三七总皂苷通过调控 DPYSL2 促进神经干细胞向神经元分化
DPYSL2 is a novel regulator for neural stem cell differentiation in rats: revealed by Panax notoginseng saponin administration.
机构信息
Institute of Neurological Disease, Translational Neuroscience Center, West China Hospital, Sichuan University, Chengdu, 610041, China.
School of Pharmacy and Medical Sciences, Division of Health Sciences, University of South Australia, Adelaide, Australia.
出版信息
Stem Cell Res Ther. 2020 Apr 16;11(1):155. doi: 10.1186/s13287-020-01652-4.
BACKGROUND
The limited neuronal differentiation of the endogenous or grafted neural stem cells (NSCs) after brain injury hampers the clinic usage of NSCs. Panax notoginseng saponins (PNS) were extensively used for their clinical value, such as in controlling blood pressure, blood glucose, and inhibiting neuronal apoptosis and enhancing neuronal protection, but whether or not it exerts an effect in promoting neuronal differentiation of the endogenous NSCs is completely unclear and the potential underlying mechanism requires further exploration.
METHODS
Firstly, we determined whether PNS could successfully induce NSCs to differentiate to neurons under the serum condition. Mass spectrometry and quantitative polymerase chain reaction (Q-PCR) were then performed to screen the differentially expressed proteins (genes) between the PNS + serum and serum control group, upon which dihydropyrimidinase-like 2 (DPYSL2), a possible candidate, was then selected for the subsequent research. To further investigate the actual role of DPYSL2 in the NSC differentiation, DPYSL2-expressing lentivirus was employed to obtain DPYSL2 overexpression in NSCs. DPYSL2-knockout rats were constructed to study its effects on hippocampal neural stem cells. Immunofluorescent staining was performed to identify the differentiation direction of NSCs after 7 days from DPYSL2 transfection, as well as those from DPYSL2-knockout rats.
RESULTS
Seven differentially expressed protein spots were detected by PD Quest, and DPYSL2 was found as one of the key factors of NSC differentiation in a PNS-treated condition. The results of immunostaining further showed that mainly Tuj1 and GFAP-positive cells increased in the DPYSL2-overexpressed group, while both were depressed in the hippocampal NSCs in the DPYSL2-knockout rat.
CONCLUSIONS
The present study revealed that the differentiation direction of NSCs could be enhanced through PNS administration, and the DPYSL2 is a key regulator in promoting NSC differentiation. These results not only emphasized the effect of PNS but also indicated DPYSL2 could be a novel target to enhance the NSC differentiation in future clinical trials.
背景
脑损伤后内源性或移植的神经干细胞（NSCs）的神经元分化有限，限制了 NSCs 的临床应用。三七总皂苷（PNS）因其临床价值而被广泛应用，如控制血压、血糖、抑制神经元凋亡和增强神经元保护等，但它是否能促进内源性 NSCs 的神经元分化尚不清楚，其潜在的作用机制需要进一步探索。
方法
首先，我们确定 PNS 是否能在血清条件下成功诱导 NSCs 向神经元分化。然后进行质谱分析和定量聚合酶链反应（Q-PCR），筛选 PNS+血清组和血清对照组之间差异表达的蛋白质（基因），选择二氢嘧啶酶样 2（DPYSL2）作为可能的候选基因进行后续研究。为了进一步研究 DPYSL2 在 NSC 分化中的实际作用，我们使用表达 DPYSL2 的慢病毒获得 NSCs 中的 DPYSL2 过表达。构建 DPYSL2 敲除大鼠，研究其对海马神经干细胞的影响。转染 DPYSL2 7 天后进行免疫荧光染色，鉴定 NSCs 的分化方向，以及 DPYSL2 敲除大鼠的分化方向。
结果
PD Quest 检测到 7 个差异表达蛋白斑点，发现 DPYSL2 是 PNS 处理条件下 NSC 分化的关键因素之一。免疫染色结果进一步表明，DPYSL2 过表达组中 Tuj1 和 GFAP 阳性细胞增多，而 DPYSL2 敲除大鼠海马 NSCs 中两种细胞均减少。
结论
本研究表明，通过 PNS 给药可以增强 NSCs 的分化方向，DPYSL2 是促进 NSCs 分化的关键调节因子。这些结果不仅强调了 PNS 的作用，还表明 DPYSL2 可能成为未来临床试验中增强 NSCs 分化的新靶点。
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