Knockdown of miR-429 Attenuates Aβ-Induced Neuronal Damage by Targeting SOX2 and BCL2 in Mouse Cortical Neurons
Knockdown of miR-429 Attenuates Aβ-Induced Neuronal Damage by Targeting SOX2 and BCL2 in Mouse Cortical Neurons
复制标题
miR-429 的敲低通过靶向小鼠皮质神经元中的 SOX2 和 BCL2 减轻 Aβ 诱导的神经元损伤
DOI：
10.1007/s11064-018-2643-3
复制
发表时间：
2018-12-01
影响因子：
4.4
通讯作者：
Zhang, Shuling
中科院分区：
文献类型：
--
作者：
Fu, Shengqi;Zhang, Jiewen;Zhang, Shuling
淀粉样肽(A)的积累和细胞凋亡导致的大量神经元死亡是阿尔茨海默病（AD）发病的重要步骤。据报道，MiR-429在AD的发病机制中发挥重要作用。然而，miR-429在AD发病机制中的详细功能和潜在分子机制尚不清楚。用20 μ M的A（25-35）刺激皮质神经元24h，体外构建AD模型。采用qRT-PCR法检测miR-429的表达，并采用qRT-PCR或western blot分析评估AD小鼠模型和a诱导的皮质神经元中性别决定区Y-box 2 （SOX2）和B细胞淋巴瘤-2蛋白（BCL2）的mRNA或蛋白水平。荧光素酶报告基因检测和western blot分析证实miR-429的潜在靶点。采用CCK-8法、流式细胞术和caspase3活性法分别测定细胞活力、细胞凋亡能力和caspase3活性。在AD小鼠模型和a诱导的小鼠皮质神经元中，MiR-429上调，SOX2和BCL2下调。MiR-429敲低可减弱a诱导的小鼠皮质神经元细胞毒性。SOX2和BCL2是miR-429的直接靶点。此外，SOX2或BCL2表达的恢复减弱了抗mir -429介导的神经保护作用。miR-429的敲低可能通过靶向小鼠皮质神经元中的SOX2和BCL2来减弱a诱导的细胞毒性，为阿尔茨海默病的治疗提供了新的前景。
Accumulation of amyloid- peptide (A) and massive neuronal death due to apoptosis were the essential steps in the pathogenesis of Alzheimer's disease (AD). MiR-429 was reported to play an important role in the pathogenesis of AD. However, the detailed function and underlying molecular mechanism of miR-429 in the pathogenesis of AD remain elusive. Cortical neurons were stimulated with 20 mu M of A(25-35) for 24h to construct AD model in vitro. qRT-PCR assay was used to detect the expression of miR-429, and qRT-PCR or western blot analysis were performed to assess the levels of Sex-determining region Y-box 2 (SOX2) and B cell lymphoma-2 protein (BCL2) at mRNA or proteins levels in the AD mouse model and A-induced treated cortical neurons. Luciferase reporter assay and western blot analysis were used to confirm the potential targets of miR-429. CCK-8 assay, flow cytometry analysis, and caspase3 activity assay were used to measure cell viability, cell apoptosis capacity and caspase3 activity, respectively. MiR-429 was upregulated and SOX2 and BCL2 were downregulated in the AD mouse model and A-induced mouse cortical neurons. MiR-429 knockdown attenuated A-induced cytotoxicity in mouse cortical neurons. SOX2 and BCL2 were direct targets of miR-429. Moreover, anti-miR-429-mediated neuroprotective effect was abated by the restoration of SOX2 or BCL2 expression. Knockdown of miR-429 might attenuate A-induced cytotoxicity by targeting SOX2 and BCL2 in mouse cortical neurons, providing a novel prospect in AD therapy.