[Effect of BMSCs transplantation on VEGF receptor fetal liver kinase 1 after spinal cord injury in rats].
ABSTRACT To observe the effect of BMSCs transplantation on gene and protein expression of VEGF receptor fetal liver kinase 1 (Flk-1) after spinal cord injury (SCI), and to investigate the mechanism of repairing the SCI by BMSCs transplantation.
BMSCs were isolated and cultured from five 4-week-old male Wistar rats weighing 100-120 g. The SCI model was made by using the modified Allen's impactor device. Eighty-one adult female Wistar rats weighing 220-250 g were randomly divided into 3 groups: sham-operated group (group A, n=21), in which spinous process and vertebral plate of thorax 8-10 spinal cord segment were removed; DMEM group (group B, n=30), in which rats received four injections of DMEM in the peri-lesion area; and BMSCs group (group C, n=30), in which rats received four injections of BMSCs in the peri-lesion area. The changes of Flk-1 mRNA expression in rats' spinal cord tissues were detected with RT-PCR method 1, 3 and 5 days after transplantation. The expression of Flk-1 protein was observed by using immunohistochemical technology in spinal cord 3, 7 and 14 days after transplantation.
Morphology of the primary cultured BMSCs was various. Cell morphology tended to be uniform with the accumulation of passages, which appeared flat and spindle-shaped. RT-PCR results showed that there was no significant differences (P> 0.05) in Flk-1 mRNA expression between group C and group B at different time points after transplantation. But Flk-1 mRNA levels of group B and group C significantly increased and peaked 1 day after transplantation (P < 0.01), and then decreased 3 days after transplantation (P < 0.01) compared with that of group A, and were still higher than that of group A 5 days after transplantation (P < 0.05). Immunohistochemical staining results revealed that the expression of Flk- 1 in group B was enhanced 3 and 7 days after transplantation compared with group A, which was significantly different (P < 0.01). There was no significant difference in the expression of Flk-1 between group B and group A 14 days after transplantation (P > 0.05). There was no significant difference in Flk-1 protein expression between group C and group B 3 days after transplantation (P > 0.05). The expression of Flk-1 protein in group C was significantly higher than that in group B 7 and 14 days after transplantation (P < 0.01).
BMSCs transplantation after SCI does not have regulatory effect on the expression of Flk- 1 mRNA, but it does upregulate the Flk-1 protein expression, which may be one of the mechanisms of repairing SCI.