原发性急性闭角型青光眼患者房水蛋白质组学分析

Proteomic Analysis of Aqueous Humor Proteins in Patients with Primary Acute Angle-Closure Glaucoma

目的:探究不同眼压的原发性急性闭角型青光眼(PAACG)患者房水蛋白的差异性表达,寻找青光眼性视神经损伤的可能机制,为青光眼性视神经保护的可能作用靶点提供实验依据。方法:病例对照研究。通过简单随机抽样选取2019年3月至2020年9月期间于吉林大学第二医院眼科中心入院治疗的PAACG患者共88例(88眼)。根据PAACG患者的眼压将88个选定的样本分为2组:A组36个样本,眼压≥50 mmHg(1 mmHg=0.133 kPa);B组52个样本,眼压≤21 mmHg。以上样本被分为2个队列:发现队列(A组26个样本;B组37个样本)和验证队列(A组10个样本;B组15个样本)。分别通过数据独立采集(DIA)方法和平行反应监测(PRM)方法分析房水蛋白。采用独立样本t检验分析蛋白表达差异。结果:本研究从发现队列的A、B组中63个房水样本中共检测出636个蛋白,去除默认蛋白后得到506个蛋白用于后续分析,其中51个蛋白在发现队列A、B组间差异有统计学意义(P=0.020)。在这51个差异蛋白中,A组存在17个蛋白表达上调,B组存在34个蛋白表达上调。抽取上述51个差异蛋白中APOA2、TIMP1、LRP2和VASN进行PRM验证,结果显示这4个差异蛋白在发现队列及验证队列中保持一致。结论:PAACG患者房水中差异蛋白的表达与炎症反应以及小梁网细胞外基质重塑、神经损伤相关,可能是导致青光眼患者眼压升高以及视神经损伤的重要原因。

Objective:Explore the differential expression of aqueous protein in patients with primary acute angle-closure glaucoma(PAACG)with different intraocular pressure,find the possible mechanism of glaucomatous optic nerve damage,and provide experimental basis for possible targets for glaucomatous optic nerve protection.Methods:In this case-control study,a total of 88 patients(88 eyes)with primary acute angle-closure glaucoma admitted to the Ophthalmology Center of the Second Hospital of Jilin University from March 2019 to September 2020 were randomly collected.Eighty-eight selected samples were divided into two groups according to the intraocular pressure(IOP)of patients with PAACG:group A(36 samples,IOP 50 mmHg,1 mmHg=0.133 kPa)and group B(52 samples,IOP 21 mmHg).The above samples were divided into two cohorts:discovery cohort(group A,26 samples;group B,37 samples)and validation cohort(group A,10 samples;group B,15 samples).Their aqueous humor proteomes were analyzed by the data-independent acquisition method and by the parallel reaction monitor method(PRM),respectively.Indepenednt t-test was used to compare protein expression.Results:In this study,a total of 636 proteins were detected from 63 aqueous humor samples in groups A and B of the discovery queue,and 506 proteins were obtained for subsequent analysis after the removal of the default protein.Among them,51 proteins were differentially expressed between the two groups(P=0.020).Moreover,17 proteins of these 51 differential proteins were up-regulated in group A,and 34 proteins were up-regulated in group B.Besides,APOA2,TIMP1,LRP2,and VASN were randomly selected for PRM validation,and the study results demonstrated that the results in the discovery queue were consistent with those in the validation queue.Conclusions:Our study results revealed that the expression of differentially expressed proteins was associated with inflammatory response,as well as extracellular matrix remodeling in the trabecular meshwork and nerve injury.The above factors would be an essential reason for increased intraocular pressure and optic nerve injury in glaucoma.