GDNF基因修饰的神经干细胞促进脑中风后大鼠的GAP-43表达

The Grafting Neural Stem Cells Modified by GDNF Gene Enhances the Expression of GAP-43 in Rats Subjected to Cerebral Ischemia Reperfusion

目的研究胶质源性神经营养因子(GDNF)基因修饰的神经干细胞(NSCs)移植对脑中风后大鼠缺血侧脑组织内生长相关蛋白-43(GAP-43)表达的影响,探讨GDNF基因修饰的神经干细胞(GDNF/NSCs)移植对大鼠脑中风的神经保护作用机制。方法取新生大鼠脑组织分离培养NSCs,收集第6代前的NSCs备用。用重组腺病毒GDNF转染神经干细胞,制备GDNF/NSCs。暂时性阻塞大鼠大脑中动脉制备脑中风模型,3天后,用脑立体定位仪向中风侧侧脑室分别给予NSCs、GDNF/NSCs和生理盐水。再灌注时间1w、2w、3w、5w、7w后处死大鼠(n=3)。裂解中风侧脑组织,离心后得到脑组织蛋白样品,通过蛋白免疫印迹(Western Blotting)检测GAP-43表达。结果 GDNF/NSCs、NSCs、NS各组GAP-43的表达在1w、2w、3w、5w、7w各时间点均逐渐降低。NSCs组、GDNF/NSCs组显著高于NS组(P<0.05);GDNF/NSCs组高于NSCs组(P<0.05)。结论 GDNF/NSCs移植治疗脑中风的机制可能与促进GAP-43表达有关。

Objective To study the effects of grafting neural stem cells( NSCs) modified by glial cell line-derived neural factor( GDNF) gene ( GDNF/NSCs) on the expression of GAP-43 in rats subjected to cerebral ischemia reperfusion. Methods NSCs were cultured from newborn rats and NSCs before 6 generation were used for grafting use. NSCs were infected by recombinant GDNF adenovirus to prepare NSCs-overexpressing GDNF( GDNF/NSCs) . Rat stroke was performed by occluding the middle cere-bral artery occlusion for 2 h and reperfusion. At 3 days after reperfusion,NSCs,GDNF/NSCs and saline was infused into ipsilateral ventricle respectively. According to different reperfusion time,each group was subdivide into 5 groups:1,2,3,5,7 weeks(n=3). At each time point,rats were sacrificed and brains were removed. Detect the expression of GAP-43 by Western Blotting. Results The expressions of GAP-43 in all the three groups gradually were decreased at given time point. NSCs group and GDNF/NSCs group significantly increased GAP-43 compared with ischemic group(P<0. 05). Besides,The GDNF/NSCs group showed more ef-ficient than NSCs group(P<0. 01). Conclusion Neuroprotection of the grafting GDNF/NSCs may be realted to enhance the ex-pression of GAP-43.