摘要
用含2.4 mg.mL-1川芎嗪提取液对小鼠骨髓间充质干细胞(BMSCs)进行诱导,探讨川芎嗪体外诱导BMSCs分化为神经元样细胞的作用.应用EGTA(细胞外Ca2+螯合剂)、Nifedipine(L-型Ca2+通道阻断剂)和LY294002(PI3K阻断剂)等Ca2+阻断剂分别作用细胞,RT-PCR和Western blot技术研究Ca2+信号在川芎嗪诱导BMSCs分化为神经细胞过程中的作用.结果表明:川芎嗪作用不同时间的BMSCs均可见Nestin、β-Tubulin Ⅲ、NSE和Nurrl的表达;川芎嗪诱导后的细胞浆内Nestin和NSE蛋白表达呈阳性.EGTA、Nifedipine及LY294002分别阻断细胞外Ca2+、L-型Ca2+通道及PI3K后,NSE和Nurr1基因及NSE蛋白表达较川芎嗪诱导组显著上调.以上结果说明川芎嗪能使BMSCs定向分化为神经元样细胞,细胞内、外Ca2+的减少可促进川芎嗪诱导BMSCs向神经细胞的分化,Ca2+信号在川芎嗪诱导BMSCs向神经细胞定向分化过程中起负调控作用.
Mouse bone marrow-derived mesenchymal stem cells（BMSCs） were induced by the induction medium containing 2.4 mg·mL-1 tetramethylpyrazine to investigate the ability of BMSCs to differentiate into neuron-like cells in vitro,and the blocking agents,such as EGTA（Ca2＋ chelator）,Nifedipine（L-type Ca2＋ channel blocker）and LY294002（PI3K inhibitor）,were applied to block the cellular Ca2＋ signal pathway to further study the effect of Ca2＋ signal on the neural differentiation of BMSCs induced by tetramethylpyrazine with RT-PCR and Western blot methods.RT-PCR analysis revealed that an expression of several neural genes including Nestin,β-Tubulin III,NSE and Nurrl were found in the induced BMSCs.The expression of several neural specific proteins,such as Nestin and neuron-specific enolase（NSE） were detected by Western blot method.NSE and Nurr1 were conspicuously up-regulated after the Ecto Ca2＋,L-type Ca2＋ channel and PI3K blocked respectively by EGTA,Nifedipine and LY294002.These results suggested that tetramethylpyrazine can induce the BMSCs differentiate into nerve cells,the reduction of ectocellular or intracellar Ca2＋ can contribute to neural differentiation,so the Ca2＋ signaling pathway play an important negative regulation role in the neural differentiation of BMSCs induced by tetramethylpyrazine.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2010年第2期1-5,9,共6页
Journal of Gansu Agricultural University
基金
甘肃省自然科学基金(3ZS051-A25-078)
