Wang L M, Dong L J, Liu X, Huang L Y, Liu W, Lyu Y J, Li X R, Liu A H
Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin Key Laboratory of Retinal Functions and Diseases, Tianjin 300384, China.
Zhonghua Yan Ke Za Zhi. 2019 Sep 11;55(9):687-694. doi: 10.3760/cma.j.issn.0412-4081.2019.09.011.
To analyze the difference among expression of aqueous humor proteins in acute primary angle-closure glaucoma (APACG). 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 June2017 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 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. 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 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. 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. .
分析急性原发性闭角型青光眼(APACG)房水蛋白质表达的差异。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接受手术治疗的APACG合并白内障患者(APACG合并白内障组)和白内障患者(白内障组)。获得患者同意后,在手术过程中用1 ml注射器和1号针头经手术切口收集50 μl房水,然后注入无菌收集管中,于-80℃保存。采用定量蛋白质组质谱分析法对从房水中提取的蛋白质进行分析。采用Maxquant significances A方法进行差异显著性检验。以P<0.05和差异倍数>2为条件筛选并确定两组的差异蛋白质。基于基因本体(GO)和京都基因与基因组百科全书(KEGG)分析,在生物大数据中注释房水差异蛋白质的功能和信号通路。APACG组有男性3例,女性7例,平均年龄(68±6)岁。白内障组有男性2例,女性8例,平均年龄(71±8)岁。两组性别比和年龄差异均无统计学意义(均P>0.05)。本实验共检测到91种差异蛋白质,其中上调蛋白质50种(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、层粘连蛋白β2等),下调蛋白质41种(角蛋白85、γ-晶状体蛋白D、Syntaxin结合蛋白5、信号素4B、基质金属蛋白酶2、组织蛋白酶O、钙黏蛋白4、信号素3B、血小板衍生生长因子D、转化生长因子β等)。一方面,差异蛋白质的功能涉及多个方面。膜联蛋白A1、CD163、S100钙结合蛋白A8、C反应蛋白、白细胞介素6参与炎症反应,钙黏蛋白4和层粘连蛋白β2调节细胞黏附,基质金属蛋白酶2、波形蛋白和层粘连蛋白β2参与组织纤维化;另一方面,KEGG分析表明差异蛋白质参与磷脂酰肌醇-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、活化B细胞核因子κ轻链增强子信号通路、黏着斑和细胞外基质受体相互作用通路等多种信号通路。APACG患者房水中膜联蛋白A1表达显著上调,而转化生长因子β、钙黏蛋白-4和基质金属蛋白酶2等其他一些因子表达下调。房水蛋白质变化与APACG的发病有关。