CAJ | 학술논문

目的：探讨慢性阻塞性肺疾病（COPD）患者外周血中的早期内皮祖细胞（early-EPCs）是否表达β2-肾上腺素受体（β2 AR）及其对细胞迁移的影响。方法：晨起从COPD患者与对照组抽取20 mL静脉血，用Ficoll密度梯度＋磁珠分离法提取CD34＋细胞。通过Western blotting、RT-PCR及流式细胞术测定early-EPCs的β2 AR表达。体外培养early-EPCs，应用β2 AR的激动剂、抑制剂及下调β2 AR的表达量，通过Transwell法测定early-EPCs的迁移能力。结果：来自COPD患者的early-EPCs可见β2 AR mRNA及蛋白的表达强于对照组细胞（ P＜0.01）。两组细胞均表达β2 AR，但COPD组明显高于对照组，流式细胞术分析两组的β2 AR阳性细胞分别是82．9％和55．7％。外源性β2 AR抑制剂、β2 AR抗体及下调β2 AR的表达均可改善COPD患者early-EPCs 的迁移能力。结论：β2 AR在COPD患者及对照组外周血中的early-EPCs均有表达。降低β2 AR的表达可影响COPD患者early-EPCs的迁移功能。
목적：탐토만성조새성폐질병（COPD）환자외주혈중적조기내피조세포（early-EPCs）시부표체β2-신상선소수체（β2 AR）급기대세포천이적영향。방법：신기종COPD환자여대조조추취20 mL정맥혈，용Ficoll밀도제도＋자주분리법제취CD34＋세포。통과Western blotting、RT-PCR급류식세포술측정early-EPCs적β2 AR표체。체외배양early-EPCs，응용β2 AR적격동제、억제제급하조β2 AR적표체량，통과Transwell법측정early-EPCs적천이능력。결과：래자COPD환자적early-EPCs가견β2 AR mRNA급단백적표체강우대조조세포（ P＜0.01）。량조세포균표체β2 AR，단COPD조명현고우대조조，류식세포술분석량조적β2 AR양성세포분별시82．9％화55．7％。외원성β2 AR억제제、β2 AR항체급하조β2 AR적표체균가개선COPD환자early-EPCs 적천이능력。결론：β2 AR재COPD환자급대조조외주혈중적early-EPCs균유표체。강저β2 AR적표체가영향COPD환자early-EPCs적천이공능。
AIM:To investigate whether early endothelial progenitor cells (early-EPCs) expressβ2-adrenergic receptor (β2 AR) in the chronic obstructive pulmonary disease ( COPD) patients and the effect of β2 AR expression on the migration of early-EPCs.METHODS:Venous blood samples (20 mL) were obtained from antecubital vein of COPD pa-tients or healthy controls .Peripheral blood mononuclear cells were isolated by standard Ficoll gradient centrifugation , and purified by CD34 positive selection cocktail .The mRNA expression of β2 AR in the early-EPCs was detected by RT-PCR. The protein levels of β2 AR were assessed by Western blotting and flow cytometry .Chemotaxis was studied by Transwell as-say.Cultured early-EPCs were treated with ICI118551, norpinephrine (NE) or monoclonal antibody of β2AR (mAb-β2 AR) for 24 h.The number of migratory cells was counted under a light microscope .RESULTS:The level of β2 AR ex-pression in the COPD patients was higher than that in the controls .The number of migratory early-EPCs to stromal cell-de-rived factor 1αwas significantly improved by ICI 118551 compared with other COPD groups .When early-EPCs from the COPD patients or the controls were treated with different concentrations of mAb-β2 AR for 24 h, the number of migratory early-EPCs from the COPD patients and the controls treated with NE at concentration of 100 nmol/L was significantly re-duced.However, a marked decrease in the number of migratory early-EPCs from the COPD patients treated with NE was observed compared with control group .Before treated with ICI118551 or NE for 24 h, the early-EPCs were co-incubated with mAb-β2 AR for 40 min, and the number of migratory early-EPCs was not significantly different between COPD group and control group .Genetic down-regulation of β2 AR promoted the migration of early-EPCs in COPD group .CONCLU-SION:The level of β2 AR expression in the COPD patients is increased compared with the controls .The down-regulation ofβ2 AR improves the migration of early-EPCs.