LncRNA MSTRG.13,871/miR155-5p/Grip1 network involved in the post-cardiac arrest brain injury

Abstract

Post-cardiac arrest brain (PCABI) is a severe medical condition characterized by a significant risk of neurological impairment and death. Nevertheless, the specific process and essential molecules responsible for its development are not fully understood. Profiling based on competing endogenous RNAs (ceRNA) has been implicated in the onset and progression of neurological disorders, yet its role in PCABI remains unclear. In this study, we performed RNA transcriptome sequencing analysis to identify differentially expressed genes in the rat model for cardiac arrest and cardiopulmonary resuscitation (CA/CPR). A hub ceRNA regulatory network was constructed using miRWalk 2.0 and Cytohubba plug-in in Cytoscape. Subsequently, real-time quantitative reverse transcription-polymerase chain reaction and dual-luciferase activity assays validated MSTRG.13,871, miR-155-5p, and Grip1 as differentially expressed in CA/CPR group, with MSTRG.13,871 capable of targeting both miR-155-5p and Grip1. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed the ceRNA network enrichment in immunoregulation mechanisms such as mitochondrion, apoptotic process, and negative regulation cell death. Our research highlights the mechanism of PCABI by revealing a critical regulatory axis involving MSTRG.13,871-miR-155-5p-Grip1 in the hippocampus CA1 region after CA/CPR in rats, proposing a feasible controlled mechanism, which may serve as a theoretical basis for designing innovative therapies.

Keywords: Hippocampus; LncRNA; MiR-155-5p; Post-cardiac arrest brain injury; cardiopulmonary resuscitation.

Fig. 1

Animal preparation and experimental procedure.

Fig. 2

Screening of the DERNAs in the hippocampus tissues of CA/CPR group. (A-C) Volcano plot of DE lncRNAs (A), DE miRNAs (B), and DE mRNAs (C) (red, up-regulated; blue, down-regulated). (D-F) Cluster heat map of DE lncRNAs (D), DE miRNAs (E), and DE mRNAs (F) (red, up-regulated; blue, down-regulated).

Fig. 3

(A) Construction of the lncRNA-miRNA-mRNA regulatory network based on DE-mRNAs, DE-lncRNAs and DE-miRNAs. Red and blue represent up-regulated and down-regulated RNAs, respectively. Diamonds, triangles, and ellipses represent miRNAs, mRNAs, and lncRNAs, respectively. (B) Construction of the hub ceRNA regulatory network, including lncRNA MSTRG.13,871, 9 miRNAs and 15 mRNAs. The blue diamonds around lncRNA MSTRG.13,871 represent miRNA; the yellow triangles represent mRNA. (C) Classification of top 10 significant GO terms for hub ceRNA network. The results of enrichment cover three sets: biological process (BP), cellular component (CC), and molecular function (MF). (D) Significant KEGG pathways for hub ceRNA network.

Fig. 4

Validation of ceRNA regulatory network. (A) The relative expression level of lncRNA, miRNAs, and mRNAs involved in the ceRNA network using qRT-PCR. (B) Target relationship prediction between miR-155-5p and MSTRG.13,871 as well as Grip1; (C, D) Validation of the target relationships between miR-155-5p and MSTRG.13,871 as well as Grip1 using dual-luciferase assay. *p < 0.05, **P < 0.01, ****P < 0.0001.