动物实验,神经干细胞移植 z-bo 2020-08-26 601

　　Objective: To investigate the effect of neural stem cell transplantation modified by bcl-2 gene on the recovery of neurological dysfunction in rats with spinal cord injury.

　　Method: Culture rat neural stem cells in vitro, and use Ad-EGFP as a vector to mediate B lymphoblastoma 2 gene (bcl-2) gene transfection into neural stem cells. It can be divided into 3 groups: control group and negative transfection group. , Bcl-2 transfection group. Western blot was used to detect the expression of bcl-2 protein in neural stem cells before and after transfection. The rat model of acute spinal cord injury was established by the modified Allen method, and the rats were randomly divided into control group, NSC group, and bcl-2-NSC group with 85 adult female SD rats, a total of 72 rats. , Divided into 24 rats/group. Evaluate motor function through BBB score and tilt board test. Seven days after modeling, RT-PCR and Western blot were used to detect HSP27 and c-fos gene expression around the spinal cord injury area, and TUNEL method was used to detect cell apoptosis. Four weeks after the establishment of the model, samples were taken for HE staining of pathological sections and fluorescence microscopy to observe the survival and distribution of EGFP-labeled NSC, and to observe the neuroelectrophysiological recovery of rats through SEP and MEP.

　　Result: After the bcl-2 gene was transfected into rat neural stem cells, the bcl-2 gene and protein levels were higher than the control group and the negative transfection group (PNSC group → control group). Expression in transfection group. The difference between the two groups is significant (P\u003c0.05).

　　Conclusion: Ad-EGFP is used as a vector to mediate B lymphoma 2 gene (bcl-2) gene transfer. Neural stem cells can promote the proliferation of rat neural stem cells cultured in vitro. Bcl-2 gene-modified neural stem cell transplantation can promote spinal cord Injured rats’ nerve synapses regenerate and increase HSP27 in the spinal cord injury area. It can reduce the expression of nerve cells, bcl-2 gene expression and apoptosis in the spinal cord injury area, and improve the motor function and electrophysiological function of the rats’ limbs.