Changes in Tear Protein Profile in Patients With Conjunctivochalasis
Changes in Tear Protein Profile in Patients With Conjunctivochalasis
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DOI：
10.1097/ico.0b013e3181dea7d7
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发表时间：
2011-01-01
期刊：
影响因子：
2.8
通讯作者：
Duran, Juan A.
中科院分区：
文献类型：
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作者：
Acera, Arantxa;Suarez, Tatiana;Duran, Juan A.
目的：比较正常志愿者和结膜松弛症(CCH)患者泪液的蛋白质谱，以确定在CCH病理过程中表达变化的蛋白质。方法：对8例正常人和6例CCH患者的泪液进行双向电泳分析。用等电聚焦法分离泪液总蛋白，用8%~16%十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法进行第二维分离。用Progensis SameSpot软件对凝胶图像进行分析。人工从凝胶中切下这些感兴趣的斑点，并通过基质辅助激光解吸/电离飞行时间(MALDI-TOF)鉴定相应的蛋白质。结果：在整个泪液蛋白质组中检测到约250个斑点蛋白。与对照组相比，CCH组有24个点显著上调。共鉴定出11个蛋白质斑点，其中包括S100家族的蛋白质(A8、A9、A4；分别上调2.44、1.71和2.82倍)，鸟苷三磷酸结合蛋白2(1.95倍)，L-乳酸脱氢酶A样6B(2.32倍)，脂肪酸结合蛋白(2.01倍)，角蛋白I型细胞骨架10(1.81倍)，谷胱甘肽转移酶P(2.27倍)，过氧化还蛋白-1，过氧化还蛋白-5(分别为1.79和1.92倍)，以及cullin-4B+甘油醛3-磷酸脱氢酶(1.96倍)。这是在CCH的眼泪中上调的。虽然其中一些，如S100A4、S100A8和过氧化还蛋白-5，是炎症和氧化过程的标志物，但监测它们在CCH中的水平可能有助于评估疾病的严重程度和进展。
Purpose: To compare the protein profiles of tears from normal volunteers and patients with conjunctivochalasis (CCH), with a view to identifying proteins whose expression is altered in this pathology.Methods: Tears from 8 normal subjects and 6 patients with CCH were analyzed by 2-dimensional electrophoresis. Total protein from tears was separated in the first dimension by isoelectric focusing, and the second dimension was carried out using 8%-16% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gel images were analyzed using Progenesis SameSpot software. Those spots of interest were manually cut out from the gels, and the corresponding proteins were identified by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF). Expression levels of proteins that had been found to be significantly altered were further verified by Western blot.Results: Approximately 250 spot proteins were detected in the whole tear proteome. Twenty-four spots were significantly upregulated in CCH compared with that in controls. Eleven protein spots were identified, which included proteins belonging to the S100 family (A8, A9, A4; 2.44, 1.71, and 2.82 fold upregulation, respectively), guanosine triphosphate-binding protein 2 (1.95 fold), L-lactate dehydrogenase A-like 6B (2.32 fold), fatty acid-binding protein (2.01 fold), keratin type I cytoskeletal 10 (1.81 fold), glutathione Stransferase P (2.27 fold), peroxiredoxin-1, peroxiredoxin-5 (1.79- and 1.92 fold, respectively), and cullin-4B+ glyceraldehyde 3-phosphate dehydrogenase (1.96 fold).Conclusions: We have identified a group of proteins, which is upregulated in CCH tears. Although some of them, such as S100A4, S100A8, and peroxiredoxin-5, are markers of inflammation and oxidative processes, monitoring their levels in CCH might be useful for assessing the severity and progression of the disease.