慢性阻塞性肺疾病急性加重患者中副流感嗜血杆菌基因型分布及对气道上皮细胞的影响

目的研究慢性阻塞性肺疾病急性加重(AECOPD)患者中不同基因型副流感嗜血杆菌(HPI)的分布及其对气道上皮细胞株A549的影响。方法以2010年至2011年间中山大学附属第三医院住院的AECOPD患者为研究对象,采集痰标本培养HPI,所得菌株采用随机引物聚合酶链反应(RAPD)进行基因分型,并与气道上皮细胞株A549共同培养,作用不同时间后取细胞培养上清行白细胞介素6(IL-6)、IL-8的检测,并通过显微镜观察细胞形态变化。结果入组80例AECOPD患者,按症状分为3型,共培养HPI 15株,培养阳性率18.8%。1型与3型两组间细菌阳性率比较有显著差异。所得菌株与A549上皮细胞共培养后IL-8、IL-6分泌水平均较对照组明显升高,且与时间呈正相关。所得菌株采用RAPD法可分为4种基因型,RAPDⅢ型与A549上皮细胞共培养后,12 h和24 h的IL-8分泌水平均高出其他组。各组经显微镜观察均未见明显细胞形态学变化,且HPI未进入细胞。结论不同临床类型AECOPD患者痰HPI培养阳性率不同。HPI无法进入气道上皮细胞胞内,可能是胞外感染菌,但能够引起气道上皮细胞炎症因子表达持续升高,而且不同基因型毒力不同。HPI可能是COPD急性加重的致病菌之一。

Genotype Distribution of HaemophUus Parainfluenzae from Patients with Acute Exacerbations ofChronic Obstructive Pulmonary Disease and Their Effects on A549 Cells

Objective To observe the genotype distribution of Haemophilus parainfluenzae from patients with acute exacerbations of chronic obstructive pulmonary disease （AECOPD） and their effects on A549 cells. Methods 80 hospitalized patients with AECOPD in our hospital were enrolled. Haemophilus parainfluezae were collected by sputum culture and genotyped,then inoculated with cell line A549. IL-6 and IL-8 concentrations in the supernatant were detected and cell morphology was observed at different time points. Results The patients were divided into three groups according to their symptoms. 15 Haemophilus parainfuenzae strains were collected and the positive culture rate between type 1 and type 3 COPD patients were statistically different. The concentrations of IL-6 and IL-8 were both significantly higher than control and increased as time passed. 4 genotypes were got by random amplification of polymorphic DNA （RAPD）. In RAPD Ⅲ group,the IL-8 concentration was higher at 12h and 24h than others. No morphotogic change was found in the cells inoculated with Haemophilus parainfuenzae by microscope after fixing. Conclusions Positive culture rate of Haemophilus parainfuenzae was different in different COPD groups according to symptoms. Haemophilus parainfuenzae can stimulate a cytokine response in A549 cells, maybe one of the pathogens of AECOPD, especially the RAPD III type. Haemophilus parainfuenzae is not an intracellular bacteria.