机械牵张对人肺动脉内皮细胞中细胞因子及细胞间黏附分子-1表达的影响

Effects of cyclic stretch on expression of cytokines and intercelluar adhesion molecule-1 in human pulmonary artery endothelial cell

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.

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.