Bcl-2 gene-modified neural stem cell transplantation for spinal cord injury in rats
10.3969.j.issn.1671-7856.2016.07.006
- VernacularTitle:bcl-2基因修饰神经干细胞移植修复大鼠脊髓损伤
- Author:
Mei ZHANG
;
Yuexin WANG
;
Xiaohua HOU
;
Jun HONG
;
Shengchun YIN
;
Yan LI
;
Qingyang LIU
- Publication Type:Journal Article
- Keywords:
Spinal cord injury;
Bcl-2 gene;
Modification;
Neural stem cells;
Transplantation;
Repair;
Rat;
Nerve function
- From:
Chinese Journal of Comparative Medicine
2016;26(7):35-41
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the bcl?2 gene modification on neurological function recovery in rats with spinal cord injury in neural stem cell transplantation. Methods Cultured rat neural stem cells by Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene ( bcl?2 ) gene transfection of neural stem cells were divided into 3 groups: control group, negative transfection group, bcl?2 transfection group. Use western?blot to detect the expression of bcl?2 protein in neural stem cells before and after transfection. 85 adult female SD rats, successful model 72, were randomly divided into control group, NSCs group, bcl?2?NSCs groups, 24/group, rat acute spinal cord injury model in accordance with a modified Allen’ s method. Assess the motor function by BBB rating and the swash plate test. 7 days after modeling by RT?PCR and Western blot detection of spinal cord injury around HSP27, c?fos gene expression, TUNEL assay apoptosis. Four weeks after model drawn line HE staining and fluorescence microscopy EGFP?labeled NSC survival and distribution of the rats neurophysiological recovery by SEP and MEP. Results bcl?2 gene transfection of rat neural stem cells, bcl?2 transfection group and control group, negative transfection group compared to bcl?2 mRNA and protein levels were expressed ( P < 0. 05 ); lower extremity motor function in rats evaluation of bcl?2?NSCs group than NSCs group, NSCs group than the control group. 72 hours after modeling, bcl?2?NSCs number of apoptotic cells were significantly lower than the control group and NSCs group (P < 0. 05). 7 days after modeling, compared with the control group and NSCs group, bcl?2?NSCs group HSP27 gene and protein expression was significantly higher than that (P < 0. 05), bcl?2?NSCs group c?fos mRNA and protein expression was significantly reduced compared (P < 0. 05). 4 weeks after modeling, HE staining control group showed spinal cord tissue loss and the formation of syringomyelia, no axonal through. NSCs group damage zone few of neuraxis?like structures, syringomyelia smaller, bcl?2?NSCs group showed more nerve axon?like structure, no syringomyelia. EGFP?positive cells labeled:bcl?2?NSCs group the most, NSCs group followed, no control group, and the difference between the groups was statistically significant (P < 0. 05). After the 4th week, SEP and MEP latency period:bcl?2?NSCs group NSCs group > control group, and between the groups was significant difference ( P < 0. 05 ) . Conclusions By Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene (bcl?2) gene transfection make neural stem cells can promote cultured rat neural stem cells. bcl?2 gene?modified neural stem cell transplantation can promote the regeneration of spinal cord injury synaptic elevated HSP27 expression after spinal cord injury, reduced expression and neural cell apoptosis after spinal cord injury bcl?2 gene and improve limb movement in rats function and electrophysiological function.
