目的:應(yīng)用全轉(zhuǎn)錄組芯片研究缺氧/復(fù)氧誘導(dǎo)下人臍靜脈內(nèi)皮細(xì)胞（HUVEC）的轉(zhuǎn)錄組輪廓。創(chuàng)新點:血管內(nèi)皮細(xì)胞（VEC）缺氧/復(fù)氧損傷被視定為許多生理和病理過程中導(dǎo)致器官功能障礙的重要驅(qū)動因素。然而,其詳細(xì)病理生理機制和基因表達(dá)譜信息尚未闡明。本研究首次應(yīng)用全轉(zhuǎn)錄組芯片技術(shù)研究VEC缺氧/復(fù)氧誘導(dǎo)下的轉(zhuǎn)錄組輪廓。方法:采用缺氧孵育3h后復(fù)氧1h的HUVEC為缺氧/復(fù)氧組,同時常氧孵育的HUVEC為常氧對照組。應(yīng)用含58 339條探針的全轉(zhuǎn)錄組芯片檢測每組三個樣本。對差異表達(dá)基因進(jìn)行生信分析和功能驗證。結(jié)論:本研究發(fā)現(xiàn)372個有意義的差異表達(dá)基因探針。相關(guān)基因涵蓋多種途徑和功能,例如氧自由基的產(chǎn)生、鈣超載、炎癥、糖脂代謝、內(nèi)皮細(xì)胞增殖、分化、細(xì)胞骨架及通透性調(diào)節(jié)、細(xì)胞裂解、凋亡和血管生成。另外,實驗進(jìn)一步表明,差異表達(dá)基因pleckstrin同源樣域家族A成員1（PHLDA1）的m RNA和蛋白質(zhì)表達(dá)結(jié)果與微陣列結(jié)果一致。STRING分析發(fā)現(xiàn),PHLDA1可能與差異表達(dá)基因SLC38A3、SLC5A5、Lnc-SLC36A4-1和Lnc-PLEKHJ1-1具有物理性和/或功能性相互作用,這有... 

【文章來源】：Journal of Zhejiang University-Science B（Biomedicine & Biotechnology）. 2020,21(04)SCICSCD

【文章頁數(shù)】：14 頁

【文章目錄】：
1 Introduction
2 Materials and methods
    2.1 Cell lines and cell culture
    2.2 Construction of transcriptome profile
    2.3 Analysis of microarray raw data
    2.4 Real-time PCR
    2.5 Western blot assay
    2.6 Protein interaction analysis
    2.7 Statistical analysis
3 Results
    3.1 Reliability and repeatability analyses of microarray-based hypoxia/re-oxygenation-induced HUVEC transcriptome profiling
    3.2 Hypoxia/re-oxygenation-related differentially expressed gene probes
    3.3 Pathways and functions mapping with hypoxia/re-oxygenation-sensitive gene probes
    3.4 Expression verification of differentially expressed gene PHLDA1 and exploration of related function based on HUVEC hypoxia/re-oxygenation transcriptome profiling
4 Discussion
5 Conclusions
Contributors
Compliance with ethics guidelines
List of electronic supplementary materials


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