慢性缺氧改变外周血转录组模式

doi:10.24920/003598

摘要/Abstract

摘要：

目的多种生理和病理状态都会伴随着慢性缺氧,例如紫绀型先天性心脏病（CCHD）。这种慢性缺氧可能会干扰基因的转录过程。然而,在缺氧条件下外周血转录组模式的改变尚无报道。故本工作旨在探讨慢性缺氧条件下,外周血转录组模式的变化。
方法本研究使用慢性缺氧大鼠模型模拟CCHD患者的低氧状态。两组Sprague-Dawley大鼠（每组n=6）分别暴露于低氧（10%O₂）或常氧（21%O₂）条件下饲养3周。每周测量大鼠体重。缺氧处理结束后,采集两组大鼠外周血并提取总RNA进行RNA-Seq。经过质量评估后,通过Illumina Hiseq平台对文库进行测序。筛选差异表达基因（DEG）,筛选条件为FDR（false discovery rate）<0.05且FC（fold change）>2。进行DEG的功能注释和聚类分析,并选取出padj（adjusted P-value）<0.05的条目。
结果低氧组大鼠体重较对照组明显降低（P<0.01）。RNA-Seq结果表明：两组的转录组模式有显著差异。共鉴定出了872个差异表达的基因。在缺氧组中,共有803个基因下调,只有69个基因上调。对872个基因的功能富集分析表明：它们涉及了多个生物学过程,例如含卟啉的化合物代谢过程,血红蛋白复合物和氧转运蛋白活性等。
结论我们的研究表明慢性缺氧大鼠模型外周血的转录组模式发生了显著改变。为进一步了解CCHD患者的生理和病理变化提供了理论基础和研究方向。

关键词: 慢性缺氧, RNA-Seq, 外周血, 紫绀型先天性心脏病

Abstract:

Objective Many physiological and pathological conditions, including cyanotic congenital heart diseases (CCHD), are accompanied by chronic hypoxia, which might interfere with the transcription process. However, the transcriptome profile in peripheral blood under hypoxia is still unidentified. The present work aimed to explore the transcriptional profile alteration of peripheral blood in chronic hypoxia.
Methods The present study used a chronic hypoxia rat model to simulate the hypoxic state of CCHD patients. Two groups of Sprague-Dawley rats (n=6 per group) were either exposed to hypoxia (10% O₂) or normoxia (21% O₂) for 3 weeks. Body weight was measured weekly. Peripheral blood was collected and total RNA was extracted for RNA-Seq at the end of the hypoxia treatment. After quality assessment, the library was sequenced by the Illumina Hiseq platform. The differentially expressed genes were screened (false discovery rate<0.05 and fold change>2). The functional annotation analysis and cluster analysis of differentially expressed genes were performed based on the adjusted P-value (padj<0.05).
Results Compared with the control group, the body weight of the rats in the hypoxia group was significantly lowered (P<0.01). RNA-Seq results showed that the transcriptome patterns of the two groups had significant differences. In total, 872 genes were identified as differentially expressed. Among all, 803 genes were down-regulated, while only 69 genes were up-regulated in the hypoxia group. The functional enrichment analysis of the 872 genes showed that multiple biological processes involved, such as porphyrin-containing compound metabolic process, hemoglobin complex and oxygen transporter activity.
Conclusions Our study demonstrated the transcriptional profile alteration in peripheral blood of chronic hypoxia rat model. This study provided basic data and directions to further understand the physiological and pathological changes in patients with CCHD.

Key words: chronic hypoxia, RNA-Seq, peripheral blood, cyanotic congenital heart diseases

Wang Tingting, Xing Junyue, Zhang Lijing, Zhang Hao. Transcriptional Profile Alteration of Peripheral Blood in Chronic Hypoxia[J].Chinese Medical Sciences Journal, 2020, 35(1): 54-64.

图/表 7

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Sample ID	Exon	Intron	Intergenic	Sample ID	Exon	Intron	Intergenic
C1	99.61	0.18	0.20	H1	99.26	0.40	0.34
C2	99.22	0.44	0.34	H2	99.26	0.38	0.36
C3	99.48	0.27	0.25	H3	99.26	0.40	0.34
C4	99.45	0.28	0.27	H4	99.34	0.33	0.32
C5	99.07	0.54	0.39	H5	99.06	0.53	0.41
C6	99.48	0.27	0.25	H6	99.03	0.51	0.46

Genes	Degree	Log₂FC	Genes	Degree	Log₂FC
Ubb	56	1.54	Kctd7	22	1.74
Cdc20	37	1.84	Asb1	22	1.51
Anapc2	31	1.25	Trim21	22	1.44
Fzr1	30	1.36	Rnf25	22	1.15
Dctn2	26	1.37	Asb6	22	1.18
Dctn3	25	1.11	Ccnf	22	1.61
Ube2l6	25	1.94	Ube2e2	22	1.12
Aurka	23	1.48	Lmo7	22	1.07
Ube2o	23	1.20	Herc6	22	1.64
Nedd4l	23	1.17	Rnf123	22	1.17
Asb11	22	1.66	Traip	22	2.11
Fbxo7	22	1.75	Uba52	22	1.05
Rnf114	22	2.17	Pigz	20	2.35
Klhl25	22	1.88	Polr2e	19	1.85
Klhl42	22	1.21	Tubb4b	19	1.47

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