基于微阵列技术的缺氧/复氧诱导下血管内皮细胞转录组分析（英文）

发布时间：2021-07-12 01:43

　　目的:应用全转录组芯片研究缺氧/复氧诱导下人脐静脉内皮细胞（HUVEC）的转录组轮廓。创新点:血管内皮细胞（VEC）缺氧/复氧损伤被视定为许多生理和病理过程中导致器官功能障碍的重要驱动因素。然而,其详细病理生理机制和基因表达谱信息尚未阐明。本研究首次应用全转录组芯片技术研究VEC缺氧/复氧诱导下的转录组轮廓。方法:采用缺氧孵育3h后复氧1h的HUVEC为缺氧/复氧组,同时常氧孵育的HUVEC为常氧对照组。应用含58 339条探针的全转录组芯片检测每组三个样本。对差异表达基因进行生信分析和功能验证。结论:本研究发现372个有意义的差异表达基因探针。相关基因涵盖多种途径和功能,例如氧自由基的产生、钙超载、炎症、糖脂代谢、内皮细胞增殖、分化、细胞骨架及通透性调节、细胞裂解、凋亡和血管生成。另外,实验进一步表明,差异表达基因pleckstrin同源样域家族A成员1（PHLDA1）的m RNA和蛋白质表达结果与微阵列结果一致。STRING分析发现,PHLDA1可能与差异表达基因SLC38A3、SLC5A5、Lnc-SLC36A4-1和Lnc-PLEKHJ1-1具有物理性和/或功能性相互作用,这有... 

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

【文章页数】：14 页

【文章目录】：
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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