Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury

Hongchang Yang; Jing Zhou; Junlei Wang; Luoning Zhang; Quzhi Liu; Jing Luo; Hongyan Jia; Li Liu; Qiang Zhou

doi:10.22074/cellj.2021.7275

Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury

Cell J. 2021 Sep;23(4):474-484. doi: 10.22074/cellj.2021.7275. Epub 2021 Aug 29.

Authors

Hongchang Yang^#¹, Jing Zhou^#², Junlei Wang³, Luoning Zhang¹, Quzhi Liu⁴, Jing Luo⁵, Hongyan Jia⁶, Li Liu⁷, Qiang Zhou⁸

Affiliations

¹ Physical Education Department, Hohai University, Nanjing, Jiangsu, China.
² Department of Clinical Medicine, Jiangsu Health Vocational College, Nanjing, Jiangsu, China.
³ Harbor, Channel and Coastal Engineering, Hohai University, Nanjing, Jiangsu, China.
⁴ Centre of Counseling and Psychological Services, Hohai University, Nanjing, Jiangsu, China.
⁵ Center for Kidney Disease, 2nd Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, China.
⁶ Port Channel and Coastal Engineering Department, Hohai University, Nanjing, Jiangsu, China.
⁷ The Department of Rehabilitation, Brain Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu, China. Email: liulicao1976@163.com.
⁸ Physical Education Department, Hohai University, Nanjing, Jiangsu, China. Email: 19870073@hhu.edu.cn.

^# Contributed equally.

Abstract

Objective: This study aimed to characterize the circulating exosomal microRNA (miRNA) profiles associated with acute soft tissue injury.

Materials and methods: In this experimental study, a total of 12 rats were randomly divided into control group and model group (n=6 for each group). The rats in the model group were used to establish an acute soft tissue injury following the mechanical injury of the leg. The exosomes from the peripheral blood of all the rats were isolated and then characterized by Nanosight NS300 particle size analyser (NTA), transmission electron microscopy (TEM) and western blot. Next, the exosomal miRNAs in the control and model groups were sequenced, and the differentially expressed miRNAs (DE-miRNAs) were identified using the DESeq algorithm. Functional analyses were performed using Gene Ontology (GO) terms and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway databases. Finally, quantitative reverse-transcription polymersa chain reaction (qRT-PCR) was used to verify the expression of the DE-miRNAs.

Results: TEM, NTA and western blot results showed that the exosomes were approximately 100 nm in size and exhibited cup-shaped morphology. A total of 628 miRNAs were obtained by sequencing. After that, 28 DE miRNAs (DEmiRNAs) were identified, including seven down-regulated miRNAs and 21 up-regulated miRNAs. These DE-miRNAs were linked to 7539 target genes with GO. Also, KEGG analyses demonstrated that these genes were enriched for phosphorylation, VEGF signaling pathway, and MAPK signaling pathway. Additionally, the consistency rate between the qRT-PCR and sequencing results was 83.33%, which showed a high relative reliability of the sequencing results.

Conclusion: These findings suggest that these 28 exosomal miRNAs may be involved in the regulation of acute soft tissue injury, by one of critical biological processes (BP), phosphorylation. The findings provide valuable clues by utilizing exosomes as therapeutic targets for the effective treatment of acute soft tissue injury.

Keywords: Exosomes; Gene Ontology; MicroRNAs; Sequencing; Soft Tissue Injuries.