Proteomic Analysis of Protective Effects of Dl-3-n-Butylphthalide against mpp + -Induced Toxicity via downregulating P53 pathway in N2A Cells.

IF 2.1 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS Proteome Science Pub Date : 2023-01-03 DOI:10.1186/s12953-022-00199-x

Yuan Zhao, Jian Zhang, Yidan Zhang, Shuyue Li, Ya Gao, Cui Chang, Xiang Liu, Lei Xu, Guofeng Yang

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引用次数: 3

Abstract

Background: Dl-3-n-butylphthalide (NBP) is an important medial therapy for acute ischemic stroke in China. Recent studied have revealed that NBP not only rescued the loss of dopaminergic neurons in cellular and animal models of Parkinson's disease (PD), but also could improve motor symptoms in PD patients. However, the protective mechanism is not fully understood. P53 is a multifunctional protein implicated in numerous cellular processes, including apoptosis, DNA repair, mitochondrial functions, redox homeostasis, autophagy and protein aggregations. In PD, p53 integrated with various neurodegeneration-related signals inducing neuronal loss, indicating the suppression of P53 might be a promising target for PD treatment. Therefore, the purpose of the current study was to systemically screen new therapeutic targets of NBP in PD.

Method: In our study, we constructed mpp + induced N2A cells to investigate the benefit effect of NBP in PD. MTT assay was performed to evaluate the cell viability; TMT-based LC-MS/MS was applied to determine the different expressed proteins (DEPs) of NBP pretreatment; online bioinformatics databases such as DAVID, STRING, and KEGG was used to construe the proteomic data. After further analyzed and visualized the protein-protein interactions (PPI) by Cytoscape, DEPs were verified by western blot.

Result: A total of 5828 proteins were quantified in the comparative proteomics experiments and 417 proteins were considered as DEPs (fold change > 1.5 and p < 0.05). Among the 417 DEPs, 140 were upregulated and 277 were downregulated in mpp + -induced N2A cells with NBP pretreatment. KEGG pathway analysis indicated that lysosome, phagosome, apoptosis, endocytosis and ferroptosis are the mainly enriched pathways. By using MCL clustering in PPI analysis, 48 clusters were generated and the subsequent KEGG analysis of the top 3 clusters revealed that P53 signaling pathway was recognized as the dominant pathway for NBP treatment.

Conclusion: NBP significantly relived mpp + -induced cell toxicity. The neuroprotective role of NBP was implicated with P53 signaling pathway in some extent. These findings will reinforce the understanding of the mechanism of NBP in PD and identify novel therapeutic targets.

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dl -3-n-丁苯酞通过下调P53通路对N2A细胞mpp +诱导毒性的保护作用的蛋白质组学分析。

背景:dl -3-正丁苯酞(NBP)在中国是治疗急性缺血性脑卒中的重要药物。最近的研究表明，NBP不仅可以挽救帕金森病(PD)细胞和动物模型中多巴胺能神经元的损失，而且可以改善PD患者的运动症状。然而，保护机制尚不完全清楚。P53是一种涉及多种细胞过程的多功能蛋白，包括细胞凋亡、DNA修复、线粒体功能、氧化还原稳态、自噬和蛋白质聚集。在PD中，p53与多种神经退行性相关信号结合，诱导神经元丢失，表明抑制p53可能是PD治疗的一个有希望的靶点。因此，本研究的目的是系统筛选NBP治疗PD的新靶点。方法:构建mpp +诱导的N2A细胞，研究NBP对PD的益处作用。MTT法测定细胞活力;采用基于tmt的LC-MS/MS检测NBP预处理的不同表达蛋白(DEPs);利用DAVID、STRING和KEGG等在线生物信息学数据库构建蛋白质组学数据。通过Cytoscape进一步分析和可视化蛋白相互作用(PPI)后，用western blot验证DEPs。结果:在比较蛋白质组学实验中，共有5828个蛋白被量化，其中417个蛋白被认为是DEPs (fold change > 1.5和p)。结论:NBP可显著缓解mpp +诱导的细胞毒性。NBP的神经保护作用可能与P53信号通路有关。这些发现将加强对NBP在PD中的作用机制的理解，并确定新的治疗靶点。

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来源期刊

Proteome Science 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

审稿时长

4.5 months

期刊介绍： Proteome Science is an open access journal publishing research in the area of systems studies. Proteome Science considers manuscripts based on all aspects of functional and structural proteomics, genomics, metabolomics, systems analysis and metabiome analysis. It encourages the submissions of studies that use large-scale or systems analysis of biomolecules in a cellular, organismal and/or environmental context. Studies that describe novel biological or clinical insights as well as methods-focused studies that describe novel methods for the large-scale study of any and all biomolecules in cells and tissues, such as mass spectrometry, protein and nucleic acid microarrays, genomics, next-generation sequencing and computational algorithms and methods are all within the scope of Proteome Science, as are electron topography, structural methods, proteogenomics, chemical proteomics, stem cell proteomics, organelle proteomics, plant and microbial proteomics. In spite of its name, Proteome Science considers all aspects of large-scale and systems studies because ultimately any mechanism that results in genomic and metabolomic changes will affect or be affected by the proteome. To reflect this intrinsic relationship of biological systems, Proteome Science will consider all such articles.