Lentivirus-mediated microRNA-124 gene-modified bone marrow mesenchymal stem cell transplantation promotes the repair of spinal cord injury in rats.
- Author:
Jia Lin SONG
1
;
Wei ZHENG
;
Wei CHEN
;
Yun QIAN
;
Yuan Ming OUYANG
;
Cun Yi FAN
Author Information
- Publication Type:Original Article
- MeSH: Animals; Blotting, Western; Bone Marrow*; Bromodeoxyuridine; Cell Count; Enzyme-Linked Immunosorbent Assay; Epoprostenol; Gangliosides; Immunohistochemistry; Intermediate Filaments; Mesenchymal Stem Cell Transplantation*; Mesenchymal Stromal Cells*; Microtubule-Associated Proteins; Phosphopyruvate Hydratase; Pyridoxal Kinase; Rats*; Real-Time Polymerase Chain Reaction; RNA, Messenger; Spinal Cord Injuries*; Spinal Cord*; Thyrotropin-Releasing Hormone
- From:Experimental & Molecular Medicine 2017;49(5):e332-
- CountryRepublic of Korea
- Language:English
- Abstract: Our study aims to explore the effects of lentivirus-mediated microRNA-124 (miR-124) gene-modified bone marrow mesenchymal stem cell (BMSC) transplantation on the repair of spinal cord injury (SCI) in rats. BMSCs were isolated from the bone marrow of rats. The target gene miR-124 was identified using a luciferase-reporter gene assay. Seventy-two rats were selected for construction of the SCI model, and the rats were randomly divided into the blank group, sham group, SCI group, negative control (NC) group, overexpressed miR-124 group and si-PDXK group. The mRNA expression of miR-124 and the mRNA and protein expression of pyridoxal kinase (PDXK) were detected by quantitative real-time polymerase chain reaction and western blotting. The locomotor capacity of the rats was evaluated using the Basso, Beattie and Bresnahan (BBB) scale. Brdu, neuron-specific enolase (NSE), neurofilament (NF) and microtubule-associated protein 2 (MAP2) were detected using immunohistochemistry. The expression levels of thyrotropin-releasing hormone (TRH), prostacyclin (PGI2) and gangliosides (GM) were measured using an enzyme-linked immunosorbent assay. PDXK was identified as the target gene of miR-124. The overexpressed miR-124 group exhibited higher miR-124 expression than the SCI, NC and si-PDXK groups. Compared with the SCI and NC groups, the PDXK expression was downregulated in the overexpressed miR-124 and si-PDXK groups, and the BBB scores were significantly increased 7, 21 and 35 days after transplantation. The double-labeled positive cell densities (Brdu+NSE/NF/MAP2) and the expression levels of TRH, PGI2 and GM in the overexpressed miR-124 group were significantly higher than those in the NC and SCI groups. These results indicated that miR-124 targeted PDXK to accelerate the differentiation of BMSCs into neurocytes and promote SCI repair.
