气道黏膜免疫学特征与肺炎支原体肺炎患儿支气管黏液栓塞的关系研究

Relationship between airway mucosal immunological characteristics and bronchial mucus plug in children with Mycoplasma pneumoniae pneumonia

  • 摘要:
    目的 探讨肺炎支原体肺炎(MPP)患儿气道黏膜免疫学特征与支气管黏液栓塞(BMP)的关系。
    方法 选取120例MPP患儿作为研究对象,根据支气管镜检查结果分为有BMP组(检查发现BMP, n=45)和无BMP组(检查未发现BMP, n=75)。采用酶联免疫吸附试验(ELISA)检测支气管肺泡灌洗液(BALF)中细胞因子白细胞介素(IL)-17A、IL-6、IL-8、IL-33、C-X-C基序趋化因子配体10(CXCL10)水平; 采用定量聚合酶链反应(qPCR)检测气道黏蛋白表达(MUC5ACMUC5BMUC1MUC4的mRNA相对表达量)。基于多层螺旋CT影像绘制导航图,对患儿进行支气管镜检查; 通过16S rRNA基因测序分析下呼吸道菌群组成和α-多样性。比较2组患儿住院结局和肺炎复发情况。采用多因素Logistic回归分析筛选BMP发生的影响因素。
    结果 有BMP组患儿BALF中细胞因子(IL-17A、IL-6、IL-8、IL-33和CXCL10)水平均高于无BMP组,差异有统计学意义(P < 0.05)。有BMP组MUC5AC mRNA、MUC5B mRNA表达水平高于无BMP组,差异有统计学意义(P < 0.05); 2组MUC1 mRNA、MUC4 mRNA表达水平比较,差异无统计学意义(P>0.05)。与无BMP组相比,有BMP组患儿在影像学及支气管镜检查中树芽征、支气管充填影及塑型黏液栓等特征更显著,差异有统计学意义(P < 0.05)。有BMP组的微生物α-多样性低于无BMP组,肺炎支原体、流感嗜血杆菌和肺炎链球菌的相对丰度均高于无BMP组,差异有统计学意义(P < 0.05)。有BMP组患儿住院时间长于无BMP组, 90 d第1秒用力呼气容积恢复率低于无BMP组,肺炎复发率和机械通气使用率均高于无BMP组,差异有统计学意义(P < 0.05)。多因素Logistic回归分析显示,树芽征、IL-17A高水平、MUC5AC mRNA高表达和α-多样性降低是BMP发生的有效预测因素(P < 0.05)。
    结论 MPP患儿气道中IL-17A驱动的炎症反应、杯状细胞分泌型黏蛋白MUC5AC过度表达及下呼吸道菌群α-多样性降低,均与BMP的发生密切相关。

     

    Abstract:
    Objective To explore the relationship between airway mucosal immunological characteristics and bronchial mucus plug (BMP) in children with Mycoplasma pneumoniae pneumonia (MPP).
    Methods A total of 120 children with MPP were selected as the study subjects and divided into BMP group (BMP was found during examination, n=45) and non-BMP group (no BMP was found during examination, n=75) according to the results of bronchoscopy. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of cytokines interleukin (IL)-17A, IL-6, IL-8, IL-33, and C-X-C motif chemokine ligand 10 (CXCL10) in bronchoalveolar lavage fluid (BALF). Quantitative polymerase chain reaction (qPCR) was used to detect the expression of airway mucins (the relative mRNA expression levels of MUC5AC, MUC5B, MUC1, and MUC4). Navigation maps were drawn based on multi-slice spiral CT images, and bronchoscopy was performed on the children. The composition and α-diversity of the lower respiratory tract microbiota were analyzed by 16S rRNA gene sequencing. The hospitalization outcomes and pneumonia recurrence of the two groups of children were compared. Multivariate Logistic regression analysis was used to screen the influencing factors for the occurrence of BMP.
    Results The levels of cytokines (IL-17A, IL-6, IL-8, IL-33, and CXCL10) in BALF were higher in the BMP group than those in the non-BMP group, and the differences were statistically significant (P < 0.05). The expression levels of MUC5AC mRNA and MUC5B mRNA were higher in the BMP group than those in the non-BMP group, and the differences were statistically significant (P < 0.05); there were no significant differences in the expression levels of MUC1 mRNA and MUC4 mRNA between the two groups (P>0.05). Compared with the non-BMP group, the features such as tree-in-bud sign, bronchial filling shadow, and plastic mucus plug were more significant in the BMP group during imaging and bronchoscopy, and the differences were statistically significant (P < 0.05). The microbial α-diversity in the BMP group was lower than that in the non-BMP group, and the relative abundances of Mycoplasma pneumoniae, Haemophilus influenzae, and Streptococcus pneumoniae were higher in the BMP group than those in the non-BMP group, with statistically significant differences (P < 0.05). The hospitalization duration of children in the BMP group was longer than that in the non-BMP group, the recovery rate of forced expiratory volume in one second at 90 days was lower than that in the non-BMP group, and the pneumonia recurrence rate and mechanical ventilation usage rate were higher than those in the non-BMP group, with statistically significant differences (P < 0.05). Multivariate Logistic regression analysis showed that tree-in-bud sign, high level of IL-17A, high expression of MUC5AC mRNA, and decreased α-diversity were effective predictive factors for the occurrence of BMP (P < 0.05).
    Conclusion The IL-17A-driven inflammatory response, over-expression of goblet cell secretory mucin MUC5AC, and decreased α-diversity of the lower respiratory tract microbiota in the airways of children with MPP are all closely related to the occurrence of BMP.

     

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