ATRA干预培养鼠胚神经干细胞联合GDNF移植对脊髓损伤再生修复的作用

发布时间：2018-07-04 10:34

本文选题：全反式维甲酸 + 神经干细胞　；参考：《山东医药》2014年22期

【摘要】：目的体外应用全反式维甲酸(ATRA)进行干预培养的鼠胚神经干细胞(NSCs)联合胶质细胞源性神经营养因子(GDNF)移植至大鼠损伤脊髓,探讨其对脊髓损伤再生修复的效果。方法体外应用ATRA 1.0μmol/L浓度下诱导NSCs,将60只雌性SD大鼠随机分为正常组、假手术组、损伤对照组、ATRA干预组、GDNF组、ATRA+GDNF组,每组10只。损伤对照组、ATRA干预组、ATRA+GDNF组、GDNF组大鼠行胸段脊髓全横断损伤效果。术后于移植前1 d、移植后1、2、5、8周,评估大鼠双后肢运动功能,测定运动诱发电位(MEP)评估神经传导功能,采用免疫荧光染色观察移植后NSCs存活情况,5-溴脱氧尿嘧啶核苷/胶质纤维酸性蛋白(BrdU/GFAP)免疫荧光双标染色观察星形胶质细胞分化情况,5-溴脱氧尿嘧啶核苷/微管相关蛋白-2(BrdU/MAP-2)免疫荧光双标染色观察神经元分化情况。评估轴突生长情况。结果 8周后ATRA+GDNF组大鼠行为学评分高于其他各组(P0.01)。与ATRA干预组及GDNF组相比,8周后ATRA+GDNF组脊髓内GFAP、MAP-2阳性细胞表达面积增大(P0.05);移植的NSCs在受损脊髓内存活并向周围迁移,后肢运动功能较对照组及ATRA干预组、GDNF组明显改善。结论体外ATRA干预的NSCs联合GDNF移植可有效补充缺失的神经细胞,改善脊髓内微环境,对大鼠后肢功能的改善优于单一应用细胞或神经营养因子移植。
[Abstract]:Objective to investigate the effect of rat embryonic neural stem cells (NSCs) combined with glial cell derived neurotrophic factor (GDNF) on the regeneration and repair of spinal cord injury (sci) after intervention with all-trans retinoic acid (ATRA) in vitro. Methods NSCs were induced by ATRA 1.0 渭 mol / L in vitro. Sixty female SD rats were randomly divided into normal group, sham-operated group and ATRA GDNF group with 10 rats in each group. The rats in ATRA GDNF group received thoracic spinal cord transection. Motor function of the hind limbs and motor evoked potential (MEP) were evaluated 1 day before transplantation and 8 weeks after transplantation, and the nerve conduction function was evaluated by motor evoked potential (MEP). Observation of survival of NSCs after transplantation by immunofluorescence staining using 5-bromodeoxyuridine / glial fibrillary acidic protein (BrdU / GFAP) immunofluorescence double labeling staining to observe the differentiation of astrocytes with 5-bromodeoxyuridine / microtubule phase BrdU / MAP-2 (BrdU / MAP-2) immunofluorescence double labeling staining was used to observe the differentiation of neurons. Evaluate axon growth. Results after 8 weeks, the behavioral score of ATRA GDNF group was higher than that of other groups (P 0.01). Compared with ATRA intervention group and GDNF group, the area of GFAP-MAP-2 positive cells in the spinal cord of ATRA GDNF group increased after 8 weeks (P0.05), the transplanted NSCs survived and migrated around the injured spinal cord, and the motor function of hind limbs was significantly improved compared with the control group and the ATRA intervention group. Conclusion in vitro ATRA combined with GDNF transplantation can effectively supplement the missing nerve cells and improve the microenvironment of spinal cord. The improvement of hind limb function of rats is better than that of single cell or neurotrophic factor transplantation.
【作者单位】：泸州医学院研究生院;泸州医学院附属医院;
【基金】：四川省卫生厅科研资助课题(2008-527-080178) 泸州市重点科技计划资助项目(2008-23-5)
【分类号】：R651.2

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