PATHOPHYSIOLOGICAL ANALYSIS OF BRONCHIOLITIS OBLITERANCE USING CD40-DEFICIENT MICE AND GFP-TRANSGENIC MICE

据报道，缩窄性闭塞性毛细支气管炎(BO)患者涉及骨髓移植和心肺移植，以及使用或不使用青霉胺治疗的类风湿性关节炎患者。虽然缩窄性BO是一种相对罕见的疾病，但由于骨髓和心肺移植等同种异体移植的数量不断增加，它最近已成为人们重新关注的焦点。为了探讨BO的发病机制，我们建立了BO的小鼠实验模型，并对CD40分子和骨髓来源的祖细胞的作用进行了研究。在此基础上，我们进一步研究了仙人掌诱发的BO。给野生型(WT)和CD40KO小鼠气管内注射内毒素后，CD40KO小鼠的肺损伤明显减轻。此外，内毒素诱导的诱导型一氧化氮合酶(INOS)表达和一氧化氮(NO)在CD40KO小鼠肺中的表达也受到抑制。此外，测试…CD40KO组小鼠肺泡灌洗液中炎症介质α、IL-1b、巨噬细胞炎性蛋白-2、活性氧、氮中间产物和基质金属蛋白酶-9的释放明显减少。我们在体外研究了小鼠肺泡巨噬细胞(AMF)的功能。CD40KO AMF中未检测到iNOS的表达，而WT AMF中有iNOS的表达。此外，我们使用GFP骨髓嵌合体小鼠检测了骨髓来源的祖细胞在博莱霉素诱导的肺部炎症中的作用。诱导肺部炎症导致GFP+细胞增多，积聚到活动性纤维化病变中。GFP+细胞也表达I型胶原。此外，我们还用引起BO的叶灌木Sauropus androynus刺激了U937单核细胞来源的细胞系。我们发现了肿瘤坏死因子-α的产生，但没有发现CXCL9或CXCL10的产生。提示肿瘤坏死因子-α可能是BO发病的重要因素之一。我们的数据表明，CD40或骨髓祖细胞的功能阻断将成为包括BO在内的肺损伤临床治疗的靶点之一。较少

Cases of constrictive bronchiolitis obliterans(BO) have been reported involving patients with bone marrow transplants and heart/lung transplants as well as those with rheumatoid arthritis with or without penicillamine treatment. Although constrictive BO was a relatively rare disease, it has recently become the focus of renewed interest because the number of allograft recipients such as bone marrow and heart/lung transplants has been increasing. To investigate the pathogenesis of BO, we focused on the establishment of mouse experimental mode for BO and the role of CD40 molecule and the bone marrow-derived progenitor cells. Further, we investigate the Sauropus Androgynus-induced BO. When wild-type(WT) mice and CD40KO mice were injected intratracheally with LPS, LPS-induced lung injury was significantly reduced in CD40KO mice. Further, LPS-induced inducible nitric oxide synthase(iNOS) expression and nitric oxide(NO) production was also inhibited in the lungs of CD40KO mice. In addition, t … More he release of inflammatory mediators, that is, TNF-α,IL-1b, macrophage inflammatory protein 2(MIP-2), reactive oxygen, nitrogen intermediates and MMP-9 into the bronchoalveolar lavage fluid, was significantly reduced in CD40KO mice. We studied the function of alveolar macrophages(AMf) in each of mice ex vivo. Although iNOS in WT AMf was induced in response to LPS, no iNOS expression could be detected in CD40KO AMf. In addition, we examined the role of bone marrow-derived progenitor cells in bleomycin-induced pulmonary inflammation using GFP bone marrow chimera mice. Induction of pulmonary inflammation resulted in the increase of GFP+ cells accumulated into the active fibrotic lesions. GFP+ cells also expressed type I collagen. Further, we stimulated the monocyte derived cell lines of U937 with Sauropus Androgynus, which is a leaf shrub for the cause of BO. We found the production of TNF-α, but not CXCL9 or CXCL10. These results indicated that TNF-α might be one of important factor for the pathogenesis of BO. Our data suggest that the functional blockade of CD40 or bone marrow-derived progenitor cells would yield one of the targets for the clinical treatment for lung injury including BO. Less