Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury

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 th...

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Main Authors: Hongchang Yang, Jing Zhou, Junlei Wang, Luoning Zhang, Quzhi Liu, Jing Luo, Hongyan Jia, Li Liu, Qiang Zhou
Format: Article
Language:English
Published: Royan Institute (ACECR), Tehran 2021-09-01
Series:Cell Journal
Subjects:
Online Access:https://celljournal.org/journal/article/fulltext/circulating-exosomal-microrna-profiles-associated-with-acute-soft-tissue-injury.pdf
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author Hongchang Yang
Jing Zhou
Junlei Wang
Luoning Zhang
Quzhi Liu
Jing Luo
Hongyan Jia
Li Liu
Qiang Zhou
author_facet Hongchang Yang
Jing Zhou
Junlei Wang
Luoning Zhang
Quzhi Liu
Jing Luo
Hongyan Jia
Li Liu
Qiang Zhou
author_sort Hongchang Yang
collection DOAJ
description 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.
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spelling doaj.art-8cb6440ae5df4ea6beed6169f75dfc512022-12-21T18:34:14ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142021-09-0123447448410.22074/cellj.2021.7275Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue InjuryHongchang Yang0Jing Zhou1Junlei Wang2Luoning Zhang3Quzhi Liu4Jing Luo5Hongyan Jia6Li Liu7Qiang Zhou8Physical Education Department, Hohai University, Nanjing, Jiangsu, ChinaDepartment of Clinical Medicine, Jiangsu Health Vocational College, Nanjing, Jiangsu, ChinaHarbor, Channel and Coastal Engineering, Hohai University, Nanjing, Jiangsu, ChinaPhysical Education Department, Hohai University, Nanjing, Jiangsu, ChinaCentre of Counseling and Psychological Services, Hohai University, Nanjing, Jiangsu, ChinaCenter for Kidney Disease, 2nd Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, ChinaPort Channel and Coastal Engineering Department, Hohai University, Nanjing, Jiangsu, ChinaThe Department of Rehabilitation, Brain Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu, ChinaPhysical Education Department, Hohai University, Nanjing, Jiangsu, ChinaObjective: 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.https://celljournal.org/journal/article/fulltext/circulating-exosomal-microrna-profiles-associated-with-acute-soft-tissue-injury.pdfexosomesgene ontologymicrornassequencingsoft tissue injuries
spellingShingle Hongchang Yang
Jing Zhou
Junlei Wang
Luoning Zhang
Quzhi Liu
Jing Luo
Hongyan Jia
Li Liu
Qiang Zhou
Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury
Cell Journal
exosomes
gene ontology
micrornas
sequencing
soft tissue injuries
title Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury
title_full Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury
title_fullStr Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury
title_full_unstemmed Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury
title_short Circulating Exosomal MicroRNA Profiles Associated with Acute Soft Tissue Injury
title_sort circulating exosomal microrna profiles associated with acute soft tissue injury
topic exosomes
gene ontology
micrornas
sequencing
soft tissue injuries
url https://celljournal.org/journal/article/fulltext/circulating-exosomal-microrna-profiles-associated-with-acute-soft-tissue-injury.pdf
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