应用生物芯片技术探讨疟原虫MIF的信号通路作用机制  

APPLICATION OF BIO-CHIP TECHNIQUE TO EXPLORE THE MECHANISM OF SIGNAL TRANSDUCTION PATHWAYS BY PLASMODIUM-DERIVED MIF

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作  者:钟翔[1] 曾山[1] 邵丁丁[1] 王恒[1] 

机构地区:[1]中国医学科学院基础医学研究所,北京协和医学院基础学院,北京100005

出  处:《寄生虫与医学昆虫学报》2009年第2期76-81,共6页Acta Parasitologica et Medica Entomologica Sinica

基  金:国家自然科学基金项目(No.30700761);973项目(No.2007CB513100)

摘  要:通过生物芯片的方法,寻找约氏疟原虫(Plasmodium yoelii)来源的巨噬细胞迁移抑制因子(Macrophage migration inhibitory factor,MIF)(PyMIF)和宿主小鼠来源的MIF分子(MmMIF)在调节宿主细胞信号通路方面的差异,从而研究这2种MIF分子在作用机制上的异同。将亲和纯化原核表达的带His标签的重组MIF蛋白,通过C8柱去除内毒素后,刺激体外培养的小鼠单核/巨噬细胞系,纯化高质量的RNA,然后通过信号发现者通路芯片进行分析。结果表明PyMIF可能参与了多个细胞存活和抗凋亡相关的通路并上调了一些炎症基因的高转录,扩大了炎症反应并参与了维持靶细胞活性信号通路的转导。Application of bio-chip technique to find the differences of signal pathway between Plasmodium yoelii derived macrophage migration inhibitory factor (PyMIF) and murine macrophage migration inhibitory factor (MmMIF), so as to deduce the different mechanism of the two MIFs. We first purified the recombinant MIFs with His tag and removed the endotoxin by using C8 reversed-phase column. Then the endotoxin-free protein was added to cultured mouse monocyte/macrophage cell line, from which we purified high-quality RNA, and then proceed to "Signal Transduction Pathway Finder" microarray analysis. The results showed that PyMIF may be involved in cell survival and anti-apoptosis pathways and up-regulating some inflammatory gene transcription, which suggested that PyMIF may expand the inflammatory response and participate in some signal transduction pathways to maintain the target cells survival.

关 键 词:巨噬细胞迁移抑制因子 疟原虫 基因芯片 信号通路 

分 类 号:Q503[生物学—生物化学] Q257

 

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