Pediatric Gastroenterology, University of Alberta, Edmonton, Alberta, Canada.
Am J Pathol. 2010 Sep;177(3):1320-32. doi: 10.2353/ajpath.2010.091068. Epub 2010 Jul 22.
Although osteopontin (OPN) is up-regulated in inflammatory bowel diseases, its role in disease pathogenesis remains controversial. The objective of this study was to determine the role of OPN in host responses to a non-invasive bacterial pathogen, Citrobacter rodentium, which serves as a murine infectious model of colitis. OPN gene knockout and wild-type mice were infected orogastrically with either C. rodentium or Luria-Bertani (LB) broth. Mouse-derived OPN(+/+) and OPN(-/-) fibroblasts were incubated with C. rodentium and attaching-effacing lesions were demonstrated using transmission electron microscopy and immunofluorescence. Colonic expression of OPN was increased by C. rodentium infection of wild-type mice. Furthermore, colonic epithelial cell hyperplasia, the hallmark of C. rodentium infection, was reduced in OPN(-/-) mice, and spleen enlargement by infection was absent in OPN(-/-) mice. Rectal administration of OPN to OPN(-/-) mice restored these effects. There was an 8- to 17-fold reduction in bacterial colonization in OPN(-/-) mice, compared with wild-type mice, which was accompanied by reduced attaching-effacing lesions, both in infected OPN(-/-) mice and OPN(-/-) mouse fibroblasts. Moreover, adhesion pedestals were restored in OPN(-/-) cells complemented with human OPN. Therefore, lack of OPN results in decreased pedestal formation, colonization, and colonic epithelial cell hyperplasia responses to C. rodentium infection, indicating that OPN impacts disease pathogenesis through bacterial attachment and altered host immune responses.
虽然骨桥蛋白 (OPN) 在炎症性肠病中上调,但它在疾病发病机制中的作用仍存在争议。本研究的目的是确定 OPN 在宿主对非侵入性细菌病原体柠檬酸杆菌的反应中的作用,柠檬酸杆菌是结肠炎的小鼠感染模型。通过口胃感染或罗氏肉汤感染 OPN 基因敲除和野生型小鼠。用柠檬酸杆菌孵育源自小鼠的 OPN(+/+)和 OPN(-/-)成纤维细胞,并通过透射电子显微镜和免疫荧光证明附着破坏病变。野生型小鼠感染柠檬酸杆菌可增加结肠中 OPN 的表达。此外,OPN(-/-)小鼠的结肠上皮细胞过度增生减少,这是柠檬酸杆菌感染的标志,而 OPN(-/-)小鼠感染时脾脏肿大不存在。OPN(-/-)小鼠直肠给予 OPN 可恢复这些作用。与野生型小鼠相比,OPN(-/-)小鼠的细菌定植减少了 8 到 17 倍,这伴随着附着破坏病变的减少,无论是在感染的 OPN(-/-)小鼠还是 OPN(-/-)小鼠成纤维细胞中都是如此。此外,在补充人 OPN 的 OPN(-/-)细胞中恢复了粘附基脚。因此,缺乏 OPN 导致附着基脚形成、定植和结肠上皮细胞增生对柠檬酸杆菌感染的反应减少,表明 OPN 通过细菌附着和改变宿主免疫反应影响疾病发病机制。