慢性阻塞性肺疾病患者和吸烟者CD14＋单核细胞Toll样受体的表达及意义

Expression of Toll.like receptors on CD14 ＋ monocytes from patients with chronic obstructive pulmonary disease and smokers

目的探讨稳定期慢性阻塞性肺疾病（COPD）患者和肺功能正常的吸烟者外周血CD14＋单核细胞Toll样受体（TLR）表达的变化及意义。方法选择COPD稳定期患者30例（COPD组）、健康吸烟者21例（吸烟组）及健康不吸烟者25例（不吸烟组），用肺功能仪测定第一秒用力呼气容积（FEV1）占预计值的百分比（FEV1占预计值％）和FEV1／用力肺活量（FVC）；应用流式细胞仪全血双色免疫荧光直标法检测外周血CD14＋单核细胞的TLR2、TLR4表达水平，结果以CD14＋细胞TLR相对平均荧光强度（rmfi）和TLR相对阳性细胞百分率（rpcp）表示，并进行TLR2、TLR4表达与肺功能指标相关性分析。结果COPD组CD14＋单核细胞TLR2表达的rmfi（6．3±1．4）和rpcp（52．9％±20．5％）均明显低于吸烟组（分别为8．2±2．2和73．5％±19．0％，均P〈0．01）和不吸烟组（分别为11．0±2．4和82．8％±17．9％，均P〈0．01）；TLR4表达的rmfi（2．2±0．9）明显低于不吸烟者组（3．0±0．5，P〈0．01），与吸烟组（2．5±0．6）差异无统计学意义（P〉0．05），而rpcp（M=1．3％，Q1-Q3为0．7％～2．4％）则明显低于吸烟组（M=4．7％，Q1～Q3为2．7％～9．4％，P〈0．01）和不吸烟组（M=5．3％，Q1～Q3为2．6％-8．4％，P〈0．01）。吸烟组CD14＋单核细胞中TLR2、TLR4表达的rmfi均明显低于不吸烟组（P〈0．01，P〈0．05）；而rpcp与不吸烟组差异无统计学意义（P〉0．05）。CD14＋单核细胞TLR2和TLR4的表达与FEV1占预计值％、FEV1／FVC呈显著正相关（均P〈0．01）。结论稳定期COPD患者和吸烟者外周血CD14＋单核细胞TLR2及TLR4表达显著下调，固有免疫可能受到抑制，TLR表达下调与肺功能下降有关。

Objective To investigate the expression of toll-like receptor-2 （TLR2）, TLR4 on the CD14 ＋ monocytes of patients with stable chronic obstructive pulmonary disease （COPD） and healthy smokers, and to explore the role of TLR2 and TLR4 in COPD pathogenesis. Methods Thirty COPD patients without evidence of acute exacerbation, 21 healthy smokers, and 25 healthy non-smokers underwent measurement of forced expiratory volume in 1 second （FEV1） % predicted and FEVJ forced vital capacity （FVC） by spirometry. The expression of TLR2 and TLR4 surface molecules on human CD14 ＋ monocytes was assessed using fluorescence activated cell sorter analysis by flow cytometry, expressed as relative mean fluorescence intensity （rmfi） and relative positive cell percent （rpcp）, and the correlation of TLR expression with lung function parameters was analyzed. Results The rmfl and rpcp of TLR2 on CD14 ＋ monocytes of the COPD patients were （6.3 ± 1.4） % and （52.9 ± 20.5） % respectively, both significantly lower than those of the healthy smokers C （8.2 ±2.2 ） % and （73.5 ± 19.0） % respectively ] and those of the nonsmokers [ （ 11.0 ± 2.4 ） % and （82.8 ± 17.9 ） % respectively, all P 〈 0.01 ） ]. The rmfl of TLR4 of the COPD patients was 2.2 ±0.9, significantly lower than that of the nonsmokers （3.0 ±0.5, P 〈0.01 ） , while similar to that of the healthy smokers （2.5 ± 0.6, P 〉 0.05 ）. The rpcp of TLR4 of the COPD patients （M=1.3%,QL - Q3： 0.7% -2.4%） was significantly lower than that of the healthy smokers （M= 4.7%,Q1-Q3： 2.7%-9.4%, P〈0.01） and nonsmokers （M=5.3%, Q1 -Q3： 2.6% -8.4%, P〈 0.01 ）. The rmfi of TLR2 and TLR4 on CD14 ＋ monocytes of the healthy smokers was lower than that of the nonsmokers （P 〈 0.05 ） , while the rpcp of TLR2 and TLR4 on CD14 ＋ monocytes of the healthy smokers was similar to that of the nonsmoker （ P 〈 0. 05 ）. The expression of TLR2 and TLR4 on monocytes was positively correlated with the lung function parameters, including FEV1% predicted and FEV1/FVC （ all P〈0.01）.Conehmlons The expression levels of TLR2 and TLR4 on CD14 ＋ monocytes of the stable COPD patients and healthy smokers decreased significantly. The innate immune response may be depressed in the COPD patients and smokers, and the down-regulation of TLR is associated with reduced lung function parameters.