| Objective:The aim of this study is to make the electro-acupuncture therapy of GDNF and HIF-1?gene modified neural stem cells transplantation act on the spinal cord injury model.Through the behavioral evaluation of spinal cord injury rats,the influence of the content of?-catenin and GFAP in the injured tissues,the therapeutic effect of this therapy on spinal cord injury was evaluated,and the electro-acupuncture therapy of GDNF and HIF-1?gene modified neural stem cells transplantation was further applied in clinical practice.Provide theoretical basis.Methods:48 adult SD rats were fed adaptively for one week under the same conditions.They were randomly divided into four groups:12 rats in normal group?group A?were not given special treatment.The remaining 36 SD rats were divided into:namely injury group?group B?,GDNF,HIF-1?gene modified neural stem cells group?group C?,Electroacupuncture plus GDNF,HIF-1?gene modified neural stem cells group?group D?.The spinal cord injury model of SD rats was made by using spinal cord injury striking device,which resulted in T122 spinal cord injury in group B,group C and group D.After successful modeling,the injury group?group B?received local injection of 10ul 0.9%NaCl injection;the GDNF and HIF-1?modified NSCs group?group C?received local injection of 10ul dual gene modified NSCs fluid;the GDNF and HIF-1a modified NSCs plus electro-acupuncture group?group D?received local injection of 10ul NSCs fluid,which was treated with bilateral Xinshu and Feishu electro-acupuncture respectively.BBB hind limb scores were scored at 4 time windows?3d,1W,2W,3W?after transplantation.The injured spinal cord was taken for HE staining,immunohistochemical staining,Western Blot and other methods after perfusion and fixation.The changes of the content of?-catenin and GFAP of neural stem cells at the transplantation site were detected,and the therapeutic effect of electro-acupuncture therapy with GDNF and HIF-1a gene modified neural stem cells transplantation on spinal cord injury was observed.Results:1.The identification of NSCs conformed to the morphological rule of NSCs:the cells grew in suspension with round shape,adhered to each other and distributed globally.After virus transfection,the cells divide symmetrically at 24 hours,grow vigorously at 48hours,and there are individual cell spheres formed by multiple cells.At 72 hours,the cells divide and grow vigorously,and the cell pellets form more.2.BBB score:At 1 week,the therapeutic effect of group C and group D was similar,After 2 weeks,the therapeutic effect of group D was better than that of group C.3.HE staining:At the early stage of spinal cord injury,the cells were sparsely arranged and disordered,with punctate intracellular hemorrhage,forming obvious holes.After treatment,local intracellular hemorrhage decreased,tissue voids became smaller,cells arranged tightly,damage was alleviated,and nerve fibers gradually recovered.4.Immunohistochemical staining:The positive expression of GFAP was yellow-brown or brown,located in the cytoplasm and irregular in shape.After spinal cord injury,with the progress of treatment,the expression of GFAP in group C began to decrease after 2 weeks,and that in group D began to decrease after 3 days.After 3 weeks,the expression of GFAP in group D was lower than that in group C,indicating that electroacupuncture could inhibit the expression of GFAP.5.Western Blot test showed that the expression of beta-catenin increased first and then decreased after spinal cord injury,increased gradually within 3 days and 1W,and decreased gradually after 1W?P<0.05?.The expression of beta-catenin in group D was significantly higher than that in group A,B and C,indicating that electro-acupuncture treatment could promote the expression of beta-catenin.Conclusion:GDNF,HIF-1?gene modified NSCs transplantation combined with electroacupuncture can treat spinal cord injury by promoting the expression of?-catenin and inhibiting the expression of GFAP. |
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