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

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Mashl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Mashl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20 mg/L剂量最明显,差异均有统计学意义(P<0.05);20 mg/L组Mashl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,能够促进神经干细胞中Mashl的表达.

Effect of Tenuigenin on Mash1 expression and directional differentiation of neural stem cells from mouse hippocampus in vitro

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 Mash1 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 Mash1 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.