Lentivirus-mediated microRNA-124 gene-modified bone marrow mesenchymal stem cell transplantation promotes the repair of spinal cord injury in rats

다국어 초록 (Multilingual 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.

참고문헌 (Reference)

1 Santos MC, "miR-124, -128, and -137 orchestrate neural differentiation by acting on overlapping gene sets containing a highly connected transcription factor network" 34 : 220-232, 2016

2 Xu W, "miR-124 regulates neural stem cells in the treatment of spinal cord injury" 529 : 12-17, 2012

3 Cheng LC, "miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche" 12 : 399-408, 2009

4 Furuta M, "miR-124 and miR-203 are epigenetically silenced tumor-suppressive microRNAs in hepatocellular carcinoma" 31 : 766-776, 2010

5 Tosolini AP, "Viral-mediated gene therapy for spinal cord injury(SCI)from a translational neuroanatomical perspective" 11 : 743-744, 2016

6 Martino G, "The therapeutic potential of neural stem cells" 7 : 395-406, 2006

7 Wang W, "The restoration of full-thickness cartilage defects with BMSCs and TGF-beta 1 loaded PLGA/fibrin gel constructs" 31 : 8964-8973, 2010

8 "Spinal Cord Injury (SCI) facts and figures at a glance" 39 : 493-494, 2016

9 Krichevsky AM, "Specific microRNAs modulate embryonic stem cell-derived neurogenesis" 24 : 857-864, 2006

10 Harada N, "Role of prostacyclin in the development of compression trauma-induced spinal cord injury in rats" 23 : 1739-1749, 2006

11 Nakanishi K, "Responses of microRNAs 124a and 223 following spinal cord injury in mice" 48 : 192-196, 2010

12 Gao XR, "Pri-miR-124 rs531564polymorphism and colorectal cancer risk" 5 : 14818-, 2015

13 Mittenhuber G, "Phylogenetic analyses and comparative genomics of vitamin B6(pyridoxine)and pyridoxal phosphate biosynthesis pathways" 3 : 1-20, 2001

14 Zou D, "Overexpression of microRNA-124promotes the neuronal differentiation of bone marrow-derived mesenchymal stem cells" 9 : 1241-1248, 2014

15 Li F, "Neuroprotective effect of bone marrow stromal cell combination with atorvastatin in rat model of spinal cord injury" 7 : 4967-4974, 2014

16 Yan T, "Neuronal markers are expressed in human gliomas and NSE knockdown sensitizes glioblastoma cells to radiotherapy and temozolomide" 11 : 524-, 2011

17 Czarnecka J, "Neurogenic differentiation of mesenchymal stem cells induces alterations in extracellular nucleotides metabolism" 118 : 478-486, 2016

18 Yu JY, "MicroRNA miR-124regulates neurite outgrowth during neuronal differentiation" 314 : 2618-2633, 2008

19 Moreno-Navarrete JM, "Metabolomics uncovers the role of adipose tissue PDXK in adipogenesis and systemic insulin sensitivity" 59 : 822-832, 2016

20 Yamaya S, "Low-energy extracorporeal shock wave therapy promotes vascular endothelial growth factor expression and improves locomotor recovery after spinal cord injury" 121 : 1514-1525, 2014

21 Zhao Y, "Loss of microRNA-124 expression in neurons in the peri-lesion area in mice with spinal cord injury" 10 : 1147-1152, 2015

22 Kirshblum SC, "International standards for neurological classification of spinal cord injury, revised 2011" 18 : 85-99, 2012

23 Salazar DL, "Human neural stem cells differentiate and promote locomotor recovery in an early chronic spinal cord injury NOD-SCID mouse model" 5 : e12272-, 2010

24 Katoh S, "High incidence of acute traumatic spinal cord injury in a rural population in Japan in 2011 and 2012 : an epidemiological study" 52 : 264-267, 2014

25 Gao Y, "Functional regulatory roles of microRNAs in atherosclerosis" 460 : 164-171, 2016

26 Sun Z, "Electroacupuncture-enhanced differentiation of bone marrow stromal cells into neuronal cells" 18 : 398-406, 2009

27 Baykal S, "Effect of thyrotropinreleasing hormone on Na(+)-K(+)-adenosine triphosphatase activity following experimental spinal cord trauma" 36 : 296-299, 1996

28 Zhou LL, "Effect and mechanism of different ways of transplanting bone marrow mesenchymal stem cells in cardiopulmonary resuscitation in rats" 13 : 7937-7949, 2014

29 Safo MK, "Crystal structure of pyridoxal kinase from the Escherichia coli pdxK gene : implications for the classification of pyridoxal kinases" 188 : 4542-4552, 2006

30 Xu D, "Clinical study of ganglioside(GM)combined with methylprednisolone(MP)for early acute spinal injury" 28 : 701-704, 2015

31 Shichinohe H, "Bone marrow stromal cells promote neurite extension in organotypic spinal cord slice : significance for cell transplantation therapy" 22 : 447-457, 2008

32 Basso DM, "A sensitive and reliable locomotor rating scale for open field testing in rats" 12 : 1-21, 1995