Hesl基因沉默促进骨髓间充质干细胞向GABA能神经元分化的实验研究  

Research on the improvement of GABAergic differentiation in the bone marrow stromal cells withHesl gene silencing

作　　者：龙乾发[1] 汪平[1] 王安生[1] 韩蕊[1] 汪凯[1] 贾晨光[1] 费舟[1] 刘卫平[1] 

机构地区：[1]陕西省西安市中心医院神经外科,710032

出　　处：《中华神经外科杂志》2014年第2期186-189,共4页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金（31301216）

摘　　要：目的探索Hesl基因沉默对骨髓间充质干细胞（BMSCs）向GABA能神经元分化的影响。方法体外分离纯化2-3周的sD大鼠BMSCs，通过RNAi技术沉默BMSCs中的Hesl基因，而后条件诱导BMSCs向GABA能神经元分化，最后通过免疫细胞化学和Westernblot分析细胞分化趋向。结果Hesl沉默的BMSCs经条件诱导后，免疫细胞化学结果示GAD67和NeuN阳性细胞数分别为（35．7±2．8）％和（72．4±3．2）％，均明显多于对照组（P〈0．05）；Westernblot结果示实验组GAD67蛋白条带灰度值与B-actin的比值（GAD67／p-actin）为（0．282±0．042），NeuN／13-actin为（0．728±0．039），同样高于对照组（P〈0．01）。结论Hesl基因沉默可促进BMSCs向GABA能神经元分化。Objective To study the influence of Hesl gene silencing on the GABAergic differentiation of bone marrow mesenchymal stem cells （ BMSCs ）. Methods BMSCs were isolated from 2-3 weeks SD rats and purified in vitro, then characterized by flow eytometry. After that, Hesl gene of BMSCs was inhibited by RNAi technology, then BMSCs （control group） and Hesl gene silencing BMSCs （experimental group） were induced by conditioned medium （10 pJnol of retinoic acid, 1 mmol NS, O2-dibutyryl adenosine 3＇： 5＇-cyclic monophosphate, 2% B27） respectively. Finally, immunocytoehemistry and Western blot were employed to analyze the differentiation potential of the induced cells. Results Experimental group and control group were induced by conditioned medium for 3-7 days, immunocytochemistry results showed that the percentage of GAD67-and NeuN- positive cells were （35. 7 ± 2. 8） % and （72. 4 ± 3. 2） % in the experimental group, which were more than the control group significantly （ P 〈 O. 05 ） ; Western blot results also showed that the protein gray value ratio of GAD67 or NeuN compared to the 13-aetin in the experiment was significantly higher than the control group through Bandscan software analysis （ P 〈0. 01 ）. Conclusions Hesl gene silencing can promote BMSCs to differentiate into GABAergic neurons.

关 键 词：骨髓间充质干细胞 发状分裂相关增强子一1 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]