摘要
目的观察病理性近视(PM)患者房水标本中蛋白质表达谱的变化。方法横断面研究。2019年1~8月在天津医科大学眼科医院收集32例老年性白内障患者的房水样本进行质谱检测。其中,男性11例,女性21例;年龄58~76岁,平均年龄(68.41±6.09)岁。将合并PM的16例作为PM组,不合并近视的16例作为对照组。所有患者在白内障手术操作之前从手术眼收集100~150μl前房水。采用蛋白定量和非标记液相色谱串联质谱分析,得到差异表达蛋白。随机选取5个差异蛋白进行ELISA验证。然后用基因注释功能富集、京都基因和基因组百科全书通路富集等生物信息学分析方法分析差异表达蛋白的功能。结果两组房水标本中共鉴定出583个可定量蛋白质,其中101个蛋白存在差异表达,包括63个上调蛋白和38个下调蛋白。ELISA验证结果显示,5个差异表达蛋白在PM组和对照组之间的表达变化趋势均与非标记定量蛋白质组学分析结果相一致。这些差异表达蛋白的分类主要包括蛋白结合活性调节因子、防御/免疫蛋白、蛋白质修饰酶、代谢物间转换酶、细胞外基质蛋白等。生物信息学分析表明,PM与炎症和免疫相互作用以及细胞外基质的重塑密切相关。结论PM患者房水标本中蛋白质表达谱较对照组有明显变化,这些差异变化提示PM与炎症和免疫相互作用以及细胞外基质的重塑密切相关。
Objective To characterize proteomic profile in aqueous humor of patients with pathologic myopia(PM)using quantitative proteomic analysis,which may provide new clues to understand the mechanisms and possible treatments of PM.Methods A cross-sectional study.From January 2019 to August 2019,aqueous humor samples(32 cataract patients)were collected for quantitative proteomic analysis using liquid chromatography tandem mass spectrometry at Tianjin Medical University Eye Hospital.There were 11 males and 21 females.They were 58-76 years old with an average age of 68.41±6.09 years old.Sixteen patients with PM were regarded as PM group,16 patients without myopia were regarded as the control group.The aqueous humor samples(100-150μl)were collected from all patients before cataract surgery.Using protein quantification and non-labeled liquid chromatography tandem mass spectrometry analysis,differentially expressed proteins were obtained.Five different proteins were randomly selected for ELISA verification.The differentially expressed proteins were further analyzed by gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes,which were validated using ELISA in the other twenty samples of each group.Results A total of 583 proteins were identified and 101 proteins were found to be differentially expressed,including 63 up-regulated proteins and 38 down-regulated proteins.ELISA verification results showed that the expression trend of the 5 differentially expressed proteins between the PM group and the control group was consistent with the results of Label-free quantitative proteomics analysis.The main classifications of these differentially expressed proteins were protein-binding activity modulator,defense/immunity protein,protein modifying enzyme,metabolite interconversion enzyme,extracellular matrix protein,transfer/carrier protein and so on.The bioinformatics analysis suggested that PM was closely associated with inflammation and immune interactions,and remodeling of extracellular matrix.Conclusions Compared with the control group,the protein expression profile of PM patients'aqueous humor specimens has obvious changes.These differences indicate that PM is closely related to inflammation and immune interaction and extracellular matrix remodeling.
作者
薛敏
柯屹峰
任新军
刘巨平
范小娥
李筱荣
Xue Min;Ke Yifeng;Ren Xinjun;Liu Juping;Fan Xiao’e;Li Xiaorong(Tianjin Key Laboratory of Retinal Functions and Diseases,Tianjin International Joint Research and Development Centre of Ophthalmology and Vision Science,Eye Institute and School of Optometry,Tianjin Medical University Eye Hospital,Tianjin 300384,China;Department of Ophthalmology of Anhui NO.2 Provincial People’s Hospital,Hefei 230041,China)
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2020年第12期936-942,共7页
Chinese Journal of Ocular Fundus Diseases
基金
天津医科大学眼科医院高水平创新型人才培养基金杰出学者项目(YDYYRCXM-A2018-01)。
关键词
近视
退行性
眼房水
蛋白质组学
Myopia,degenerative
Aqueous humor
Proteomics
