| Objective: To explore the effect of marrow-derived mesenchymal stem cells(BMSCs) transplantation in combination with repeated intraperitoneal injection of Erythropoietin(EPO) on the repair of spinal cord injury after acute spinal cord injury in rats.Methods:Using the method of density gradient centrifugation to get rat BMSCs.The cell phenotype was detected by flow cytometry, and the cells were identified by osteogenic induction and adipogenic differentiation. Establishing the model of spinal cord injury(SCI) in SD rats by compressive clamp on thoracic 10 spinal cord.80 adult healthy male Sprague-Dawley rats were selected and divided into 4groups randomly.Group A was conrtol group,group B was BMSCs transplantation group, group C was EPO injection group, group D was BMSCs transplantation combined with EPO group.Each group randomly assigned 20 rats. 5 rats in each group were selected randomly in 1d before surgery,1d after surgery,3d after surgery,7d after surgery,14 d after surgery,21 d after surgery and 28 d after surgery.each in check, before surgery and after surgery 1d,3d,7d,14 d,21d and 28 d to evaluate the motor function of rats with BBB(Bassl, Beatle, Bresnahan) score assessment of motor function.Removed the injured spinal cord tissue on 28 d after surgery to dectect the activity of NF200 with the method of immunohistochemical staining.5 rats in each group were randomly executed in 3d after surgery,7d after surgery and 14 d after surgery and took out the damage segmental spinal cord tissue to dectect the apoptosis gene expression of Caspase3 protein by Western-blot method.The obtained data were collected and analyzed statistically.Results:(1)The cells derived from density gradient centrifugation can subculture and amplification in ritro.The cell phenotype detected by flow cytometry is consistent with BMSCs and the obtained cells can be successfully induced into adipocytes and osteoblasts.It was proved that the obtained cells were rats 'BMSCs.(2)SD rats' s SCI models were successfully set up.All rats had bilateral hind limb motor dysfunction After clamp in the spinal cord,BBB score were all less than 1 point.There was no significant difference among the 4 groups.There was no significant difference of BBB between the 3d and 1w time points in each group.At each time point of2 W,3W and 4W the BBB score of group B,C and D was significantly higher than that at group A(P<0.05).The BBB score of group D was higher than that of group B and group C, it had significant statistical significance(P<0.05). There was no significant difference in BBB score beween group B group C.(3)After treatment intervention at each time point of Spinal cord injury the Caspase3 protein expression in group B group C and group D group Caspase3 was lower than the group A, the Caspase3 protein expression of group D was the lowest.(4)NF200immunohistochemical staining results showed that the NF200 positive cells in group D were significantly higher than other groups(P<0.05).Conclusion:(1)The combination of BMSCs and EPO can increase the expression of NF200 in nerve cells after spinal cord injury, and promote nerve regeneration.(2)The combination of BMSCs and EPO can improve the BBB score after spinal cord injury, and promote the recovery of spinal cord function.(3)The combination of BMSCs and EPO may reduce the apoptosis by regulating the expression of Caspase3, and promote nerve regeneration and functional repair. |
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