摘要
目的探讨何首乌有效成分二苯乙烯苷(TSG)对β-淀粉样肽(Aβ25-31)诱导神经干细胞(NSCs)定向分化的影响。方法从孕14d昆明小鼠体外提取并培养NSCs,分别建立起两组分化系统可溶性Aβ25-31体外诱导NSCs损伤的阿尔茨海默病(AD)细胞模型:神经元细胞分化模型M1(Aβ25-312 5μmol/L)、星形胶质细胞分化模型M2(Aβ25-315μmol/L)。TSG低、中、高剂量组(TSG浓度分别为10-8、10-7、10-6mol/L)分别干预模型M1和模型M2,采用WST-1细胞增殖及细胞毒性试剂盒检测各组NSCs细胞活性,采用免疫荧光法及蛋白印迹法观察各组GFAP、Tubulin阳性细胞率及其蛋白表达情况。结果模型M1组和模型M2组细胞活性A值较相应的对照组明显降低(P<0.05);TSG中、高剂量组A值明显高于相应的模型组(P<0.05)。与相应的对照组比较,模型M1组Tubulin阳性细胞率及其蛋白相对丰度值明显降低,模型M2组GFAP阳性细胞率及其蛋白相对丰度值明显升高(P<0.05);与相应的模型组比较,TSG高剂量组Tubulin阳性细胞率及其蛋白相对丰度值均显著升高,GFAP阳性细胞率显著降低(P<0.05)。结论高剂量的TSG具有促进Aβ25-31诱导的NSCs向神经元方向分化的趋势,抑制向星形胶质细胞的分化。
Objective To study the effect of tetrahydroxy stilbene glucoside （TSG） from Heshouwu （Polygonum Multiflorum, 何首乌） on directed differentiation of β-amyloid peptide 25-31（Aβ25-31） -induced neural stem cells （NSCs）. Methods The NSCs were extracted from 14-day pregnant Kunming mice and cultured in vitro. Two differentiation systems of NSCs-injured Alzheimer＇s disease （AD） cell models induced by Aβ25-31 in vitro, i. e. the neuronal cell differentiation model M1 （Aβ25-31 25μmol/L） and astrocyte differentiation model M2 （Aβ25-31 5μmol/ L） were established. The model M1 and model M2 were interfered with low-dose （ 10-8 mol/L）, middle-dose （ 10-7 mol/L） , and high-dose （ 10 -6 mol/L） of TSG respectively. The cell activity of NSCs was detected with WST-1 cell proliferation and cytotoxicity kit. The positive cell rates and protein expressions of glial fibrillary acidic protein （GFAP） and Tubulin were observed with immunofluorescence and Western blot. Results The A values of cell activity in model M1 and model M2 groups were significantly decreased as compared with the corresponding control groups （ P 〈 0.05 ）. The A values in all dose of TSG groups were significantly higher than those in the corresponding model groups （ P 〈 0.05 ）. Comparing with the corresponding control groups, the positive cell rate of Tubulin and the relative abundance of its protein was significantly decreased in model M1 group, but the positive cell rate of GFAP and the relative abundance of its protein was significantly increased in model M2 group （P 〈 0. 05 ）. Comparing with the corresponding model groups, the positive cell rate of Tubulin and the relative abundance of its protein was significantly increased, but the positive cell rate of GFAP was significantly decreased in TSG high-dose group （P 〈 0.05）. Conclusion High-dose TSG can stimulate the differentiation of NSCs induced by Aβ25-31 into neurons and inhibit the dif- ferentiation into astrocytes.
出处
《中医杂志》
CSCD
北大核心
2014年第4期323-327,共5页
Journal of Traditional Chinese Medicine
基金
国家重点基础研究发展计划("973"计划)资助项目(2010CB530405)
关键词
何首乌
二苯乙烯苷
阿尔茨海默病
Β-淀粉样肽
定向分化
神经元细胞
星形胶质细胞
Polygonum multiflorum
tetrahydroxy stilbene glucoside
Alzheimer＇s disease
β-amyloid peptide
directed differentiation
neuronal cells
astrocytes
