远志总皂苷对海马神经干细胞定向分化的作用及对Mash1表达的影响  

Effect of Tenuigenin on Mashl expression and directional differentiation of neural stem cells from mouse hippocampus in vitro

作　　者：朱晓峰[1] 戚旬中[2] 时晶[1] 赵锦程[2] 田金洲[1] 

机构地区：[1]北京中医药大学东直门医院老年医学研究所,北京100700 [2]佳木斯大学神经科学研究所,佳木斯154007

出　　处：《中华神经医学杂志》2009年第12期1193-1196,1202,共5页Chinese Journal of Neuromedicine

基　　金：国家重点基础研究发展规划（2003CB51710430170963）：国家自然科学基金（30170963）

摘　　要：目的观察远志总皂苷（TEN）对体外培养海马神经干细胞向胆碱能神经元定向分化的作用，并进一步分析其对神经干细胞Mash1表达的影响。方法分离新生24h内昆明小鼠海马，悬浮培养神经干细胞，取第3代神经干细胞进行分化诱导。实验分4组：空白对照组、5mg／LTEN组、20mg／LTEN组、100mg／LTEN组。应用免疫组织化学法检测神经干细胞中神经元特异性烯醇化酶（NSE）及胆碱乙酰转移酶（CHAT）阳性细胞比率；采用RT-PCR半定量法检测各组神经干细胞Mash1的表达。结果免疫组化结果显示：与对照组比较．TEN组神经干细胞向神经元及胆碱能神经元分化的比率明显升高，以20mg／L剂量最明显，差异均有统计学意义（P〈0．05）；20mg／L组Mash1的表达（0．9426±0．07286）亦明显高于对照组（0.7141±0．04016），差异具有统计学意义（P〈0．05）。结论TEN能够促进体外培养的海马神经干细胞向神经元及胆碱能神经元定向分化，能够促进神经于细胞中Mash1的表达。Objective To determine the effect of Tenuigenin （TEN） on the Mashl expression in neural stem cells （NSCs） from mouse hippocampus and directional differentiation of its NSCs in vitro. Methods NSCs were generated from the mouse hippocampus within 24 hours after birth and cultured with EGF and bFGF （20 ng/mL, each） in a 50-mm uncoated culture flask. The third-passage NSCs were cultured with TEN medium and divided them into vehicle, 5 mg/L, 20 mg/L and 100 mg/L groups according to different TEN concentrations. Immunocytochemical techniques were employed to detect the NSE-positive neurons and ChAT-positive neurons induced by TEN, and the expression of Mashl in NSCs affected by TEN was measured by RT-PCR techniques. Results The immunocytochemical and RT-PCR results revealed that ChAT-positive neurons and NSE-positive neurons were detected with statistical significance and 20 mg/L group showed a stronger effect than other concentration groups （P〈 0.05）. The expression of Mashl in the 20 mg/L group increased as compared with that in the vehicle group （P〈0.05）. Conclusion TEN can induce NSCs in vitro differentiating into neurons and cholinergic neurons, and increase the expression of Mashl in NSCs.

关 键 词：神经干细胞 胆碱能神经元 远志总皂苷 Mash1 

分 类 号：R686[医药卫生—骨科学]