铀矿尘对巨噬细胞毒性作用及TGF-β1表达的影响

发布时间：2018-03-05 20:33

本文选题：铀矿尘　切入点：巨噬细胞　出处：《南华大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：探讨铀矿尘作用小鼠巨噬细胞不同时间后的细胞毒性作用及铀矿尘诱导巨噬细胞表达TGF-β1的影响。方法：采用小鼠巨噬细胞系RAW264.7细胞株，进行体外细胞培养实验，实验分别设铀矿尘实验组和无粉尘培养基对照组。实验组用120μg/ml的铀矿尘混悬液与RAW264.7细胞分别作用2、4、8、16、24、32h后，通过MTT实验、HE染色、AO/EB染色等方法观察铀矿尘对RAW264.7增殖活性、细胞形态及细胞凋亡的影响；利用比色法测定细胞培养上清液中·OH、H2O2及MDA水平的变化；应用ELISA法测定铀矿尘对RAW264.7细胞培养上清液中TGF-β1水平的影响。结果：铀矿尘混悬液作用RAW264.7细胞2、4、8、16、24、32h后，MTT实验结果显示，RAW264.7细胞增殖活性明显降低（与对照组比较，P＜0.05），并且随着染尘时间的延长，RAW264.7细胞的存活率逐渐降低；HE染色可见，随着染尘时间的延长，铀矿尘组梭型或不规则型细胞数目上升，多核细胞数明显增加，细胞胞质缺失，胞核外露、染色变浅，细胞形态变化较大；AO/EB染色发现，铀矿尘组红色或橙色的凋亡/坏死细胞明显增多，并且随着染尘时间的延长，坏死和凋亡细胞数目逐渐增加；比色法测定细胞培养上清液中·OH、H2O2及MDA含量发现，铀矿尘组细胞培养上清液中·OH、H2O2、MDA水平明显升高（与对照组比较，P＜0.05），并且随着染尘时间的延长，，其水平持续升高；ELISA实验结果显示，铀矿尘能够引起RAW264.7细胞培养上清液中TGF-β1水平增高（与对照组比较，P＜0.05），而且随着染尘时间的延长，TGF-β1水平逐渐增高，在24h时达到峰值。结论： 1、铀矿尘能够引起巨噬细胞凋亡或崩解，抑制细胞增殖活性。 2、铀矿尘可以引起细胞培养上清液中自由基水平升高，并诱发脂质过氧化反应。 3、铀矿尘能够诱导巨噬细胞分泌TGF-β1，引起细胞培养上清液中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.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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