Low-intensity pulsed ultrasound regulates proliferation and differentiation of neural stem cells through notch signaling pathway
Low-intensity pulsed ultrasound regulates proliferation and differentiation of neural stem cells through notch signaling pathway
复制标题
低强度脉冲超声通过Notch信号通路调控神经干细胞增殖和分化
DOI：
10.1016/j.bbrc.2020.03.142
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发表时间：
2020-06-04
影响因子：
3.1
通讯作者：
Feng, Shiqing
中科院分区：
文献类型：
--
作者：
Wu, Yu;Gao, Qiang;Feng, Shiqing
低强度脉冲超声（LIPUS）被广泛用于调节干细胞增殖和分化。然而，LIPUS刺激对神经干细胞（NSC）的影响还没有很好的记录。在本研究中，我们已经确定了最佳的参数，并探讨了LIPUS在体外调控神经干细胞增殖和分化的细胞机制。获得神经干细胞并通过巢蛋白免疫染色进行鉴定。CCK-8法检测神经干细胞增殖情况。采用免疫分析法（ELISA）检测各组大鼠血清中营养因子（NTFs）的表达。免疫荧光和免疫印迹法检测神经干细胞的分化。还通过免疫印迹测定来测量参与Notch信号传导途径的蛋白质的表达水平。结果表明，69.3mW/cm（2）（1 MHz，8V）的强度适合于LIPUS刺激。ELISA分析表明，LIPUS处理促进了体外培养的神经干细胞营养因子的表达。免疫荧光和免疫印迹分析表明，LIPUS不仅减少星形胶质细胞的分化，但也刺激分化为神经元。此外，LIPUS刺激显著上调Notch 1和Hes 1的表达水平。我们的研究结果表明，LIPUS通过调节Notch信号通路触发NSCs增殖和分化。这项研究表明，LIPUS是一个潜在的和有前途的治疗平台，用于优化干细胞，并使中枢神经系统疾病的非侵入性神经调节成为可能。(C)2020爱思唯尔公司All rights reserved.
Low-intensity pulsed ultrasound (LIPUS) is widely used to regulate stem cell proliferation and differentiation. However, the effect of LIPUS stimulation on neural stem cells (NSCs) is not well documented. In this study, we have identified the optimal parameters, and investigated the cellular mechanisms of LIPUS to regulate the proliferation and differentiation of NSCs in vitro. NSCs were obtained and identified by nestin immunostaining. The proliferation of NSCs were measured by using Cell Counting Kit-8 (CCK-8). The expressions of nutritional factors (NTFs) were detected with immunoassay (ELISA). NSCs differentiation were detected by immunofluorescence and immunoblotting analysis. The expression level of proteins involved in the Notch signaling pathway was also measured by immunoblotting assay. Our results showed the intensity of 69.3 mW/cm(2) (1 MHz, 8 V) was applicable for LIPUS stimulation. ELISA analysis demonstrated that LIPUS treatment promoted the expression of nutritional factors of NSCs in vitro. Immunofluorescence and immunoblotting analyses suggested that the LIPUS not only reduced the astrocyte differentiation, but also stimulated the differentiation to neurons. Additionally, LIPUS stimulation significantly upregulated expression level of Notch1 and Hes1. Results from our study suggest that LIPUS triggers NSCs proliferation and differentiation by modulating the Notch signaling pathway. This study implies LIPUS as a potential and promising therapeutic platform for the optimization of stem cells and enable noninvasive neuromodulation for central nervous system diseases. (C) 2020 Elsevier Inc. All rights reserved.