红景天苷诱导骨髓间充质干细胞向神经细胞定向分化的机制研究  

Salidroside via ERK1/2 and PI3K/AKT/mTOR signal pathway induces mouse bone marrow mesenchymal stem cells differentiation into neural cells

作　　者：陈亚男[1] 刘辉[1] 赵红斌[2] 刘阳[2] 白洁[2] 朱晓娟[2] 王玉[1] 

机构地区：[1]兰州大学公共卫生学院,甘肃兰州730000 [2]兰州军区兰州总医院骨科研究所,甘肃兰州730000

出　　处：《药学学报》2013年第8期1247-1252,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上项目(81073156)

摘　　要：探讨细胞外信号调节激酶(extracellular signal-regulated kinase,ERK1/2)和PI3K/AKT/mTOR信号通路介导红景天苷诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向神经细胞定向分化中的作用。以小鼠BMSCs细胞系(D1细胞)为研究对象,实验分为对照组和红景天苷诱导组。红景天苷(5、25、50、100和200μg.mL 1)作用于细胞24 h和红景天苷(100μg.mL 1)作用细胞0.5、1、3、6、9、12、24、48和72 h。采用细胞免疫荧光染色方法、实时定量PCR(real-time PCR)和Western blotting技术分别检测细胞NSE、MAP2、β-Tubulin III、NES和GFAP的阳性率和β-Tubulin III、NSE、ERK1/2和AKT的表达;PD98059和LY294002分别阻断ERK1/2和PI3K/AKT/mTOR信号通路后,检测ERK1/2和NSE的表达。结果表明,随着红景天苷诱导时间的延长,NSE、MAP2、NES、β-Tubulin III和GFAP的阳性率明显增加,NSE和β-Tubulin III蛋白的表达显著上调(P<0.01)。随着红景天苷水平和作用时间的增加,ERK1/2、AKT mRNA和蛋白的表达明显上调;PD98059和LY294002分别阻断EKR1/2和PI3K/AKT/mTOR信号通路后,显著抑制红景天苷促进NSE、NES和β-TubulinIII mRNA和NSE蛋白表达。结果提示,红景天苷通过激活ERK1/2和PI3K/AKT/mTOR信号通路,促进BMSCs向神经细胞定向分化。To investigate the role of the extracellular signal-regulated kinase （ERK1/2） and PI3K/AKT/ roTOR signal pathway inducing bone marrow mesenchymal stem cells （BMSCs） differentiation into neural cells, mouse bone marrow-derived mesenchymal stem cell lines D1 cells were used as research object. And they were divided into control groups and salidroside （SD） groups. Different concentrations （5, 25, 50, 100 and 200 μg·mL^-1） of SD were used and SD （100 μg·mL^-1） was used to induce at different time （0.5, 1, 3, 6, 9, 12, 24, 48 and 72 h）. The immunofluorescence staining chemical technology, real-time PCR and Western blotting were used to detect the positive rates of NSE, MAP2, β-Tubulin Ⅲ, NES, GFAP and the expression levels of β-Tubulin Ⅲ, NSE, ERK1/2, AKT. The expression of ERK1/2 and NSE was detected when the ERK1/2 and PI3K/AKT/ mTOR signal pathway was blocked by PD98059 and LY294002. It indicated that the positive rates of NSE, MAP2,/%Tubulin Ⅲ, NES and GFAP were gradually enhanced with time increased. The expression level of NSE and fl-Tubulin Ⅲ protein were significantly higher than those in control groups （P 〈 0.01）. The expression ofERK1/2, AKT mRNA and protein were higher with concentration and time increased. When the ERK1/2 and PI3K/AKT/mTOR signal pathway were blocked, the expression levels of NSE, NES and β-Tubulin Ⅲ mRNA and NSE protein were inhibited significantly. It points out that SD can stimulate the ERK1/2 and PI3K/AKT/mTOR signal pathway to promote BMSCs differentiation into neural cells.

关 键 词：红景天苷 骨髓间充质干细胞 信号通路 

分 类 号：R963[医药卫生—微生物与生化药学]