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首页> 外文期刊>American Journal of Physiology >Interferon-gamma decreases barrier function in T84 cells by reducing ZO-1 levels and disrupting apical actin.
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Interferon-gamma decreases barrier function in T84 cells by reducing ZO-1 levels and disrupting apical actin.

机译：γ干扰素可通过降低ZO-1水平并破坏顶端肌动蛋白来降低T84细胞的屏障功能。

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The effects of interferon-gamma (IFN-gamma) on tight junctions in T84 human intestinal epithelial cells were investigated. Treatment of T84 cells with IFN-gamma caused a dose- and time-dependent increase in monolayer permeability as assessed by transepithelial electrical resistance measurements. Examination of specific proteins associated with tight junctions by immunoblotting and confocal microscopy revealed changes in the expression levels and localization of some of these proteins after exposure of the cells to IFN-gamma. Specifically, IFN-gamma treatment resulted in an almost total loss of zonula occludens (ZO)-1, whereas the levels of ZO-2 and occludin showed relatively modest decreases compared with untreated cells. Loss of ZO-1 was associated with the altered localization of ZO-2 and occludin. In IFN-gamma-treated cells, ZO-2 and occludin were diffusely distributed, whereas, in control cells, they, along with ZO-1, were predominantly localized to the tight junctions. Analysis of ZO-1 protein and RNA by pulse chase and RT-PCR, respectively, showed an increase in protein turnover, a decrease in protein synthesis, and a reduction in RNA levels following IFN-gamma treatment. In contrast to ZO-1, ZO-2 and occludin did not show any major changes in these parameters. In addition, the organization of actin in the apical and tight junction regions, but not in the mid- or basal regions, of the cells was also perturbed by IFN-gamma treatment of cells. Time-course analysis of IFN-gamma-induced alterations in ZO-1 expression and apical actin perturbation indicated that these two effects were intimately linked and could not be dissociated. These results suggest that IFN-gamma affects barrier function in T84 cells by decreasing the levels of ZO-1 and perturbing apical actin organization, which leads to a disorganization of the tight junction and an increase in paracellular permeability.
机译：研究了干扰素-γ（IFN-γ）对人T84肠上皮细胞紧密连接的影响。通过经上皮电阻测量评估，用IFN-γ处理T84细胞导致单层渗透性的剂量和时间依赖性增加。通过免疫印迹和共聚焦显微镜检查与紧密连接相关的特定蛋白质，发现细胞暴露于IFN-γ后表达水平发生变化，某些蛋白质的定位也发生了变化。具体而言，IFN-γ处理导致小带闭合（ZO）-1几乎完全丧失，而与未处理的细胞相比，ZO-2和闭合蛋白的水平显示相对适度的下降。 ZO-1的丢失与ZO-2和occludin的定位改变有关。在IFN-γ处理的细胞中，ZO-2和occludin分散分布，而在对照细胞中，它们与ZO-1一起主要位于紧密连接处。分别通过脉冲追踪和RT-PCR对ZO-1蛋白和RNA进行分析，结果表明，经过IFN-γ处理后，蛋白更新量增加，蛋白合成减少，RNA水平降低。与ZO-1相反，ZO-2和occludin在这些参数上没有显示任何重大变化。另外，肌动蛋白在细胞的顶端和紧密连接区域中而不是在中部或基底区域中的组织也受到IFN-γ处理细胞的干扰。 IFN-γ诱导的ZO-1表达变化和心尖肌动蛋白扰动的时程分析表明，这两种作用密切相关，无法分离。这些结果表明，IFN-γ通过降低ZO-1的水平和扰动顶端肌动蛋白组织来影响T84细胞的屏障功能，从而导致紧密连接的混乱和旁细胞通透性的增加。

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