慢性阻塞性肺病外周血单个核细胞中的组蛋白去乙酰化酶活性与全身炎症的关系

Relationship Between Systemic Inflammation and Activity of Histone Deacetylase in Peripheral Blood Mononuclear Cells from COPD Patients

目的:观察慢性阻塞性肺病(COPD)患者外周血单个核细胞(PBMC)组蛋白去乙酰化酶(HDAC)的活性及其与全身炎症的关系。方法:募集COPD患者及吸烟和非吸烟健康志愿者并检测其肺功能。抽取所有被研究者外周静脉血,以密度梯度离心法分离出外周血单个核细胞。提取组蛋白并测定HDAC活性。测定所有被研究者外周血清的IL-8及IL-10水平,并观察其与HDAC的关系。结果:外周血单个核细胞中HDAC活性与外周血清中IL-8水平呈负相关(在COPD患者中,r=-0.693,P=0.000;在所有被研究者中,r=-0.598,P=0.000)。外周血IL-8水平在健康吸烟者高于健康不吸烟者2倍以上[(106.87±69.94)比(44.23±27.52)ng/L,P=0.000]。外周血IL-8水平在COPD患者高于健康不吸烟者2倍以上[(103.98±82.39)比(44.23±27.52)ng/L,P=0.000]。外周血IL-10水平在健康吸烟者低于健康不吸烟者10%左右[(10.04±0.54)比(11.78±0.45)ng/L,P<0.05]。外周血IL-10水平在COPD患者低于健康不吸烟者10%左右[(10.12±0.56)比(11.78±0.45)ng/L,P<0.05]。外周血清IL-8水平与吸烟量呈线性正相关。IL-10水平与吸烟量零相关。吸烟量超过35包-年的COPD患者与吸烟量少于35包-年者相比,血清IL-8水平前者较后者较升高约3倍以上。结论:至少在轻中度COPD患者的全身炎症与吸烟密切相关。吸烟引起支气管-肺的炎症反应同时,可能通过抑制外周血单个核细胞中的HDAC活性造成全身炎症反应。

Objective： To explore the relation between the histone deacetylase （HDAC） activity in pe- ripheral blood mononuelear cells （PBMCs） and the systemic inflammation of COPD. Methods. The levels of serum interleukin-8 （IL-8） and interleukin 10 （IL-10） were determined by enzyme linked immunosorbent assay （ELISA） kit. The blood samples were isolated by density centrifugation within four hours. Histones were extracted from nuclei overnight by using HCl and H2 SO4 at 4℃ and using a method modified from that as described by other researchers. HDAC activity of nuclear extracts in PBMC was measured with an HDAC Activity/Inhibitor Screening Assay Kit. Results： HDAC activity in PBMC from COPD was negatively correlated with serums IL-8 level （r=-0. 693, P=0. 000 for patients with COPD, and r=-0. 598, P=0. 000 for all subjects）. The levels of serum interleukin-8 in healthy smokers wae higher than in healthy nonsmokers, as （106.87±69.94） vs （44.23±27.52） ng/L, P=0. 000, and in COPD patients too, that was （103.98±82.39）ng/L, P=0. 000. The level of serum IL-10 in healthy smoker was lower by 10G than which in healthy nonsmokers, as （10.04±0.54） vs （11.78±0.45） ng/L, P〈0.05, and in COPD patients too, that （10. 12±0. 56）ng/L, P〈0. 05. The positive correlation was found between smoking quality and serums IL-8. In COPD patients who smoke more than 35 pack years compared with those smoke less than 35 pack-years, the levels of IL-8 in former were three times higher than the later. Conclusion： It suggested that smoking contribute to the decreased HDAC activity in PBMC from COPD at least in mild and moderate stage. Smoking causes the local lung inflammation, at same time it also causes systemic inflammation through inhibiting the HDAC activity in PBMC from COPD.