Diesel exhaust particles modulate the tight junction protein occludin in lung cells in vitro
Diesel exhaust particles modulate the tight junction protein occludin in lung cells in vitro
复制标题
DOI：
10.1186/1743-8977-6-26
复制
发表时间：
2009-10-08
影响因子：
10
通讯作者：
Rothen-Rutishauser, Barbara M.
中科院分区：
文献类型：
--
作者：
Lehmann, Andrea D.;Blank, Fabian;Rothen-Rutishauser, Barbara M.
背景：利用由人上皮细胞(16HBE140-)、单核细胞来源的巨噬细胞和树突状细胞组成的体外三重细胞共培养模型，最近证明巨噬细胞和树突状细胞在上皮细胞之间建立跨上皮网络，在不破坏上皮紧密性的情况下捕获抗原。巨噬细胞和树突状细胞表达不同的紧密连接蛋白，并与上皮细胞形成紧密连接样结构。采用免疫荧光技术结合激光扫描显微镜和实时定量聚合酶链式反应技术，观察柴油车尾气颗粒物(0.5、5、50、125mU g/ml)作用24 h后，3种细胞紧密连接蛋白表达的变化。结果：只有最大剂量(125mU g/ml)的柴油尾气颗粒物(DEP)能抑制防御系统细胞中的occludin基因的表达，但对上皮细胞中的occludin基因表达无明显影响。在接触高浓度DEP后，跨上皮细胞的电阻在单细胞培养中降低，但在三细胞共培养中并不降低。结论：高浓度的DEP(125mU g/ml)可调节肺防御系统细胞紧密连接闭锁蛋白的表达，提示肺细胞暴露于颗粒性抗原后，肺上皮细胞在维持上皮细胞完整性中起重要作用。
Background: Using an in vitro triple cell co-culture model consisting of human epithelial cells (16HBE14o-), monocyte-derived macrophages and dendritic cells, it was recently demonstrated that macrophages and dendritic cells create a transepithelial network between the epithelial cells to capture antigens without disrupting the epithelial tightness. The expression of the different tight junction proteins in macrophages and dendritic cells, and the formation of tight junction-like structures with epithelial cells has been demonstrated. Immunofluorescent methods combined with laser scanning microscopy and quantitative real-time polymerase chain reaction were used to investigate if exposure to diesel exhaust particles (DEP) (0.5, 5, 50, 125 mu g/ml), for 24 h, can modulate the expression of the tight junction mRNA/protein of occludin, in all three cell types.Results: Only the highest dose of DEP (125 mu g/ml) seemed to reduce the occludin mRNA in the cells of the defence system however not in epithelial cells, although the occludin arrangement in the latter cell type was disrupted. The transepithelial electrical resistance was reduced in epithelial cell mono-cultures but not in the triple cell co-cultures, following exposure to high DEP concentration. Cytotoxicity was not found, in either epithelial mono-cultures nor in triple cell co-cultures, after exposure to the different DEP concentrations.Conclusion: We concluded that high concentrations of DEP (125 mu g/ml) can modulate the tight junction occludin mRNA in the cells of the defence system and that those cells play an important role maintaining the epithelial integrity following exposure to particulate antigens in lung cells.