结核杆菌对感染小鼠肺泡巨噬细胞铁代谢的调控作用及其机制研究

发布时间：2018-06-24 06:19

本文选题：铁蛋白 + 转铁蛋白受体　；参考：《石河子大学》2014年硕士论文

【摘要】：目的：通过构建结核分枝杆菌感染小鼠肺泡巨噬细胞模型，探讨研究不同毒力的结核分枝杆菌对小鼠肺泡巨噬细胞铁代谢调控作用机制。方法：分别将结核分枝杆菌国际标准强毒株H37Rv菌株（H37Rv）、卡介苗菌株（BCG）以及无菌生理盐水（空白对照）通过小鼠尾静脉，，注入小鼠体内建立小鼠的感染模型，并在感染后的1d、3d、5d、7d、9d、11d、13d、15d提取小鼠肺泡巨噬细胞总蛋白，利用ELISA方法于模型建立成功后1d、3d、5d、7d、9d、11d、13d、15d检测各组小鼠肺泡巨噬细胞内铁蛋白、转铁蛋白受体、膜铁转运蛋白、乳铁蛋白的表达，并分析其时相性变化，应用Western blot技术比较分析各组小鼠肺泡巨噬细胞内铁蛋白、转铁蛋白受体于上述各时间点蛋白表达水平的时相性变化。结果： 1.感染组小鼠肺泡巨噬细胞内Fn表达量随时间延长逐渐降低，在感染后7d、9d、11d最低，差异明显，具有统计学意义（P0.05）。 2.感染导致小鼠肺泡巨噬细胞内的TfR表达量逐渐增高，于感染后7d、9d、11d最高，并且H37Rv组高于BCG组表达量，差异具有统计学意义（P0.05）。 3.感染后的小鼠肺泡巨噬细胞内FPN的表达量逐渐降低，于感染后5d、7d、9d最明显，差异具有统计学意义（P0.05）。 4.感染导致小鼠肺泡巨噬细胞分泌较高水平的Lf，与对照组比较在感染后7d、9d、11d升高最明显，差异均具有统计学意义（P0.05）。结论： 1.小鼠肺泡巨噬细胞内Fn含量下降，提示感染导致细胞内铁离子含量的下降。 2.小鼠肺泡巨噬细胞巨噬细胞内TfR及Lf含量的升高，提示结核杆菌的感染导致巨噬细胞内摄铁通路的开放。 3.小鼠肺泡巨噬细胞巨噬细胞内FPN含量下降，提示结核杆菌的感染导致巨噬细胞内铁释放通路的关闭。
[Abstract]:Objective: to study the regulatory mechanism of different virulent mycobacterium tuberculosis on iron metabolism of alveolar macrophages in mice by constructing a mouse alveolar macrophage model infected with Mycobacterium tuberculosis. Methods: Mycobacterium tuberculosis strain H37Rv (International Standard strain H37Rv), Bacillus Calmette-Guerin strain (BCG) and sterile saline (blank control) were injected into the tail vein of mice to establish the model of infection. The total protein of mouse alveolar macrophages was extracted from mice alveolar macrophages at 1 day, 3 days, 5 days, 7 days, 9 days and 9 days after infection. The expression of ferritin, transferrin receptor, membrane ferritin transporter and lactoferrin in alveolar macrophages of each group were detected by Elisa, and the expression of ferritin, transferrin receptor, membrane ferritin transporter and lactoferrin in alveolar macrophages of each group were detected by Elisa, and the expression of ferritin, transferrin receptor, membrane ferritin receptor and lactoferrin in alveolar macrophages of each group were detected by Elisa. The temporal changes of the expression of ferritin and transferrin receptor in alveolar macrophages were analyzed by Western blot. Results: 1. The expression of FN in alveolar macrophages of infected mice decreased gradually with the prolongation of time, and was the lowest at 7 days and 9 days after infection, and the difference was significant (P0.05). The expression of TfR in alveolar macrophages increased gradually after infection, and reached the highest level at day 11d after infection, and the expression of TfR in H37Rv group was higher than that in BCG group (P0.05). 3. The expression of TfR in alveolar macrophages was significantly higher in H37Rv group than that in BCG group (P0.05). The expression of FPN in alveolar macrophages of infected mice decreased gradually, and the expression of FPN in alveolar macrophages was the most obvious on the 5th day and 7th day after infection, and the difference was statistically significant (P0.05). Compared with the control group, the level of Lfsecreted by mouse alveolar macrophages was significantly higher than that of the control group on the 7th day and 9th day after infection, and the difference was statistically significant (P0.05). Conclusion: 1. The content of FN in alveolar macrophages in mice was decreased, indicating that the iron ion content in alveolar macrophages decreased. 2. 2. The increase of TfR and LF contents in alveolar macrophages of mice suggests that the infection of Mycobacterium tuberculosis leads to the opening of TieTong pathway in macrophages. The decrease of FPN content in alveolar macrophages suggested that the iron release pathway in macrophages was shut down due to the infection of Mycobacterium tuberculosis.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R52

【参考文献】