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目的矽肺是一种以肺纤维化为病理特征的疾病,其发病机制仍不完全清楚,外周血单核细胞表型偏移失衡在肺纤维化病理进展中发挥着重要作用。文中旨在研究石英诱导小鼠矽肺(pneumosilicosis)模型中循环单核细胞亚群在不同病理阶段的变化特点,探讨其动态变化与肺炎症损伤和纤维化的关系。方法 100只雄性C57BL/6小鼠(18~22 g)随机数字表法分为等渗盐水组和石英组,石英组滴注吸入40 m L石英混悬液(100 mg/kg),等渗盐水组滴注吸入相同体积无菌等渗盐水。经术后第1、3、7、14和28天应用流式细胞术检测循环单核细胞亚群变化;常规支气管肺泡灌洗术后进行炎症细胞分类计数;HE染色和苦味酸天狼星红染色检测小鼠肺组织炎症评分及胶原容积分数。结果病理染色显示,造模后第7天,小鼠肺组织可见明显的矽结节;与等渗盐水组相比,石英组小鼠肺组织炎症评分、胶原容积分数显著升高[(1.400±0.089)vs(0.920±0.049)、(1.950±0.065)vs(0.525±0.048),P<0.01],并持续到第28天[(1.520±0.136)vs(0.800±0.089),(5.300±0.776)vs(0.850±0.050),P<0.01];与相应时间点等渗盐水组比较,石英组BAFL细胞总数于第1天开始升高[(7.693±2.495)vs(2.008±0.901),P<0.01],在第3天达到高峰[(13.346±3.563)vs(1.044±0.695)],随后第7、14、28天呈现递减趋势,但仍都高于等渗盐水组(P<0.01);石英组BALF巨噬细胞绝对值在第3天时显著高于等渗盐水组(6.821±2.627 vs 0.980±0.663,P<0.01),并在第7天维持于高水平状态[(6.697±1.864)vs(1.225±0.601),P<0.01],后呈下降趋势,但第14及28天巨噬细胞绝对值仍高于等渗盐水组(P<0.01);石英组BALF中性粒细胞绝对值在第1、3、7及14天都显著高于等渗盐水组(P<0.01),且呈现出逐渐下降的趋势,在第28天与等渗盐水组差异无统计学意义(P>0.05)。石英组小鼠Ly6Chi单核细胞亚群占比在所有检测时间点均显著高于等渗盐水组(P<0.01),其中第7天达到高峰[(78.300±2.517)vs(58.750±2.386),P<0.01];Ly6Clo亚群比例变化与之相反。第7天和第28天的小鼠Ly6Chi单核细胞亚群占比与相应时间点的IS及CVF值呈正相关(P<0.01)。结论循环Ly6Chi和Ly6Clo单核细胞亚群占比在石英诱导的小鼠矽肺的不同病理时期呈现动态变化,持续性的Ly6Chi单核细胞亚群占比升高可能与矽肺的急慢性炎症及后期的纤维化程度存在密切的联系。
The purpose of silicosis is a pathological feature of pulmonary fibrosis disease, the pathogenesis is still not fully understood, peripheral blood monocyte phenotype offset imbalance in pathological progress of pulmonary fibrosis plays an important role. The aim of this study was to investigate the changes of circulating monocyte subsets in quartz-induced mouse pneumosilicosis at different pathological stages and to explore the relationship between the dynamic changes and lung injury and fibrosis. Methods A total of 100 male C57BL / 6 mice (18-22 g) were randomly divided into isotonic saline group and quartz group. Quartz group was infused with 40 mL quartz suspension (100 mg / kg) Saline group instillation inhaled the same volume of sterile isotonic saline. The changes of the circulating monocyte subsets were detected by flow cytometry on the 1st, 3rd, 7th, 14th and 28th day postoperatively. The inflammatory cells were sorted and counted after routine bronchoalveolar lavage (HE staining) and picric acid Sirius red staining Mouse lung tissue inflammation score and collagen volume fraction. Results The pathological staining showed that on the 7th day after the model establishment, the obvious pulmonary nodules were observed in the lung tissue of mice. Compared with the isotonic saline group, the lung tissue inflammatory score and the collagen volume fraction in the quartz group mice were significantly increased [(1.400 ± (1.520 ± 0.136) vs (0.800 ± 0.089), (5.300 ± 0.776) vs (0.920 ± 0.049), (1.950 ± 0.065) vs (0.525 ± 0.048), P <0.01] vs (0.850 ± 0.050), P <0.01]. Compared with the isotonic saline group at the corresponding time point, the total number of BAFL cells in quartz group increased from the first day [(7.693 ± 2.495) vs (2.008 ± 0.901) P <0.01 ] Peaked on day 3 [(13.346 ± 3.563 vs 1.044 ± 0.695)], and then decreased on the 7th, 14th and 28th days, but still higher than that of the isotonic saline group (P <0.01) The absolute value of BALF macrophages in group 3 was significantly higher than that in isotonic saline group on the 3rd day (6.821 ± 2.627 vs 0.980 ± 0.663, P <0.01), and maintained at a high level on the 7th day [(6.697 ± 1.864 vs vs 1.225 ± 0.601), P <0.01], and then decreased, but the absolute values of macrophages on the 14th and 28th day were still higher than that of the isotonic saline group (P <0.01) 1,3,7 and 14 days were significantly higher than the isotonic saline group (P <0.01), and showed a gradual Descending trend, the difference in isotonic saline group 28 days no significant (P> 0.05). The proportion of Ly6Chi monocyte subsets in the quartz group was significantly higher than that in the isotonic saline group at all time points (P <0.01), with the peak on the 7th day (78.300 ± 2.517 vs 58.750 ± 2.386, P <0.01]. The proportion of Ly6Clo subpopulations changed in the opposite direction. The proportion of Ly6Chi monocyte subsets on day 7 and day 28 was positively correlated with IS and CVF at the corresponding time points (P <0.01). Conclusions The proportion of circulating Ly6Chi and Ly6Clo monocyte subsets shows a dynamic change in silicosis-induced mouse silicosis at different pathological stages. The increased proportion of persistent Ly6Chi monocyte subsets may be related to the acute and chronic inflammation of silicosis and the late The degree of fibrosis is closely linked.