Journal
RNA BIOLOGY
Volume 17, Issue 10, Pages 1508-1519Publisher
TAYLOR & FRANCIS INC
DOI: 10.1080/15476286.2020.1777767
Keywords
Microrna-21; Schwann cell; axon regeneration; peripheral nerve injury
Categories
Funding
- National Natural Science Foundation of China [81571202]
- Guangdong Provincial Scientific and Technological Funds [2014B090901043]
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At present, the functional recovery after nerve injury is not satisfactory in clinical practice. The aim of this study was to explore the molecular mechanism of miR-21 promoting Schwann cells (SC) proliferation and axon regeneration after peripheral nerve injury, providing a theoretical basis for injured nerve repair. Nerve injury models were constructed to determine the expression of miR-21 in the injured nerve by Quantitative Real-Time PCR (qRT-PCR). After miR-21 over-expression SC (mimic-miR-21) group, control SC (control-miR-21) group and blank SC (RSC96) group were constructed, SC proliferation was determined by CCK-8, cell cycle was analysed by flow cytometry, dorsal root ganglion neuron (DRGn) axon regeneration was observed after DRGn was cultured with SCs for 7 days, the expressions ofTGF beta I, TIMP3, EPHA4as well as apoptosis-related proteins caspase-3 and caspase-9 were detected by qRT-PCR and Western blot in the three groups, respectively. Target genes were confirmed by dual-luciferase reporter gene assay. The expressions ofTGF beta I, TIMP3andEPHA4were assessed by immunofluorescence in vivo. qRT-PCR indicated that miR-21 expression was significantly higher in the model group than in the sham operation and blank groups. SC proliferation index (PI) was significantly higher, the apoptosis rate was significantly lower, the axon was significantly longer, and mRNA and protein expressions ofTGF beta I, TIMP3, EPHA4as well as apoptosis-related proteins caspase-3 and caspase-9 were significantly lower in the mimic-miR-21 group than in the control-miR-21 and RSC96 groups. The double luciferase assay confirmed thatTGF beta I, TIMP3andEPHA4were potential target genes of miR-21. In vivo immunofluorescence also indicated that expressions ofTGF beta I, TIMP3, EPHA4were lower in the mimic-miR-21 group than in the control-miR-21 and RSC96 groups. We conclude that during injured peripheral nerve repair, miRNA-21 plays an important role in promoting SC proliferation and axon regeneration by regulatingTGF beta I, TIMP3andEPHA4target genes.
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