Moclobemide upregulated Bcl-2 expression and induced neural stem cell differentiation into serotoninergic neuron via extracellular-regulated kinase pathway
Moclobemide upregulated Bcl-2 expression and induced neural stem cell differentiation into serotoninergic neuron via extracellular-regulated kinase pathway
复制标题
DOI：
10.1038/sj.bjp.0706766
复制
发表时间：
2006-07-01
影响因子：
7.3
通讯作者：
Chang, Yuh-Lih
中科院分区：
文献类型：
--
作者：
Chiou, Shih-Hwa;Ku, Hung-Hai;Chang, Yuh-Lih
莫氯贝胺（MB）是一种选择性可逆抑制单胺氧化酶- a的抗抑郁药物。近年来的研究表明，抗抑郁药物具有促进神经发生发育和提高5-羟色胺（5- hydroxytryamine; 5-HT）神经元存活率的强效生长因子的特性。然而，MB是否具有神经保护作用或调节神经干细胞（NSCs）的增殖还有待阐明在本研究中，我们首先使用MTT（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四唑）实验证明了50 μ M MB可以提高NSCs的细胞活力。实时逆转录聚合酶链反应（RT-PCR）结果显示，MB的诱导可上调Bcl-2和Bcl-xL基因的表达。通过caspases 8和caspases 3、ELISA和末端dUTP镍端标记（TUNEL）实验，我们的数据进一步证实了50 mM mb处理的NSCs可以预防fasl诱导的细胞凋亡形态学结果也支持了MB促进NSCs树突发育和增加神经突扩张的证据。此外，我们发现MB处理通过激活细胞外调节激酶（ERK）磷酸化，增加了NSCs中Bcl-2的表达免疫荧光三染法发现，mb处理的NSCs培养第7天血清素-和map -2阳性细胞比例显著升高（P < 0.01）。此外，我们的数据支持MB治疗通过调节ERK1/2增加NSCs中血清素的功能性产生。综上所述，本研究结果支持MB可通过ERK通路上调Bcl-2表达，诱导NSCs向血清素能神经元分化。
1 Moclobemide (MB) is an antidepressant drug that selectively and reversibly inhibits monoamine oxidase-A. Recent studies have revealed that antidepressant drugs possess the characters of potent growth-promoting factors for the development of neurogenesis and improve the survival rate of serotonin (5-hydroxytrytamine; 5-HT) neurons. However, whether MB comprises neuroprotection effects or modulates the proliferation of neural stem cells (NSCs) needs to be elucidated.2 In this study, firstly, we used the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay to demonstrate that 50 mu M MB can increase the cell viability of NSCs. The result of real-time reverse transcription-polymerase chain reaction (RT-PCR) showed that the induction of MB can upregulate the gene expressions of Bcl-2 and Bcl-xL. By using caspases 8 and 3, ELISA and terminal dUTP nick-end labeling ( TUNEL) assay, our data further confirmed that 50 mM MB-treated NSCs can prevent FasL-induced apoptosis.3 The morphological findings also supported the evidence that MB can facilitate the dendritic development and increase the neurite expansion of NSCs. Moreover, we found that MB treatment increased the expression of Bcl-2 in NSCs through activating the extracellular-regulated kinase (ERK) phosphorylation.4 By using the triple-staining immunofluorescent study, the percentages of serotonin- and MAP-2-positive cells in the day 7 culture of MB-treated NSCs were significantly increased (P < 0.01). Furthermore, our data supported that MB treatment increased functional production of serotonin in NSCs via the modulation of ERK1/2. In sum, the study results support that MB can upregulate Bcl-2 expression and induce the differentiation of NSCs into serotoninergic neuron via ERK pathway.