原发性开角型青光眼房水差异表达蛋白分析

Analysis of different proteins in primary open angle glaucoma based on aqueous proteome

目的分析原发性开角型青光眼(POAG)患者房水的蛋白质表达变化,探寻与疾病发生相关的生物学标志及潜在治疗靶点.方法采用回顾性病例-对照研究设计.纳入2016年10月至2017年12月由天津医科大学眼科医院收治的行白内障手术的年龄相关性白内障患者10例10眼作为年龄相关性白内障组,行青光眼联合白内障手术的POAG合并白内障患者10例10眼作为POAG合并白内障组.术中借助手术通道用1号针头接1ml针筒进入前房中部吸取约100μl房水.通过非标记定量蛋白质组学质谱分析技术分析房水中提取的蛋白,采用Maxquant significancesA的方法进行差异显著性检验,以P<0.05、差异倍数>2的标准筛选得到2个组患者的差异蛋白,并将生物大数据通过GO功能、KEGG显著性富集分析对差异蛋白的功能及调控的信号通路加以注解.结果本次蛋白组学分析共检测到97个差异蛋白,其中包括48个上调蛋白和49个下调蛋白.GO分析显著差异蛋白功能涉及诸多方面,其中与炎症反应有关的有脂多糖结合蛋白(LBP)、CD163、C-反应蛋白(CRP)和膜联蛋白A1(ANXA1);与氧化还原有关的蛋白有谷胱甘肽S转移酶P(GSTP1)和硫氧还原蛋白(TXN);与细胞黏附运动有关:软骨寡聚基质蛋白(COMP)、桥粒胶蛋白(DSC2)和层连蛋白(LAMB2);与纤维化有关的有原胶原蛋白(PLOD1)和固生蛋白(TNC);与神经生长有关:颤蛋白(RELN)、脑信号蛋白(SEMA3F)和轴突导向因子(SEMA4B);与代谢相关的蛋白有丙酮酸激酶(PKM)和羧肽酶N亚型2(CPN2)等.KEGG分析表明NrF2/ERK信号通路、TGF-β/NF-κB信号通路表达在2个组细胞间存在差异.结论POAG患者房水中GSTP1、TXN表达显著降低,可能通过调控NrF2/ERK1/2及TGF-β/NF-κB信号通路,调节细胞黏附性和活性以及细胞外基质表达来参与疾病的发展.GSTP1、TXN可能成为潜在生物学标志及治疗靶点.

Objective To analyze the changes of protein expression in aqueous humor of patients with primary open angle glaucoma (POAG), and to explore the biological markers and potential therapeutic targets associated with disease occurrence. Methods A retrospective case-control study was adopted.Ten patients with age-related cataract and 10 patients with POAG combined with cataract.were collected at the Tianjin Medical University Eye Hospital from October 2016 to December 2017.In the process of surgery, 100 μl of aqueous humor were collected with 1 ml syringe and No.1 needle through the surgical access.Those proteins extracted from aqueous humor were analyzed by quantitative proteomic mass spectrometry.The differential significance test was performed by Maxquant significances A. The differential proteins of the two groups were screened and determined with the conditions of P<0.05 and difference multiple>2.The biological big data was annotated by the function of GO and KEGG.The study was approved by the Ethics Committee of the Tianjin Medical University Eye Hospital (No.2013KY[L]-10). Patients and their guardians all signed the informed consent. Results Ninty-seven differential proteins were detected in this proteomic analysis, including 48 up-regulated proteins and 49 down-regulated proteins.GO analysis significantly differs in protein function involved in many aspects, some different proteins including lipopolysaccharide-binding protein (LBP), cluster of Differentiation 163(CD163), C-reactive protein (CRP), annexin A1 (ANXA1) were involved in inflammation;redox-related proteins were glutathione S-transferase P (GSTP1), thioredoxin (TXN), some different proteins related with cell adhesion movement were cartilage oligomeric matrix protein (COMP), desmocollin-2 (DSC2) and laminin subunit beta-2 (LAMB2);procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 (PLOD1), solid protein (tenascin, TNC) are associated with fibrosis;some different proteins related to nerve growth were reelin (RELN), semaphorin-3F(SEMA3F), semaphorin-4B(SEMA4B). Metabolism-related proteins included pyruvate kinase (PKM), carb oxypeptidase N subunit 2 (CPN2) and so on.KEGG analysis indicated that the expression of NrF2/ERK signaling pathway and TGF-β/NF-κB signaling pathway were different between the two groups. Conclusions The expression of GSTP1 and TXN in the aqueous humor of POAG is significantly decreased, which may regulate cell adhesion and activity and expression of extracellular matrix by regulating NrF2/ERK1/2 and TGF-β/NF-κB signaling pathways.GSTP1 and TXN may serve as a potential biomarker and therapeutic target of POAG.