Difference In The Expression Of Notch-1 Gene During Muscle-derived Stem Cells Repairing Spinal Cord Injury In Rats

ObjectiveTo dicuss the expression difference of Notch1 in the process of pepairing the injure of the rats'spinal cord by the muscle derived stem cells.Methods1.Separate and bring up rats'MDSCs by the method of mechanical mixing enzyme digestion.2.Seperate the rats by random number table:NumberA:the MDSCs after induced by NGF and BrdU the group of treatment(n=30);NumberB: the MDSCs which was't induced but BrdU the group of treatment(n=30),Number C:the group of contro(ln=30)and the group of false surgery(n=30).The group A.B.C was divided to one week four weeks eight weeks by the different time of choosing materials.3.The surgery method:make the the cross-sectional model of the spinal cord.After one week of the damaging of the spinal cord,inject the MDSCs medium which had been induced by NGF and BrdU group A with microscale injector.Inject the equal volume MDSCs medium which was't induced but BrdUto group B.Inject the equal volume PBS medium to groupD. 4.Determine the sport fuction of the hind legs of each rat in different time when they are alive.(BBB sport function score and inclined plane test).5.Detect and observed the survival.migration and differentiation of MDSCs in the rats by the pathological detection and immune and staining by histochemical.6.Detect the expression of the key gene and protein in the Notch1 after planting the MDSCs by RT-PCR and Western. Results1.After the machine-mixed enzyme digestion and differential adhesion, we succeed to foster high purity, high activity MDSCs, immunocytochemistry results is Desmin (+) (positive rate> 80%).2.In the process of inducing, the cells grew well and showed a gradual change in their shape, and no neurospheres was formed;After a week, the NSE staining of the bodies and apophysis of nerve-like cell was strongly positive (37.29±1.59)%, rate of cell positive cells which was't induced was low (4.26±0.34)%, the difference between the two groups was significantly (P <0.01); The expression level of NSE in the sample MDSCs is higher than the uninduced group (P <0.01).3.Behavior observations: after 1 week of their injury, there wsn't significant difference by BBB scores and inclined board test (P <0.01)in each group, with the time going.The differences between each group became clear, scores of group A (induced group) was significantly higher than group B (no induction group), and was significantly better than the group C (control group),which wsn't interevented. It showed that treatment of spinal cord injury by MDSCs which is induced by NGF is significantly better than the single application of the transplantation methods of MDSCs.4.Histological results: aetect and the survival. migration and differentiation of the planted MDSCs in each group. And MDSCs in each group all migrated the surrounding area of spinal cord injury, there was only a little existed in the gray matter. Induced damage and repair plant group is particularly evident than cells in group which was't induced. The results showed that the effect of MDSCs of repairing the spinal cord injury was better than the single application of MDSCs.5.RT-PCR results showed that: The mRNA expression levels of Notch1 in Notch signaling pathway in Notch1 in the induction group and non-induction group were significantly different (P<0.01), the degree of molecules expression in Notch pathway signaling in the process of inducing differentation was significantly reduced. Conclusions1.DSCs can be cultured in vitro and can be identified by Desmin stain and used to tissue engineering as"seed"cells.2.Ith the aid of NGF from MDSCs into spinal cord injury of the rats, and achieved certain results, which MDSCs as seed cells to repair spinal cord injury is important.3.Otch signaling pathway played an important role in the process of MDSCs induced into neural-like cells. Low expression of Notch signaling molecules can promote the MDSCs differentiated into neural-like cells.