反义核酸对气道平滑肌细胞内皮素转换酶-1基因表达的影响

Effect of Antisense Oligonecleotide on Endothelin Converting Enzyme Expression from Airway Smooth Muscle Cell

目的：探讨肿瘤坏死因子（TN－α）对人气道平滑肌细胞内皮素转换酶－1（ECE－1）基因表达的影响及体外反义内皮素转换酶－1寡聚核苷酸（ECE—1oDNS）是否可拮抗TNF－α诱导人气道平滑肌细胞ECE－1表达分泌．方法：高体培养人气过平滑肌细胞共导入由阳离子脂质体Lipofectin携载的不同浓度反义ECEoDNS，用RTPCR观察其对前炎症因子TNF-α诱导人气道平滑肌细胞ECE基因表达的影响．结果：TNF－α可刺激ECE-1mRNA表达．ECE－1mRNA由0．1318±0．0318升至0．2277±0．0150（t=4．724，P＜0．01）；导入反义ECEoDNS后，ECE－1mRNA表达受到抑制，抑制率分别为34．74％±11．6％及29．24％±4.96％．结论：反义ECE－1oDNS能拮抗TNF－α诱导人气道平滑肌细胞的ECE—1mRNA表达，为从基因治疗角度探讨控制气道局反应提供新线索．

Objective:To investigate the effects of endothelin converting enzyme (ECE) antisenseoligonuceotides (oDNS) on TNF α induced ECE expression from human airway smooth muscle cell(ASMC).Method:ASMC transfected with the antisense oligonucleodides of ECE vectored withlipofectin were established.TNF a induced ECE mRNA expression from ASMC were determined byRT-PCR. Result:ECE expression (ECE/β-actin: 0.2277±0.0150) in culture of ASMC incubated with100u／ml TNF α were significantly higher than those in control without TNF α incubation(0.1318±0.0318 t=4.724, P<0.01). However, after transfecting antisense ECEoDNS in those cells, ECEexpressions were significantly lower than those in control cells wthout antisense transfection.The inhibi-tion rate were 29.24%±4.96% and 34.74%±11.6% respectively. Conclusion: TNF α inducedECE mRNA expression from human ASMC are inhibited by antisense oligonucleotide of ECE.