Effect of downregulated citrate synthase on oxidative phosphorylation signaling pathway in HEI-OC1 cells.

IF 1.6 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Proteome Science Pub Date : 2022-09-07 DOI:10.1186/s12953-022-00196-0

Xiaowen Xu, Yue Liu, Jun Luan, Rongrong Liu, Yan Wang, Yingying Liu, Ang Xu, Bingxin Zhou, Fengchan Han, Wenjing Shang

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引用次数: 1

Abstract

Background: Citrate Synthase (Cs) gene mutation (locus ahL4) has been found to play an important role in progressive hearing loss of A/J mice. HEI-OC1 cells have been widely used as an in vitro system to study cellular and molecular mechanisms related to hearing lose. We previously reported the increased apoptosis and the accumulation of reactive oxygen species in shRNACs-1429 cells, a Cs low-expressed cell model from HEI-OCI. The details of the mechanism of ROS production and apoptosis mediated by the abnormal expression of Cs needed to research furtherly.

Methods: iTRAQ proteomics was utilized to detect the differentially expressed proteins (DEPs) caused by low expression of Cs. The GO and KEGG pathways analysis were performed for annotation of the differentially expressed proteins. Protein-protein interaction network was constructed by STRING online database. Immunoblotting was utilized to confirm the protein levels of the the differentially expressed proteins.

Results: The differentially expressed proteins were significantly enriched in various signaling pathways mainly related to mitochondrial dysfunction diseases including Parkinson's disease, Alzheimer's disease, Huntington's disease, et al. Most noteworthy, the oxidative phosphorylation pathway was most significantly suppressed in the shRNACs-1429 cells,, in which a total of 10 differentially expressed proteins were enriched and were all downregulated by the abnormal expression of Cs. The downregulations of Ndufb5, Ndufv1 and Uqcrb were confirmed by immunoblotting. Meanwhile, the ATP levels of shRNACs-1429 cells were also reduced.

Conclusions: These results suggest that low level expression of Cs induces the inhibition of oxidative phosphorylation pathway, which is responsible for the high level production of reactive oxygen species and low level of ATP, leading to the apoptosis of cochlear cells. This study may provide new theories for understanding and therapy of progressive hearing loss.

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下调柠檬酸合酶对HEI-OC1细胞氧化磷酸化信号通路的影响。

背景:柠檬酸合酶(Citrate Synthase, Cs)基因突变位点ahL4在A/J小鼠进行性听力损失中起重要作用。HEI-OC1细胞作为一种体外系统被广泛应用于研究听力损失相关的细胞和分子机制。我们之前报道了HEI-OCI的Cs低表达细胞模型shRNACs-1429细胞凋亡增加和活性氧积累。Cs异常表达介导ROS产生和凋亡的具体机制有待进一步研究。方法:采用iTRAQ蛋白组学方法检测Cs低表达引起的差异表达蛋白(DEPs)。对差异表达蛋白进行GO和KEGG通路分析。利用STRING在线数据库构建蛋白-蛋白相互作用网络。采用免疫印迹法确定差异表达蛋白的蛋白水平。结果:差异表达蛋白在主要与帕金森病、阿尔茨海默病、亨廷顿病等线粒体功能障碍疾病相关的多种信号通路中显著富集。最值得注意的是，氧化磷酸化途径在shRNACs-1429细胞中受到的抑制最为显著，在shRNACs-1429细胞中，共有10种差异表达蛋白富集，且均因Cs的异常表达而下调。免疫印迹法证实Ndufb5、Ndufv1和Uqcrb下调。同时，shRNACs-1429细胞的ATP水平也降低。结论:低水平表达Cs可诱导耳蜗细胞氧化磷酸化通路受到抑制，导致高水平活性氧生成和低水平ATP生成，从而导致耳蜗细胞凋亡。本研究为理解和治疗进行性听力损失提供了新的理论依据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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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.