肺结节病患者支气管肺泡灌洗液中细胞外 microRNA 的表达分析。

Expression analysis of extracellular microRNA in bronchoalveolar lavage fluid from patients with pulmonary sarcoidosis.

机构信息

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.

Department of Respiratory Medicine and Tuberculosis, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.

出版信息

Respirology. 2018 Dec;23(12):1166-1172. doi: 10.1111/resp.13364. Epub 2018 Jun 29.

DOI:10.1111/resp.13364

PMID:29956871

Abstract

BACKGROUND AND OBJECTIVE

MicroRNA (miRNA) are transcriptional regulators implicated in pulmonary sarcoidosis and packaged in extracellular vesicles (EV) during cellular communication. We characterized EV and investigated miRNA expression in bronchoalveolar lavage (BAL) fluid from sarcoidosis patients.

METHODS

EV were characterized for size(s) using dynamic light scattering and transmission electron microscopy (TEM) analysis and protein markers by immunoblotting. Twelve extracellular and 5 cellular miRNA were investigated in BAL from 16 chest X-ray stage-I (CXR-I) and 17 CXR stage-II (CXR-II) sarcoidosis patients. Associations between miRNA and disease characteristics (extrapulmonary involvement, pulmonary function and BAL cell profile) were statistically analysed.

RESULTS

BAL from sarcoidosis patients contained exosomes and microvesicles (MV) as EV. In these EV, expression of miR-146a (P = 0.007), miR-150 (P = 0.003) and BAL cellular miR-21 (P = 0.01) was increased in CXR-II compared with CXR-I. Other detected EV (miR-21 and miR-26a) and cellular (miR-31, miR-129-3p, miR-146a and miR-452) miRNA were not differentially expressed. The investigated miRNA did not reflect extrapulmonary involvement, but EV miR-146a and miR-150 were negatively correlated with pulmonary function (miR-146a with vital capacity (VC; Spearman's correlation coefficient (r ), P = -0.657, 0.007), percent predicted forced expiratory volume in 1 s (FEV ; -0.662, 0.006) and FEV /forced vital capacity (FVC) ratio (-0.649, 0.008); miR-150 correlated negatively with VC (-0.584, 0.019) and FEV /FVC ratio (-0.746, 0.001) in CXR-II cases).

CONCLUSION

Our data provide evidence that exosomes and microvesicles as extracellular vesicles are present in the bronchoalveolar space of sarcoidosis patients and they differentially express EV miRNA (miR-146a and miR-150), the expression of which correlates negatively with pulmonary function indices. The significance of these findings for disease pathophysiology and clinical course require further investigation.

摘要

背景与目的

微小 RNA（miRNA）是参与肺结节病的转录调控因子，并在细胞间通讯过程中被包裹在细胞外囊泡（EV）中。我们对 EV 进行了特征描述，并研究了结节病患者支气管肺泡灌洗液（BAL）中的 miRNA 表达。

方法

使用动态光散射和透射电子显微镜（TEM）分析对 EV 的大小进行了特征描述，并通过免疫印迹法对其蛋白质标志物进行了研究。我们研究了来自 16 名胸部 X 射线分期 I（CXR-I）和 17 名 CXR 分期 II（CXR-II）结节病患者 BAL 中的 12 种细胞外和 5 种细胞内 miRNA。使用统计学方法分析了 miRNA 与疾病特征（肺外受累、肺功能和 BAL 细胞特征）之间的相关性。

结果

结节病患者的 BAL 中含有外泌体和微泡（MV）等 EV。与 CXR-I 相比，CXR-II 患者的 EV 中 miR-146a（P=0.007）、miR-150（P=0.003）和 BAL 细胞中的 miR-21（P=0.01）表达增加。其他检测到的 EV（miR-21 和 miR-26a）和细胞内 miRNA（miR-31、miR-129-3p、miR-146a 和 miR-452）则没有差异表达。所研究的 miRNA 并未反映肺外受累情况，但 EV miR-146a 和 miR-150 与肺功能呈负相关（miR-146a 与肺活量（VC；Spearman 相关系数（r），P=0.657，0.007）、1 秒用力呼气量（FEV）的预计百分比（-0.662，0.006）和 FEV/用力肺活量（FVC）比值（-0.649，0.008）；miR-150 与 CXR-II 病例中的 VC（-0.584，0.019）和 FEV/FVC 比值（-0.746，0.001）呈负相关）。