CAJ | 학술논문

目的观察机械牵张力对人肺动脉内皮细胞(HPAEC)中细胞因子及黏附分子表达的影响,为探讨呼吸机相关性肺损伤(VILI)提供理论依据.方法采用机械牵张装置Flexcell FX-5000T对HPAEC细胞以0.5 Hz频率施加10％或20％牵张应力牵张3、6、12和24 h.应用实时荧光定量聚合酶链反应(qRT-PCR)、酶联免疫吸附试验(ELISA)以及蛋白质免疫印迹试验(Western blotting)检测机械牵张前后白细胞介素(IL-6、IL-8)、单核细胞趋化蛋白-1(MCP-1)等细胞因子及细胞间黏附分子-1(ICAM-1)的基因和蛋白表达.结果随牵张应力的增加,IL-8、MCP-1、ICAM-1的mRNA和蛋白表达随牵张时间的延长逐渐增加.与对照组(作为1)相比,20％牵张组牵张24 hHPAEC细胞的IL-8 mRNA表达上调至1.58±0.10,MCP-1mRNA表达上调至2.85±0.52,ICAM-1mRNA表达上调至1.90±0.14(均P＜0.05).与对照组相比,20％牵张组牵张24 h HPAEC细胞的IL-8、MCP-1蛋白表达(均ng/L)显著增加(IL-8∶3401.08±439.60比1422.60±66.98,MCP-1∶1117.64±237.54比307.88±80.84,均P＜0.05)；ICAM-1蛋白表达上调至2.15±0.40(P＜0.05)；而IL-6 mRNA和蛋白水平较对照组无明显变化.结论机械牵张可上调HPAEC细胞IL-8、MCP-1和ICAM-1的表达,这可能是机械通气致肺损伤的发病机制之一.
목적 관찰궤계견장력대인폐동맥내피세포(HPAEC)중세포인자급점부분자표체적영향,위탐토호흡궤상관성폐손상(VILI)제공이론의거.방법 채용궤계견장장치Flexcell FX-5000T대HPAEC세포이0.5 Hz빈솔시가10％혹20％견장응력견장3、6、12화24 h.응용실시형광정량취합매련반응(qRT-PCR)、매련면역흡부시험(ELISA)이급단백질면역인적시험(Western blotting)검측궤계견장전후백세포개소(IL-6、IL-8)、단핵세포추화단백-1(MCP-1)등세포인자급세포간점부분자-1(ICAM-1)적기인화단백표체.결과 수견장응력적증가,IL-8、MCP-1、ICAM-1적mRNA화단백표체수견장시간적연장축점증가.여대조조(작위1)상비,20％견장조견장24 hHPAEC세포적IL-8 mRNA표체상조지1.58±0.10,MCP-1mRNA표체상조지2.85±0.52,ICAM-1mRNA표체상조지1.90±0.14(균P＜0.05).여대조조상비,20％견장조견장24 h HPAEC세포적IL-8、MCP-1단백표체(균ng/L)현저증가(IL-8∶3401.08±439.60비1422.60±66.98,MCP-1∶1117.64±237.54비307.88±80.84,균P＜0.05)；ICAM-1단백표체상조지2.15±0.40(P＜0.05)；이IL-6 mRNA화단백수평교대조조무명현변화.결론 궤계견장가상조HPAEC세포IL-8、MCP-1화ICAM-1적표체,저가능시궤계통기치폐손상적발병궤제지일.
Objective To observe the effects of cyclic stretch on expression of cytokines and adhesion molecules in human pulmonary artery endothelial cells (HPAECs),herein to provide a theoretical basis to ventilator-induced lung injury (VILI).Methods HPAECs were subjected to cyclic stretch by the Flexcell FX-5000T system at 0.5 Hz of 10％ or 20％ elongation for 3,6,12,24 hours respectively.The mRNA and protein expression of interleukin (IL-6,IL-8),monocyte chemotactic protein-1 (MCP-1) and intercelluar adhesion molecule-1 (ICAM-1) was determined by fluorescent quantitation reverse transcription-polymerase chain reaction (qRT-PCR),enzyme linked immunosorbent assay (ELISA) or Western blotting.Results Increasing the stretch force,the mRNA and protein expression of IL-8,MCP-1,ICAM-1 were up regulated with increasing stretch time.Compared with the control (set 1),after 20％ cyclic stretch for 24 hours,IL-8 mRNA expression was up regulated to 1.58 ± 0.10,MCP-1 mRNA expression was up regulated to 2.85 ±0.52,and ICAM-1 mRNA expression was up regulated to 1.90 ±0.14 (all P＜0.05).Compared with control group,after 20％ cyclic stretch for 24 hours,the protein expression of IL-8 and MCP-1 (both ng/L) in HPAEC was significantly increased (IL-8 ∶ 3401.08 ±∶ 439.60 vs.1422.60 ± 66.98,MCP-1 ∶ 1117.64 ± 237.54vs.307.88 ± 80.84,both P＜0.05),ICAM-1 protein expression was up regulated to 2.15 ± 0.40 (P＜0.05),while the expression of IL-6 mRNA and protein had no statistic difference compared with control group.Conclusion Cyclic stretch enhanced the expression of IL-8,MCP-1 and ICAM-1 in an intensity-dependent fashion,so it may be involved in the pathogenesis of lung injury induced by mechanical ventilation.