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. 2019 May;19(5):4175-4184.
doi: 10.3892/mmr.2019.10073. Epub 2019 Mar 21.

Identification and functional analysis of microRNAs in rats following focal cerebral ischemia injury

Xianchun Duan  1 , Jianghua Gan  1 , Dai-Yin Peng  1 , Qiuyu Bao  2 , Ling Xiao  2 , Liangbing Wei  1 , Jian Wu  1
Affiliations

Identification and functional analysis of microRNAs in rats following focal cerebral ischemia injury

Xianchun Duan et al. Mol Med Rep. 2019 May.

Abstract

MicroRNA sequencing (miRNA‑seq) was performed in the present study to investigate miRNA expression profiles in infarcted brain areas following focal cerebral ischemia induced by middle cerebral artery occlusion in rats. In total, 20 miRNAs were identified to be upregulated and 17 to be downregulated in the infarct area. The expression levels of six differentially expressed miRNAs (DEmiRs), miR‑211‑5p, miR‑183‑5p, miR‑10b‑3p, miR‑182, miR‑217‑5p and miR‑96‑5p, were examined by reverse transcription‑​quantitative polymerase chain reaction. Subsequently, a miRNA‑mRNA network was constructed. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to investigate the functions of the mRNAs targeted by these DEmiRs. The present study aimed to investigate the association between miRNAs and cerebral ischemia to provide potential insight into the molecular mechanisms underlying ischemic stroke.

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Figures

Figure 1.
Figure 1.
MCAO-induced rat cerebral injuries, as detected by hematoxylin and eosin staining. (A) Section of the brain of a rat in the control group. (B) Section of the brain of a rat following MCAO. Magnification of the area corresponding to the injury region in the (C) control group and in the (D) MCAO group. MCAO, middle cerebral artery occlusion.
Figure 2.
Figure 2.
RNA-seq data corresponding to the DEmiRNAs between the MCAO model and control groups. (A) Hierarchical cluster of DEmiRNAs in the control vs. model groups. Green in the heat map represents downregulation and red represents upregulation. The average signals of the DEmiRNAs in each group were clustered using a Euclidean distance function. miRNAs exhibiting similar expression patterns are clustered together. n=3 in each group. (B) Volcano plots of transformed P-values (log10) against log2fold changes corresponding to transcripts per million reads in the two conditions examined. Blue and red dots represent differentially expressed miRNAs, whereas grey dots represent miRNAs that are not differentially expressed. RNA-seq, RNA sequencing; miRNA, microRNA; DEmiRNAs, differentially expressed microRNAs.
Figure 3.
Figure 3.
Levels of expression of six miRs assessed by reverse transcription-quantitative polymerase chain reaction. n=3 in each group. *P<0.05 vs. sham control. miR, microRNA.
Figure 4.
Figure 4.
GO terms enriched in genes targeted by the differentially expressed microRNAs. GO, Gene Ontology.
Figure 5.
Figure 5.
Top 30 most significant GO terms enriched in genes targeted by the differentially expressed microRNAs. GO, Gene Ontology.
Figure 6.
Figure 6.
KEGG terms enriched in genes targeted by the differentially expressed microRNAs. KEGG, Kyoto Encyclopedia of Genes and Genomes.
Figure 7.
Figure 7.
Top 30 KEGG pathways enriched in genes targeted by the differentially expressed microRNAs. KEGG, Kyoto Encyclopedia of Genes and Genomes.
Figure 8.
Figure 8.
miRNA-mRNA network constructed using six differentially expressed miRNAs that were confirmed by reverse transcription-quantitative polymerase chain reaction. Red indicates upregulated miRNAs and green indicates downregulated miRNAs. Pink elliptical nodes represent mRNAs.
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