摘要目的 分析急性原发性闭角型青光眼(APACG)急性发作期房水蛋白表达差异.方法 病例对照研究.收集2016年10月至2017年6月在天津医科大学眼科医院拟手术治疗的APACG急性发作期合并白内障患者(APACG合并白内障组)与白内障患者(白内障组)各10例(10只眼).征得患者知情同意后,术中借助手术通道用1号针头接1 ml针筒进入前房中部吸取50μl房水,注入无菌收集管中,-80℃保存.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 significances A的方法进行差异显著性检验,以P<0.05、差异倍数>2的标准筛选得到两组患者的差异蛋白,并将生物学大数据通过基因本体(GO)功能分析、京都基因与基因组百科全书(KEGG)信号通路显著性富集分析对两组患者房水差异蛋白的功能及调控的信号通路加以注解.结果 APACG合并白内障组男性3例,女性7例,年龄(68±6)岁;白内障组男性2例,女性8例,年龄(71±8)岁,两组性别分布及年龄差异均无统计学意义(均P>0.05).蛋白质组学分析显示与白内障组比,APACG合并白内障组房水中共检测到91个差异蛋白,包括50个上调蛋白(如膜联蛋白A1、波形蛋白、S100钙结合蛋白A8、白细胞介素6、C反应蛋白、层黏连蛋白 β2等)和41个下调蛋白(如角蛋白85、γ-晶状体蛋白D、突触结合蛋白5、脑信号蛋白4B、母系蛋白2、组织蛋白酶O、钙黏蛋白4、脑信号蛋白3B、血小板衍生生长因子D、转化生长因子β 等).GO分析显示两组房水中差异蛋白功能涉及诸多方面,其中膜联蛋白A1、CD163、S100钙结合蛋白A8、C反应蛋白、白细胞介素6参与炎性反应,钙黏蛋白4、层黏连蛋白 β2介导细胞黏附,母系蛋白2、波形蛋白、层黏连蛋白 β2等参与组织纤维化.KEGG显著性富集分析显示两组房水中差异蛋白主要参与磷脂酰肌醇3-激酶-蛋白激酶B信号通路、转化生长因子β相关信号通路、丝裂原活化蛋白激酶信号通路、Toll样受体信号通路、核因子-κB信号通路、黏附斑激酶信号通路和细胞外基质受体相互作用通路.结论 APACG急性发作期房水中膜联蛋白A1的表达显著上调,转化生长因子β、钙黏蛋白4和母系蛋白2等蛋白因子下调,房水中蛋白的改变可能与APACG急性发作有关.

abstractsObjective To analyze the difference among expression of aqueous humor proteins in acute primary angle-closure glaucoma (APACG). Methods Case-control study. The patients with APACG combined cataract (APACG with cataract group) and patients with cataract (cataract group), who had undertaken surgical treatment at the Tianjin Medical University Eye Hospital from October 2016 to June&nbsp;2017 were collected. Upon receipt of patient's consent, 50 μl of aqueous humor were collected with 1 ml syringe and No. 1 needle through the surgical access during the surgery, and then injected into a sterile collection tube to be stored at - 80 ℃ . Those proteins extracted from aqueous humor were analyzed by quantitative proteomic mass spectrometry. The differential significance test was performed by Maxquant significances A approach. The differential proteins of the two groups were screened and determined with the conditions of P<0.05 and difference multiple>2. The functions and signal pathway of differential proteins in aqueous humor were annotated in biological big data, on the basis gene ontology (GO) and the Kyoto gene and genomic encyclopedia (KEGG) analyses. Results There were 3 males and 7 females with an average age of (68±6) years in the APACG group. The cataract group included 2 males and 8 females with an average age of (71±8) years. There were no statistical differences in gender ratio and age between the two groups (both P>0.05). A total of 91 differential proteins were detected in this experiment, including 50 up-regulated proteins (annexinA1, vimentin, S100 calcium binding protein A8, interleukin 6, C reactive protein, laminin β2, etc.) and 41 down-regulated (keratin 85, γ-crystallin D, syntaxin-binding protein 5, semaphoring 4B, matrilin 2, cathepsin O, cadherin 4, semaphoring 3B, platelet-derived growth factor D, transforming growth factor β, etc.). On one hand, the functions of differential proteins involved in many aspects. AnnexinA1, CD163, S100 calcium-binding protein A8, C reactive protein, interleukin 6 are involved in the inflammatory reaction, cadherin 4 and laminin β2 regulate cell adhesion, matrilin 2, vimentin and laminin β2 participate in tissue fibrosis; on the other hand, KEGG analysis showed that the differential proteins participate diverse signaling pathways such as phosphatidylinositol-3-kinase-protein kinase B signaling pathway, transformation growth factor β signaling pathway, mitogen activated protein kinase signaling pathway, Toll-like receptor signaling pathway, the nuclear factor κ-light chain enhancer of the activated B cells signaling pathway, focal adhension and extracellular matrix receptor interaction pathway and so on. Conclusions The expression of annexin A1 is significantly up-regulated in the aqueous humor in APACG, while some other factors such as transformation growth factor β, cadherin-4, and matrilin 2 are down-regulated. The change of proteins in aqueous humor is related with the outbreak of APACG.