Department of Ophthalmology, Shanghai Changzheng Hospital, Shanghai, Shanghai, China.
Proteome Research Cente, Shanghai Applied Protein Technology, Shanghai, Shanghai, China.
PeerJ. 2022 Jul 12;10:e13701. doi: 10.7717/peerj.13701. eCollection 2022.
Thyroid-associated ophthalmopathy (TAO) is a common orbital inflammatory disease, but the abnormal expression of proteins in tears of TAO patients has not been systematically studied. The purpose of this study is to compare and analyze the total tear protein profile of TAO patients and to provide protein cues for TAO pathogenesis.
Tear samples were isolated from 30 TAO patients with obvious ocular surface damage and 30 healthy control subjects. Tear samples from 30 individuals were mixed and divided into three sample pools. Easy nano-scale LC-MS/MS based on labeling-free quantitative technology was utilized to profile tear proteome.
Here, electrospray ionization mass spectra and SDS-PAGE results confirmed the good parallelisms among samples. A total of 313 proteins were obtained from six tear pools, among them, 103 differential abundance proteins (DAPs) were identified, including 99 up-regulated DAPs (including APOA1, HV103, IGH, and Transferrin variant) and four down-regulated DAPs (including FABA, VCC1, NUCB2, and E-cadherin) in the TAO group compared with the control group. GO analysis showed that up-regulated DAPs were mainly enriched in lipid metabolism and platelet molecular function, and down-regulated DAPs were involved in binding, cell junction, and cellular process. KEGG results indicated that DAPs were involved in 117 kinds of signal transduction pathways, among which the immune-related pathway of complement and coagulation cascades had the greatest relevance.
In conclusion, label-free LC-MS/MS is an effective strategy for profiling tear proteins component. Our study provides proteins and pathways altered in TAO and provides protein cues for further study on the precise mechanism of TAO pathogenesis.
甲状腺相关眼病(TAO)是一种常见的眼眶炎性疾病,但 TAO 患者泪液中蛋白质的异常表达尚未得到系统研究。本研究旨在比较和分析 TAO 患者的总泪蛋白谱,为 TAO 发病机制提供蛋白线索。
从 30 例有明显眼表损伤的 TAO 患者和 30 例健康对照者中分离出泪液样本。将 30 例个体的泪液样本混合并分为三个样本池。利用基于无标记定量技术的 Easy nano-scale LC-MS/MS 对泪液蛋白质组进行分析。
在这里,电喷雾电离质谱和 SDS-PAGE 结果证实了样品之间的良好平行性。从六个泪液池中共获得 313 种蛋白质,其中 103 种差异丰度蛋白(DAPs)被鉴定出来,包括 TAO 组中 99 个上调的 DAPs(包括 APOA1、HV103、IGH 和转铁蛋白变体)和 4 个下调的 DAPs(包括 FABA、VCC1、NUCB2 和 E-钙黏蛋白)与对照组相比。GO 分析显示,上调的 DAPs 主要富集在脂质代谢和血小板分子功能中,而下调的 DAPs 则参与了结合、细胞连接和细胞过程。KEGG 结果表明,DAPs 参与了 117 种信号转导途径,其中补体和凝血级联的免疫相关途径相关性最大。
总之,无标记 LC-MS/MS 是一种有效的分析泪液蛋白成分的策略。本研究为 TAO 中改变的蛋白质和途径提供了线索,并为进一步研究 TAO 发病机制的精确机制提供了蛋白线索。