CAJ | 학술논문

背景嗜酸粒细胞(EOS)在哮喘气道炎症中起着很重作用,我们前期发Slingshot-1L(SSH-1L)在急性发作期哮喘病人外周血EOS表达明显增高,可能在EOS的激活、迁移中发挥重要作用.目的探讨SSH-1L在哮喘急性发作期病人外周血EOS的定位及治疗后的表达及功能变化.方法招募4例未经糖皮质激素治疗的急性发作期哮喘门诊病人,在治疗前及吸入糖皮质激素3个月后抽取外周静脉血各30ml进行EOS的分离纯化并计数,应用RT-PCR和WesternBlotting方法分别在基因及蛋白质水平检测哮喘外周血EOS SSH-1L的表达变化;应用免疫荧光观察其中1例病人SSH-1L在外周静脉血EOS中的定位.结果免疫荧光显示外周血EOS的SSH-1L定位在胞浆中,在贴近胞壁处丰富表达.急性发作期哮喘病人糖皮质激素治疗前后外周血EOS RT-PCR扩增SSH-1L基因的SSH-1L/β-Actin灰度值比:治疗前0.7403±0.1124,治疗后0.4101±0.0363,治疗前后有显著差异(P=-0.001);急性发作期哮喘病人吸入糖皮质激素治疗前后外周血EOS蛋白的SSH.1L/β-actin灰度值比分别为0.3410±0.1337和0.1543±0.0551,治疗后明显降低(P=0.039).结论 SSH-1L可能在急性发作期哮喘病人外周血EOS的激活、迁移上发挥作用,糖皮质激素可能通过抑制哮喘急性发作期EOS的SSH表达而抑制Eos的激活、迁移起到治疗作用.
배경기산립세포(EOS)재효천기도염증중기착흔중작용,아문전기발Slingshot-1L(SSH-1L)재급성발작기효천병인외주혈EOS표체명현증고,가능재EOS적격활、천이중발휘중요작용.목적 탐토SSH-1L재효천급성발작기병인외주혈EOS적정위급치료후적표체급공능변화.방법 초모4례미경당피질격소치료적급성발작기효천문진병인,재치료전급흡입당피질격소3개월후추취외주정맥혈각30ml진행EOS적분리순화병계수,응용RT-PCR화WesternBlotting방법분별재기인급단백질수평검측효천외주혈EOS SSH-1L적표체변화;응용면역형광관찰기중1례병인SSH-1L재외주정맥혈EOS중적정위.결과 면역형광현시외주혈EOS적SSH-1L정위재포장중,재첩근포벽처봉부표체.급성발작기효천병인당피질격소치료전후외주혈EOS RT-PCR확증SSH-1L기인적SSH-1L/β-Actin회도치비:치료전0.7403±0.1124,치료후0.4101±0.0363,치료전후유현저차이(P=-0.001);급성발작기효천병인흡입당피질격소치료전후외주혈EOS단백적SSH.1L/β-actin회도치비분별위0.3410±0.1337화0.1543±0.0551,치료후명현강저(P=0.039).결론 SSH-1L가능재급성발작기효천병인외주혈EOS적격활、천이상발휘작용,당피질격소가능통과억제효천급성발작기EOS적SSH표체이억제Eos적격활、천이기도치료작용.
Background Eosinophils play a pivotal role in asthmatic airway inflammation. We previously found a significantly high expression of Slingshot-IL (SSH-1L) in peripheral eosinophils in acute exacerbations of asthma. Objective To investigate the expression and localization patterns of SSH-1L in peripheral blood eosinophils of asthmatic patients and their changes after treatment with inhaled corticosteroids. Methods We recruited 4 outpatients with acute exacerbations of asthma who received no previous corticosteroid treatment and 1 healthy volunteer. From all the subjects 30 ml peripheral venous blood samples were collected before and after a 3-month treatment with inhaled fluticasone. The eosinophils were isolated, purified and counted, and the expressions of SSH-1L in the eosinophils were examined by RT-PCR and Western blotting. The localization of SSH-1L phosphatases in the peripheral eosinophils was detected by immunofluorescence assay in one patient. Results SSH-1L phosphatases distributed diffusely in the cytoplasm, especially dense near the membrane of the peripheral eosinophils. Glucocorticoids treatment resulted in a significant reduction in both the SSH-1L mRNA expression (0.7403 ± 0.1124 vs 0.4101 ± 0.0363, P=0.001) and SSH-1L protein expression (0.3410 ± 0.1337 vs 0.1543 ± 0.0551, P=0.039). Conclusions A high expression of SSH-1L in peripheral eosinophils in acute exacerbations of asthma may play a role in the activation and migration of eosinophils. The efficacy of inhaled corticosteroids in asthma control might be partly attributed to a down-regulated expression of SSH-1L.