脂质体转染Kv1.5反义寡核苷酸(AsOND)对人气道平滑肌Kv活性的影响

Effect of liposome-mediated transfection of Kv1.5 antisense oligonucleotides on activity of Kv in airway smooth muscle cells

目的:研究人气道平滑肌细胞(HASMCs)转染Kv1.5反义寡核苷酸(AsOND)后,电压依赖延迟整流钾通道(Kv)的活性变化,探讨其基因亚型Kv1.5在调节Kv活性中的作用。方法:采用脂质体转染、逆转录聚合酶链反应(RT-PCR)、Western blotting和全细胞膜片钳技术,观察转染Kv1.5 AsOND后,HASMCs Kv1.5 mRNA及蛋白表达的变化,以及转染对HASMCs Kv活性的影响。结果:脂质体转染Kv1.5 AsOND后,HASM细胞Kv1.5mRNA和蛋白质的表达均下降;Kv电流值受到显著抑制;使细胞膜电位(Em)趋向于去极化方向。结论:转染Kv1.5 AsOND可导致HASMCs Kv功能降低,Kv1.5基因亚型在调节Kv活性中可能起重要作用。

AIM ： To investigate the activity change of voltage - dependent delayed rectifier potassium channel （Kv） on human airway smooth muscle cells （HASMCs） after transfection of Kv1. 5 antisense oligonucleotides （ AsOND）, and to discuss the regulating mechanism of Kv1. 5. METHODS： The mRNA and protein expressions of Kv1. 5 in lipesome - mediated Kv1. 5 AsOND transfected HASMCs were measured with techniques of reverse transcriptase - polymerase chain reaction （ RT - PCR） and Western blotting. Kv activities in transfected HASMCs were investigated with whole - cell patch clamp. RESULTS： After HASMC were transfected by liposome - mediated Kv1. 5 AsOND, the mRNA and protein expressions of Kv1. 5 were decreased, and Kv activity was inhibited, which made the cell membrane potential （Em） inclined to depolarization. CONCLUSION： Transfection of Kv1. 5 AsOND made the function of Kv in HASMCs decreased. Kv1. 5 may play a critical role in the regulation of Kv activity.