SARS-CoV的S蛋白诱导呼吸道上皮细胞合成释放IP-10的信号分子机制研究  

SARS-CoV S protein induces chemokine IP-10 expression in airway epithelial cells

作　　者：要国华[1] 杨新艳[1] 姜勇[2] 徐军[1] 

机构地区：[1]广州医学院附属第一医院广州呼吸疾病研究所,广东广州510120 [2]南方医科大学广东省功能蛋白质组学重点实验室,广东广州510515

出　　处：《中国病理生理杂志》2008年第3期423-426,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30340029);广东省教育厅非典专项经费;广东省非典攻关病原学专项基金资助项目(No.303001010)

摘　　要：目的:研究SARS冠状病毒S蛋白诱导呼吸道上皮细胞合成释放IP-10(interferon-gamma induc-ible protein 10)的信号分子机制。方法:通过基因芯片检测SARS冠状病毒的S蛋白作用于人支气管上皮细胞16HBE后信号通路基因表达谱的变化;采纳RT-PCR、EMSA、Western blotting等方法进一步分析JAK-STAT通路中信号分子的磷酸化、IRF-1和IP-10基因表达的变化及其相应信号分子抑制剂对表达水平的影响。结果:S蛋白作用于人支气管上皮细胞16HBE诱导了JAK-STAT信号通路涉及的重要转录因子基因IRF-1的表达,该信号通路的转录因子STAT1在刺激后15 min发生磷酸化,2 h即可检出IP-10基因的表达,IP-10的表达可以完全被STAT1、JAK2抑制剂阻断。EMSA显示:支气管上皮细胞在S蛋白的作用下,其核蛋白能够特异性与ISRE和GASDNA基序相结合,而不能与NF-κB的DNA基序相结合。结论:SARS-CoV的S蛋白通过激活JAK-STAT信号转导通路诱导IP-10在宿主细胞的生成。提示病毒诱导的JAK-STAT信号通路激活在病毒感染相关的急性肺损伤发生中具有重要地位。AIM ： To investigate the molecular mechanism by which the SARS - CoV S protein induces chemo- kine IP - 10 in airway epithelial cells. METHODS： cDNA microarrays were used to screen the gene expression profiles of human bronchial epithelial cells （16HBEs） stimulated by SARS- CoV S protein. In addition, RT- PCR, EMSA, and Western blotting were performed to analyze the phosphorylation of JAK/STAT signal pathway. The changes of IRF - 1 and IP - 10 gene expression and the influence by the corresponding inhibitors were analyzed. RESULTS ： IRF - 1, a key transcription factor of the JAK/STAT signal pathway, was activated in human bronchial epithelial cells after stimulation by the S protein of SARS - CoV. The IP - 10 gene expression was detected 2 h following the phosphorylation of STAT1 after 15 min, which was blocked by STATlor JAK2 inhibitors. Electrophoretic mobility shift assay （EMSA） demonstrated that the nuclear proteins bound to ISRE and GAS but not NF - KB DNA motif. CONCLUSION： The SARS - CoV S protein induces IP - 10 gene expression in human bronchial epithelial cells through activation of the JAK/STAT signal pathway, suggesting that the JAK/STAT signal pathway activated by virus plays key roles in virus infection related acute lung injury.

关 键 词：严重急性呼吸综合征 冠状病毒属 S蛋白 气道上皮细胞 γ干扰素诱导蛋白10 

分 类 号：R363[医药卫生—病理学]