Screening and validation of differentially expressed microRNAs and target genes in hypertensive mice induced by cytomegalovirus infection

巨细胞病毒感染致高血压小鼠差异表达microRNA及靶基因的筛选与验证

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作者:YunZhong Shi #, DongMei Xi #, XiaoNi Zhang, Zhen Huang, Na Tang, YongMin Liu, LaMei Wang, Yan Tang, Hua Zhong, Fang He

Conclusion

This novel regulatory axis could aid the development of new approaches for the clinical prevention and control of EH.

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.

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