared with the control group, and the survival rate of RAW264.7 cells decreased gradually with the prolongation of dust exposure time, and the survival rate of RAW264.7 cells decreased gradually with the prolongation of dust exposure time, the results showed that the proliferation activity of RAW264.7 cells was significantly lower than that of the control group (P < 0.05), and the survival rate of RAW264.7 cells decreased gradually with the prolongation of dust exposure time. With the prolongation of the time of dust exposure, the number of fusiform or irregular type cells in uranium dust group increased, the number of multinucleated cells increased obviously, the cytoplasm of the cells was absent, the cytoplasm was exposed, the staining became shallower, and the morphological changes of the cells were found by AOP / EB staining. The number of apoptotic / necrotic cells increased with the prolongation of dust exposure time, and the contents of 路OHH _ 2O _ 2 and MDA in the supernatant of cell culture were determined by colorimetric method. The level of MDA in the supernatant of cell culture of uranium mine dust group was significantly higher than that in the control group (P < 0.05), and with the prolongation of dust exposure time, the level of MDA in the supernatant of cell culture was increased continuously. Uranium dust could increase the level of TGF- 尾 1 in the supernatant of RAW264.7 cell culture (compared with the control group, P < 0.05), and the level of TGF- 尾 1 increased gradually with the prolongation of dust exposure time, and reached its peak at 24 h. Conclusion:. 1. Uranium dust can induce apoptosis or disintegration of macrophages and inhibit cell proliferation. 2. Uranium dust can increase the level of free radical and induce lipid peroxidation in the supernatant of cell culture. 3. Uranium dust could induce macrophages to secrete TGF- 尾 1 and increase the level of TGF- 尾 1 in the supernatant of cell culture.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R363

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