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目的探讨血管内皮细胞间紧密连接的改变在肺纤维化发生发展过程中的作用。方法选用SD大鼠72只,随机分为博莱霉素(BLM)组、肺炎组及正常对照组。分别采用BLM、肺炎双球菌及生理盐水气管内1次灌注的方法制作大鼠肺损伤模型及正常对照模型,分别于第3、7、14、28天取动物肺组织制作成电镜标本,观察肺毛细血管内皮细胞间紧密连接的变化,并运用免疫组织化学方法观察单核巨噬细胞在肺间质中的浸润情况。结果光镜:肺炎第3天组大鼠肺泡壁水肿,肺组织内大量中性粒细胞浸润;第3天及第7天组大鼠局部肺实变,肺组织内巨噬细胞浸润增加,其阳性面积比与正常对照组比较差异有统计学意义(P<0.05);第14天组肺间质内有少量巨噬细胞浸润;第28天组肺泡壁结构完整,未见明显炎细胞浸润,巨噬细胞阳性面积比与正常对照组比较差异无统计学意义(P>0.05)。BLM第3天及第7天组大鼠肺组织实变、出血,其中炎细胞浸润情况与同期肺炎组类似;第14天及第28天组大鼠肺间质内持续存在单核巨噬细胞浸润,其阳性面积比与正常对照组比较差异有统计学意义(P<0.05)。此阶段同时伴有肺泡壁增厚,间质内胶原纤维沉积。电镜:肺炎组大鼠肺毛细血管内皮细胞间紧密连接在第3天及第7天组时完全开放,镧颗粒通过细胞连接并沉积在基膜上,第28天组大部分细胞间连接紧密,与正常对照组比较无明显差异。BLM组第3、7、14天及28天组,大部分内皮细胞紧密连接处于开放状态。结论博莱霉素鼠肺毛细血管内皮细胞间紧密连接持续开放,与巨噬细胞在组织间浸润密切相关,并可能是博莱霉素致肺纤维化发生的重要因素。
Objective To investigate the role of vascular endothelial cell junctional changes in the development of pulmonary fibrosis. Methods 72 SD rats were randomly divided into BLM group, pneumonia group and normal control group. The models of lung injury and normal control were established by intraperitoneal instillation of BLM, Streptococcus pneumoniae and saline respectively. The lungs of animals were made into electron microscopy specimens on days 3, 7, 14 and 28, respectively. Capillary endothelial cells tightly connected changes, and the use of immunohistochemistry to observe the mononuclear macrophages in the interstitial infiltration of the lungs. Results Light microscopy: Alveolar wall edema in rats on day 3 of pneumonia and massive neutrophil infiltration in lung tissue. On the 3rd and 7th day, local lung consolidation and macrophage infiltration in lung tissue increased (P <0.05). There was a small amount of macrophage infiltration in the interstitium on the 14th day. On the 28th day, the alveolar wall was intact with no obvious inflammatory cell infiltration, The positive area ratio of macrophages was not significantly different from the normal control group (P> 0.05). On the 3rd and 7th day after BLM, the lung tissue of the rats in the BLM group were solidified and hemorrhaged, and the infiltration of inflammatory cells was similar to that in the pneumonia group. On the 14th and 28th days, the mononuclear macrophages Infiltration, the positive area ratio compared with the normal control group, the difference was statistically significant (P <0.05). This stage accompanied by alveolar wall thickening, interstitial collagen deposition. Electron microscopy: Pneumococcal pulmonary capillary endothelial cells in pneumonia rats were completely open on day 3 and 7, and lanthanum particles were connected to and deposited on the basilar membrane via the cells. Most of the cells in group P were closely connected to each other on day 28, No significant difference compared with the normal control group. On the 3rd, 7th, 14th day and the 28th day in the BLM group, most of the endothelial cells were tight junctions in the open state. Conclusion Bleomycin-induced pulmonary capillary endothelial cells tight junctions continue to open, and macrophages in the inter-organizational infiltration is closely related to bleomycin-induced pulmonary fibrosis may be an important factor.