三种补肾中药有效成分对AD小鼠胚胎神经干细胞自我更新及神经元样分化作用研究  

Effective Components of Three Kinds of Shen-supplementing Chinese Medicine on Self-renewal and Neuron-like Differentiation of NSCs in AD Mouse Embryos:an Experimental Research

作　　者：张玉莲[1] 张琳琳[1] 宋宛珊[2] 韩文文[1] 黄建华[3] 周震[1] 

机构地区：[1]天津中医药大学第二附属医院脑病科,天津300150 [2]天津中医药大学研究生院,天津300193 [3]复旦大学华山医院中西医结合研究所,上海200040

出　　处：《中国中西医结合杂志》2014年第10期1245-1249,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家重点基础研究发展计划(973计划)(No.2010CB530405);国家自然科学基金资助项目(No.81202654)

摘　　要：目的观察三种补肾中药补骨脂、女贞子、何首乌各自有效成分补骨脂素、齐墩果酸、二苯乙烯苷对Aβ25-35介导的神经干细胞(neural stem cells,NSCs)自我更新及向神经元方向分化的调控作用。方法取孕14天昆明小鼠体外分离、培养其胚胎神经干细胞,并随机分为对照组、Aβ25-35组、Aβ25-35+补骨脂素组、Aβ25-35+齐墩果酸组、Aβ25-35+二苯乙烯苷组,其中Aβ25-35干预浓度为25μmol/L,中药有效成分的干预浓度均为10-7mol/L,通过NSCs计数法观察NSCs增殖能力;采用Western blot及免疫荧光法观察神经元标志物Tubulin蛋白表达。计算Tubulin+/DAPI比值。结果与对照组比较,Aβ25-35组NSCs球的形态破坏,NSCs计数、Tubulin蛋白表达及Tubulin+/DAPI均降低(P<0.01,P<0.05)。与Aβ25-35组比较,3个给药组NSCs计数、Tubulin蛋白表达及Tubulin+/DAPI均升高(P<0.01,P<0.05)。结论 25μmol/L Aβ25-35可抑制NSCs自我更新及神经元样分化,而补骨脂素、二苯乙烯苷及齐墩果酸能促进NSCs的自我更新,并能促进其向神经元方向分化。Objective To observe the regulatory effects of psoralen, oleanolic acid, and stiibene glucoside, three active components of psoralea fruit, glossy privet fruit and tuber fleeceflower root respectively, on AI325_ssinduced self-renewal and neuron-like differentiation of neural stem cells （NSCs）. Methods Embryonic NSCs were in vitro isolated and cultured from Kunming mice of 14-day pregnancy, and randomly divided into the control group, the Aβ25-35 group, the Aβ25-35 ＋ psoralen group, the Aβ25-35 ＋ oleanolic acid group, and the Aβ25-35 ＋stilbene glucoside group. The intervention concentration of Aβ25-35 was 25 μmol/L, and the intervention concentration of three active components of Chinese medicine was 10^-7mol/L. The effect of three active components of Chinese medicine on the proliferation of NSCs was observed by counting method. The protein expression of Tubulin was observed by Western blot and immunofluorescence. The ratio of Tubulin ^＋/DAPI was caculated. Results Compared with the control group, the sperical morphology of NSCs was destroyed in the Aβ25-35 group, the counting of NSCs, the expression of Tubulin protein, and the ratio of Tubulin^＋/DAPI all decreased （P 〈0.01, P 〈0.05）. Compared with the Aβ25-35 group, the counting of NSCs, the expression of Tubulin protein, and the ratio of Tubulin^＋/DAPI all increased in the three Chinese medicine treated groups （P 〈0.01, P 〈0.05）. Conclusions 25 μmol/LAβ25-35 could inhibit self-renewal and neuron-like differentiating of NSCs. But psoralen, oleanolic acid, and stilbene glucoside could promote self-renewal of NSCs and neuron-like differentiation.

关 键 词：补肾中药有效成分 Β淀粉样蛋白 神经干细胞 神经元 调控作用 

分 类 号：R285[医药卫生—中药学]