肝肺综合征大鼠模型肺组织巨噬细胞表型变化

目的　探讨复合因素诱导大鼠肝肺综合征（hepatopulmonary syndrome，HPS）发病过程中巨噬细胞的表型变化特点。 方法　制备肝硬化合并HPS大鼠模型，随机设立4周组、6周组、8周组3个时点，同期设置正常对照组。采用HE染色观察肝、肺组织病理变化；VG染色法对肝纤维化进行观察；免疫荧光法对巨噬细胞标志物CD68进行观察，RT-PCR检测肺组织标本中M1、M2巨噬细胞表型标记物CD86及CD206，Elisa检测iNOS、TNF-α、Arg-1及IL10。 结果　随病程进展，模型组大鼠CD86、iNOS及TNF-α在肺组织中表达逐渐增高，CD86及iNOS至6周达到高峰后有所回落，但仍明显高于正常对照组；CD206、Arg-1及IL-10的表达于第6周开始明显增加，第8周CD206及Arg-1亦有回落。上述指标在各时点的表达均有显著差异；其中，模型组大鼠肺组织iNOS与CD86呈正相关；Arg-1与CD206呈正相关；IL-10与CD206呈正相关。 结论　HPS的发病过程中，肺组织巨噬细胞经历了一系列的表型变化，6周前以M1为主，第8周以M2为主。

Phenotypic change of macrophages in lung tissue during the development of hepatopulmonary syndrome

Objective　To investigate the phenotypic change of macrophages in lung tissue during the development of hepatopulmonary syndrome (HPS).　Methods　The HPS model was induced by multiple pathogenic factors in Sprague-Dawley rats and divided into HPS groups of the 4th week，the 6th week and the 8th week. Normal control groups at the corresponding time points were also set up. Liver and Lung tissue histology was observed by HE staining. VG staining method was used to observe the liver fibrosis. Immunofluorescence method was used to observe the macrophage marker (CD68). The expressions of M1/M2 macrophage markers (CD86 and CD206) were detected by real-time PCR assay, while the expressions of corresponding secretions iNOS, TNF-α, Arg-1 and IL10 were detected by Elisa.　Results　With the progress of the disease, the expression of CD86, iNOS and TNF-α gradually increased in the lung tissue of model group. After reaching the peak in the 6th weeks, CD86 and iNOS fell back. While the expression of CD206, Arg-1 and IL10 began to increase significantly at the 6th week. There was a positive correlation between iNOS and CD86 in the lung tissue of the model group. Arg-1 was positively correlated with CD206. IL10 was positively correlated with CD206.　Conclusions　During the pathogenesis of HPS, macrophages in lung tissue underwent a series of phenotypic changes, with M1 predominating at 6 weeks and M2 predominating at 8 weeks.