Screening and validation of differentially expressed microRNAs and target genes in hypertensive mice induced by cytomegalovirus infection.
Screening and validation of differentially expressed microRNAs and target genes in hypertensive mice induced by cytomegalovirus infection.
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巨细胞病毒感染致高血压小鼠差异表达microRNA及靶基因的筛选及验证
DOI：
10.1042/bsr20202387
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发表时间：
2020-12-23
影响因子：
4
通讯作者：
He F
中科院分区：
文献类型：
--
作者：
Shi Y;Xi D;Zhang X;Huang Z;Tang N;Liu Y;Wang L;Tang Y;Zhong H;He F
摘要引言：多项研究表明巨细胞病毒（CMV）感染与原发性高血压（EH）之间存在关联。microRNAs（miRNAs）通过调控特定靶基因的表达在EH的发生发展中起着重要作用。然而，关于miRNA在CMV诱导的EH中的作用知之甚少。在本研究中，我们比较了正常和鼠巨细胞病毒（MCMV）感染的C57 BL/6小鼠样本的miRNA表达谱使用高通量测序分析。方法：取20只正常小鼠和20只MCMV感染小鼠的胸主动脉、心脏组织和外周血。我们在外周血样本中鉴定了差异表达的miRNAs，并利用生物信息学工具预测了它们的靶基因。然后，我们用定量逆转录聚合酶链反应（qRT-PCR）和双荧光素酶报告基因测定的靶基因进行实验验证。结果如下：结果发现118个差异表达的miRNAs，其中9个miRNAs被鉴定为MCMV感染性高血压的潜在调节因子。然后，我们使用qRT-PCR验证了两种候选miRNAs mmu-miR-1929- 3 p和mcmv-miR-m01-4- 5 p的表达。此外，双荧光素酶报告基因检测显示内皮素A受体（Ednra）mRNA的3′-非翻译区（UTR）含有mmu-miR-1929- 3 p的结合位点。总的来说，我们的数据表明MCMV感染可以升高C57 BL/6小鼠的血压并降低mmu-miR-1929- 3 p表达。mmu-miR-1929- 3 p靶向Ednra mRNA的3′-UTR。结论：这一新的调节轴可能有助于开发临床预防和控制高血压的新方法。
Abstract Introduction: Multiple studies have suggested an association between cytomegalovirus (CMV) infection and essential hypertension (EH). MicroRNAs (miRNAs) play a critical role in the development of EH by regulating the expression of specific target genes. However, little is known about the role of miRNAs in CMV-induced EH. In the present study, we compared the miRNA expression profiles of samples from normal and murine cytomegalovirus (MCMV)-infected C57BL/6 mice using high-throughput sequencing analysis. Methods: We collected the thoracic aorta, heart tissues, and peripheral blood from 20 normal mice and 20 MCMV-infected mice. We identified differentially expressed miRNAs in the peripheral blood samples and predicted their target genes using bioinformatics tools. We then experimentally validated them using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and the target genes with double luciferase reporter gene assay. Results: We found 118 differentially expressed miRNAs, among which 9 miRNAs were identified as potential MCMV infection-induced hypertension regulators. We then validated the expression of two candidate miRNAs, mmu-miR-1929-3p and mcmv-miR-m01-4-5p, using qRT-PCR. Furthermore, the dual-luciferase reporter gene assay revealed that the 3′-untranslated region (UTR) of endothelin A receptor (Ednra) messenger RNA (mRNA) contained a binding site for mmu-miR-1929-3p. Collectively, our data suggest that MCMV infection can raise the blood pressure and reduce mmu-miR-1929-3p expression in C57BL/6 mice. Moreover, we found that mmu-miR-1929-3p targets the 3′-UTR of the Ednra mRNA. Conclusion: This novel regulatory axis could aid the development of new approaches for the clinical prevention and control of EH.