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氧化应激抑制支气管上皮细胞CFTR表达的信号机制

发表时间:2010-12-15  浏览量:1448  下载量:564
全部作者: 屈飞,秦晓群,向阳,谭蓉,刘惠君
作者单位: 中南大学湘雅医学院生理学系;江西中医学院药学院
摘 要: 目的:研究氧化应激人支气管上皮细胞株(bronchial epithelial cells,BECs)后囊性纤维化跨膜转导调节因子(cystic fibrosis transmembrane conductance regulator,CFTR)表达调控的信号机制。方法:臭氧应激BECs后,实施聚合酶链反应(real-time polymerase chain reaction, real-time PCR)、免疫荧光和Western blot检测CFTR的表达。结果:Real-time PCR结果发现,臭氧应激后,AG490预处理能增加CFTR mRNA的表达,免疫荧光和Western blot结果显示,AG490预处理能增加CFTR蛋白的表达。Western blot结果显示,在未受臭氧处理的BECs,转录活化子1(signal transducer and activator of transcription 1,STAT1)蛋白的磷酸化水平很小,在臭氧处理后,STAT1的磷酸化水平随时间依赖性增加。应用小分子抑制剂预处理BECs,免疫荧光和Western blot结果显示,氨基胍(amino guanidine,AG)、锌卟啉-9(zinc protoporphyrin IX,ZnPP-9)、N-乙酰-L-半胱氨酸(N-acetylcysteine,NAC)均降低了臭氧诱导的STAT1磷酸化。Real-time PCR和Western blot结果显示,AG,NAC预处理能增加臭氧应激后CFTR的表达,而ZnPP-9则影响不明显。结论:NO、活性氧(reactive oxygen species,ROS)可通过STAT1通路的介导直接或间接影响CFTR的表达。
关 键 词: 生理学;囊性纤维化跨膜转导调节因子;臭氧应激;信号分子
Title: Mechanism of down-regulated CFTR expression by ozone stress on human bronchial epithelial cells
Author: QU Fei, QIN Xiaoqun1, XIANG Yang1, TAN Yurong1, LIU Huijun
Organization: Department of Physiology, Xiangya Medical School, Central South University; Department of Pharmacology, Jiangxi University of Traditional Chinese Medicine
Abstract: Objective: To investigate the signal mechanism of cystic fibrosis transmembrane conductance regulator (CFTR) expression by ozone stress on bronchial epithelial cells (BECs). Methods: Real-time polymerase chain reaction (PCR), immunofluorescence and western blot were detected the expression of CFTR after ozone stress. Results: Real-time PCR showed that pretreatment with AG490 attenuated the down-regulated CFTR mRNA expression induced by ozone stress. Immunofluorescence and western blot revealed that pretreatment with AG490 raised the CFTR protein expression. And it was also observed that ozone stress accelerated the phosphorylation of signal transducer and activator of transcription 1 (STAT1) in BECs. Furthermore, pretreatment with inhibiters, reactive N-acetylcysteine (NAC), amino guanidine (AG) and zinc protoporphyrin IX (ZnPP-9) could cut down the phosphorylation of STAT1. NAC and AG could down-regulate the expression of CFTR after ozone stress by immunofluorescence and western blot, but not ZnPP-9. Conclusion: NO and reactive oxygen species (ROS) can effect the expression of CFTR through STAT1 signal.
Key words: physiology; cystic fibrosis transmembrane conductance regulator; ozone stress; signaling molecule
发表期数: 2010年12月第23期
引用格式: 屈飞,秦晓群,向阳,等. 氧化应激抑制支气管上皮细胞CFTR表达的信号机制[J]. 中国科技论文在线精品论文,2010,3(23):2376-2380.
 
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