被动致敏对人气道平滑肌细胞Kv的影响及其PKC信号调控机制  

Changes of Kv in Passively Sensitized Human Airway Smooth Muscle Cells and Its Regulating Mechanism through PKC Signaling Pathway

作　　者：程东军[1] 徐永健[1] 刘先胜[1] 赵丽敏[1] 熊盛道[1] 张珍祥[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院呼吸内科,武汉430030

出　　处：《华中科技大学学报（医学版）》2006年第2期150-154,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30270583)

摘　　要：目的研究哮喘患者血清被动致敏人气道平滑肌细胞(HASMCs)后电压依赖延迟整流钾通道(Kv)的变化,探讨其蛋白激酶C(PKC)信号调控机制。方法采用全细胞膜片钳、逆转录聚合酶链反应(RT-PCR)、免疫细胞化学和Western blot技术,观察人哮喘血清被动致敏后的HASMCs Kv活性和PKC-α及Kv1.5亚型蛋白质表达的变化,以及PKC对HASMCs的Kv活性和Kv1.5亚型mRNA及蛋白质表达的影响。结果被动致敏后的HASMCs的PKC-αmRNA和蛋白质表达升高,Kv活性及其Kv1.5亚型mRNA和蛋白质的表达均下降;PKC激活剂豆蔻酰佛波醇乙酯(Phorbol 12-myristate 13-acetate,PMA)显著抑制HASMCs的Kv电流活性,降低Kv1.5亚型的表达,该效应可被PKC阻断剂Ro31-8220显著逆转。结论被动致敏的HASMCs的Kv活性降低,可能与PKC活化后的直接作用及进而下调Kv表达有关。Objective To investigate the changes of voltage dependent delayed rectifier potassium channel （Kv） on human airway smooth muscle cells （HASMCs） passively sensitized with asthmatics serum and its regulating mechanism by protein kinase C （PKC） signaling pathway. Methods Kv activities of HASMCs passively sensitized with asthmatics serum were investigated with whole-cell patch clamp. The mRNA and protein expression of PKC-α and Kv1. 5 in passively sensitized HASMCs was detected by using techniques of immunocytochemistry, reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting. The effect of PKC signaling pathway on the activity and expression of Kv in passively sensitized HASMCs were also investigated. Results After HASMCs were sensitized with asthmatics serum, the mRNA and protein expression of PKC-α was increased, and Kv activities and Kv1. 5 expression were decreased. Phorbol 12-myristate 13-acetate （PMA）, a PKC activator, caused a significant inhibition of Kv currents and the expression of Kv1. 5 in cultured HASMCs, and the inhibition was partly abolished by Ro31 8220, a PKC inhibitor. Conclusion The function of Kv in passively sensitized HASMCs is decreased, which may directly result from PKC activation and subsequent down regulation of the Kv1. 5 expression.

关 键 词：电压依赖延迟整流钾通道 人气道平滑肌 蛋白激酶C 哮喘 

分 类 号：R562.25[医药卫生—呼吸系统]