| Objective:By in vitro experiments designed to observe the low frequency weak magnetic neural stem cells in bone marrow transplantation for brain injury rats crawling function improved.Observed related markers and neurotransmitters neurons expression changes,provides the basic research basis for the clinical treatment of brain injury.Methods:A total of 64 SD rats were randomly divided into four groups:control group(group C),BM-NPCs group(group B),magnetic field group(group L)and group B Magnetic field + BM-NPCs group(LB group).Bone Marrow Stroma Cells-derived Neural Progenitor Cells(BM-NPCs)were cultured in bone marrow-derived bone marrow mesenchymal stem cells from 3-week-old SD rats.On the 7th day after successful modeling,CD-Dil Marked BM-NPCs.Among them,L group and LB group were low frequency weak magnetic field(50Hz,5mT)intervention,60min/d,continuous intervention 60d.In the course of the experiment,the rats were sacrificed at 1d,3d,7d,30d and 60d,and the Walkne clark was graded.The rats were subjected to HE staining at 1d,7d and 30d,The expression of NeuN,GFAP and ?-IIItubulin in the transplanted cells was detected by immunofluorescence in the wound area.In vitro experiments,the use of 50 Hz,5 mT weak magnetic field to deal with BM-NPCs,time for 60 min/day,15 days continuous action,at the same time set up the control group.The expression of Nestin(neural stem cell marker),Tuj-1(neuron marker)and GFAP(glial cell marker)in BM-NPCs were detected by immunofluorescence assay.The expression of BM-NPCs was detected by Q-PCR.(Neural stem cell markers),PSA-NCAM,?-IIItubulin,ACHE,5-HT,GABA(neurotransmitters)and the like.The results of:There was no significant difference between the two groups(P>0.05).The scores of Wayne clark were observed at the 3rd,7th,30th and 60th day after transplantation,and the results were not significantly different between the two groups(P>0.05)(P<0.05).There were significant differences between the two groups in the C group and the other three groups.There were differences between the two groups at different time points,and the scores of different treatment factors increased with time The trend of different groups,which LB group motor function score improvement is more obvious.The results of HE staining showed that the vascular tissue was deformed and the swelling and necrosis of the tissue were observed on the 7th day after the model.BM-NPCs on the 7th day after transplantation,the edema of the brain tissue around the brain injury group C and the L group was significantly reduced,there were larger cystic cavities,infiltration of inflammatory cells and significant reduction of nerve cells.B group and LB group Edema,the scope of the limitations of the cystic cavity,visible glial cells.On the 30th day after transplantation,the other three groups had less cystic cavity area,and the edema tissue in group B disappeared.The cells around the brain tissue of LB group were neat and the inflammatory cells disappeared.At 60 days after transplantation,compared with the control group,the brain tissue of most experimental groups was repaired.Dil-labeled NeuN and ?-IIItubulin-positive cells were found in the brain tissue of the B and LB rats on the 30th day after BM-NPCs transplantation.On the 60th day after transplantation,the Dil-labeled NeuN And?-IIItubulin positive cells and brain tissue around the brain fusion growth.A large number of GFAP-positive cells were found in the brain tissue of the rats in group C on the 30th day and the 60th day after BM-NPCs transplantation,while the L group and the LB group were less than the C group.In the course of in vitro experiments,BM-NPCs cultured in vitro formed cell spheres and treated with weak magnetic field,neuronal-like cells were observed.The expression of Nestin in BM-NPCs was positive in immunocytochemistry.The expression of Tuj-1 and GFAP in the control group and the magnetic field group were positive.Compared with the control group,Q-PCR results showed that the expression levels of Nestin,PSA-NCAM,Ach,GABA and 5-HT in the magnetic field group were significantly different from those before induction(P<0.01)MRNA expression was significantly higher than that in the control group(P<0.05).Conclusion:Low-frequency weak magnetic energy can improve the ipsilateral crawling function of brain injury in rats,and can also promote the repair of bone marrow-derived neural stem cells in rat brain injury.The process and the low-frequency weak magnetic field up-regulate the expression of ?-? tubulin mRNA to promote bone marrow stem cells Cell differentiation. |
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