不同临床表型COPD频繁和非频繁发作患者气道巨噬细胞的异质性

目的：比较频繁发作(frequent exacerbator，FE)和非频繁发作(infrequent exacerbator，iFE)的不同临床表型的 慢性阻塞性肺病(chronic obstructive pulmonary disease，COPD)患者的临床特征及诱导痰中巨噬细胞的异质性。方法： 分析80例慢性支气管炎(chronic bronchitis，CB)、肺气肿(emphysema，EM)、哮喘-COPD重叠(asthma-COPD overlap， ACO)表型的COPD急性发作住院患者的临床特征；采用流式细胞术检测诱导痰巨噬细胞CCL3和CD163的表达，定 量聚合酶链反应(quantitative PCR，qPCR)检测HIF-1α和Cav-1的表达。结果：FE和iFE患者在年龄，第1 s用力呼气容积 (forced expiratory volume in one second，FEV1)/用力肺活量(forced vital capacity，FVC)，痰细菌阳性率，COPD评估测试 (COPD Assessment Test，CAT)评分，改良医学研究委员会(Modifi ed Medical Research Council，mMRC)评分方面的差异 有统计学意义(P<0.05)；相对于iFE患者，FE患者诱导痰巨噬细胞上CCL3荧光强度明显降低(P<0.01)，而CD163显著增 高(P<0.01)，同时HIF-1α和Cav-1 mRNA水平也显著升高(P<0.01)。CB表型的FE患者与iFE患者之间在年龄，痰细菌阳 性率，CAT评分，mMRC评分方面的差异有统计学意义(P<0.05)；相对于iFE患者，FE患者诱导痰巨噬细胞上CCL3荧 光强度轻度降低(P>0.05)，而CD163显著增高(P<0.01)，同时HIF-1α和Cav-1 mRNA水平也显著升高(P<0.01)。EM表型 的FE患者与iFE患者之间在年龄，病程，FEV1/FVC，痰细菌阳性率，CAT评分，mMRC评分方面的差异有统计学意义 (P<0.05)；相对于iFE患者，FE患者诱导痰巨噬细胞CCL3荧光强度轻度减低(P>0.05)，而CD163轻度升高(P>0.05)，同 时HIF-1α水平轻度升高(P>0.05)，Cav-1水平则显著增加(P<0.01)。ACO表型的FE与iFE患者所有临床特征差异均无统计 学意义，FE患者诱导痰巨噬细胞CCL3荧光强度明显低于iFE患者(P<0.01)，而CD163差异无统计学意义(P>0.05)，同 时HIF-1α(P<0.01)和Cav-1(P<0.05)表达也显著增加。全部FE及CB表型FE患者CCL3与HIF-1α和Cav-1均呈显著负相关， CD163仅与HIF-1α呈显著正相关；EM表型FE患者CD163与HIF-1α呈显著正相关；ACO表型FE患者CCL3与HIF-1α呈显 著负相关，而CD163与HIF-1α呈显著正相关。结论：CB，EM，ACO表型FE和iFE患者临床特征差异不一，诱导痰中替 代活化巨噬细胞(M2)在FE患者中占优势，HIF-1α可能在其极化过程中起关键作用。

Heterogeneity of airway macrophage in different clinical phenotypes of COPD patients with frequent or infrequent exacerbations

Objective: To compare the clinical features and the heterogeneity of macrophages in different clinical phenotypes of chronic obstructive pulmonary disease (COPD) patients with frequent or infrequent exacerbations. Methods: Clinical characteristics of eighty COPD patients with chronic bronchitis (CB), emphysema (EM) or asthma-COPD overlap (ACO) phenotypes suffered from acute exacerbation were analyzed. The expressions of CCL3 and CD163 in sputum macrophages were detected by flow cytometry. The expressions of HIF-1α and Cav-1 in sputum macrophages were detected by quantitative PCR (qPCR). Results: The age, forced expiratory volume in one second (FEV1)/forced vital capacity (FVC), sputum bacteria positive rate, COPD Assessment Test (CAT) score, and Modified Medical Research Council (mMRC) score between the patients with FE and iFE were significantly different (P<0.05). Compared with iFE patients, the fluorescence intensity of CCL3 in sputum macrophages from patients with FE was significantly lower (P<0.01), while CD163 was significantly increased (P<0.01). Meanwhile, HIF-1α and Cav-1 mRNA levels were also significantly increased (P<0.01). The age, sputum bacteria positive rate, CAT score, and mMRC score between the patients of FE and iFE with CB phenotype were significantly different (P<0.05). Compared with iFE patients, the fluorescence intensity of CCL3 in sputum macrophages from FE patients was slightly decreased (P<0.05), while CD163 was significantly raised (P<0.01). Meanwhile, HIF-1α and Cav-1 mRNA levels were also significantly increased (P<0.01). The age, duration of disease, FEV1/FVC, sputum bacteria positive rate, CAT score, and mMRC score between the patients of FE and iFE with EM phenotype were significantly different (P<0.05). Compared with iFE patients, the fluorescence intensity of CCL3 in sputum macrophages from FE patients was slightly decreased (P>0.05), while CD163 was slightly raised (P>0.05). Meanwhile, HIF-1α levels were slightly elevated (P>0.05), while Cav-1 expression was significantly increased (P<0.01). There were no significant differences in all clinical features between FE and iFE patients with ACO phenotype. The fluorescence intensity of CCL3 in sputum macrophages from patients with FE was significantly lower than that in iFE patients (P<0.01); there was no significant difference in CD163 (P>0.05). At the same time, the expression of HIF-1α (P<0.01) and Cav-1(P<0.05) also increased significantly. There was a significant negative correlation between CCL3 and HIF-1α or Cav-1 in all FE and FE patients with CB phenotype. CD163 was only positively correlated with HIF-1α in those patients and FE patients with EM phenotype. There was a significant negative correlation between CCL3 and HIF-1α in FE patients with ACO phenotype, while CD163 was significantly positively correlated with HIF-1α. Conclusion: The clinical features of FE or iFE patients with CB, EM or ACO phenotype are different, and M2 in induced sputum from FE patients are dominant. HIF-1α may play a key role in the polarization process.