Zhang Yong-Guo, Wu Shaoping, Xia Yinglin, Sun Jun
Department of Biochemistry, Rush University, 1735 W. Harrison St., Chicago, Illinois.
Department of Biostatistics and Computational Biology, University of Rochester, Rochester, New York.
Physiol Rep. 2014 Sep 11;2(9). doi: 10.14814/phy2.12147. Print 2014 Sep 1.
The in vitro analysis of bacterial-epithelial interactions in the intestine has been hampered by a lack of suitable intestinal epithelium culture systems. Here, we report a new experimental model using an organoid culture system to study pathophysiology of bacterial-epithelial interactions post Salmonella infection. Using crypt-derived mouse intestinal organoids, we were able to visualize the invasiveness of Salmonella and the morphologic changes of the organoids. Importantly, we reported bacteria-induced disruption of epithelial tight junctions in the infected organoids. In addition, we showed the inflammatory responses through activation of the NF-κB pathway in the organoids. Moreover, our western blot, PCR, and immunofluorescence data demonstrated that stem cell markers (Lgr5 and Bmi1) were significantly decreased by Salmonella infection (determined using GFP-labeled Lgr5 organoids). For the first time, we created a model system that recapitulated a number of observations from in vivo studies of the Salmonella-infected intestine, including bacterial invasion, altered tight junctions, inflammatory responses, and decreased stem cells. We have demonstrated that the Salmonella-infected organoid culture system is a new experimental model suitable for studying host-bacterial interactions.
由于缺乏合适的肠道上皮细胞培养系统，肠道中细菌与上皮细胞相互作用的体外分析受到了阻碍。在此，我们报告一种新的实验模型，该模型使用类器官培养系统来研究沙门氏菌感染后细菌与上皮细胞相互作用的病理生理学。利用源自隐窝的小鼠肠道类器官，我们能够观察到沙门氏菌的侵袭性以及类器官的形态变化。重要的是，我们报告了受感染类器官中细菌诱导的上皮紧密连接的破坏。此外，我们展示了通过激活类器官中的NF-κB途径所引发的炎症反应。而且，我们的蛋白质印迹、聚合酶链反应和免疫荧光数据表明，沙门氏菌感染显著降低了干细胞标志物（Lgr5和Bmi1）（使用绿色荧光蛋白标记的Lgr5类器官进行测定）。我们首次创建了一个模型系统，该系统重现了沙门氏菌感染肠道的体内研究中的一些观察结果，包括细菌入侵、紧密连接改变、炎症反应和干细胞减少。我们已经证明，沙门氏菌感染的类器官培养系统是一种适用于研究宿主与细菌相互作用的新实验模型。
