Comparative proteome analysis of form-deprivation myopia in sclera with iTRAQ-based quantitative proteomics.

IF 1.8 3区医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Vision Pub Date : 2021-09-01 eCollection Date: 2021-01-01

Ying Yuan, Chengcheng Zhu, Mingming Liu, Bilian Ke

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Abstract

Objective: Scleral remodeling plays a key role in axial elongation in myopia. The aim of the present study was to identify the proteomics changes and specific signaling networks to gain insight into the molecular basis of scleral remodeling in myopic eyes.

Methods: Guinea pig form-deprivation myopia was induced with a translucent diffuser on a random eye for 4 weeks, while the other eye served as the contralateral control group. The axial length and refraction were measured at the beginning and end of the treatment. The proteins were extracted from the sclerae of each group and prepared for quantitative isobaric tags for relative and absolute quantification (iTRAQ) labeling combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The coexpression networks and protein functions were analyzed using Gene Ontology (GO) and Ingenuity Pathway Analysis (IPA). Quantitative real-time PCR (qRT-PCR) and western blotting were performed to confirm the authenticity and accuracy of the iTRAQ results.

Results: After 4 weeks, the form-deprivation eyes developed significant degrees of myopia, and the axial length increased statistically significantly (p<0.05). A total of 2,579 unique proteins with <1% false discovery rate (FDR) were identified. Furthermore, 56 proteins were found to be upregulated, and 84 proteins were found to be downregulated, with a threshold of a 1.2-fold change and p<0.05 in the myopia group, when compared to the control group. Further bioinformatics analysis indicated that 44 of 140 differentially expressed proteins were involved in cellular movement and cellular assembly and organization. The qRT-PCR or western blotting results confirmed that myosin IIB, ACTIN3, and cellular cytoskeletons were downregulated, while RhoA and RAP1A were upregulated in the sclera in myopic eyes. These results were consistent with the proteomics results.

Conclusions: Proteomics and bioinformatics results can be helpful for identifying proteins and providing new insights for better understanding of the molecular mechanism underlying scleral remodeling. These results revealed that the proteins associated with cellular movement and cellular assembly and organization are altered during the development of myopia. Furthermore, RhoA plays a key role in the pathways involved in cellular movement and cellular assembly and organization.

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基于itraq的巩膜形态剥夺性近视定量蛋白质组学比较分析。

目的:巩膜重塑在近视眼轴伸长中起关键作用。本研究的目的是鉴定蛋白质组学变化和特定的信号网络，以深入了解近视眼睛巩膜重塑的分子基础。方法:随机取一只眼用半透明扩散器诱导豚鼠形态剥夺性近视4周，另一只眼作为对侧对照组。在治疗开始和结束时测量眼轴长和折射。从各组细胞的硬膜中提取蛋白质，制备定量等压标签，用于相对定量和绝对定量(iTRAQ)标记，并结合液相色谱-串联质谱(LC-MS/MS)分析。利用基因本体(Gene Ontology, GO)和匠心途径分析(Ingenuity Pathway Analysis, IPA)对共表达网络和蛋白功能进行分析。采用实时荧光定量PCR (qRT-PCR)和western blotting验证iTRAQ结果的真实性和准确性。结果:4周后，形态剥夺眼发生了明显的近视程度，眼轴长度明显增加(p)结论:蛋白质组学和生物信息学结果有助于鉴定蛋白质，为更好地理解巩膜重塑的分子机制提供新的见解。这些结果表明，在近视的发展过程中，与细胞运动和细胞组装和组织相关的蛋白质发生了改变。此外，RhoA在参与细胞运动和细胞组装和组织的途径中起着关键作用。

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来源期刊

Molecular Vision 生物-生化与分子生物学

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： Molecular Vision is a peer-reviewed journal dedicated to the dissemination of research results in molecular biology, cell biology, and the genetics of the visual system (ocular and cortical). Molecular Vision publishes articles presenting original research that has not previously been published and comprehensive articles reviewing the current status of a particular field or topic. Submissions to Molecular Vision are subjected to rigorous peer review. Molecular Vision does NOT publish preprints. For authors, Molecular Vision provides a rapid means of communicating important results. Access to Molecular Vision is free and unrestricted, allowing the widest possible audience for your article. Digital publishing allows you to use color images freely (and without fees). Additionally, you may publish animations, sounds, or other supplementary information that clarifies or supports your article. Each of the authors of an article may also list an electronic mail address (which will be updated upon request) to give interested readers easy access to authors.