Subasi Sevgi, Altintas Ozgul, Kasap Murat, Guzel Nil, Akpinar Gurler, Karaman Suleyman
Department of Ophthalmology, Körfez State Hospital, Kocaeli, Turkey.
Department of Ophthalmology, Medical Faculty, Acıbadem University, Istanbul, Turkey.
Int Ophthalmol. 2018 Dec;38(6):2563-2574. doi: 10.1007/s10792-017-0770-5. Epub 2017 Nov 23.
In this study, by using a two-dimensional (2D) electrophoresis-based experimental approach, we aimed at understanding the nature of alkali injuries and the underlying mechanisms. A secondary aim was to compare the effects of cross-linking (CXL) and amnion membrane transplantation (AMT) on corneal protein compositions at the end of the early repair phase after injured with alkali.
The right corneas of 24 rabbits were injured with a 1 N solution of NaOH. Groups were formed based on the adjuvant therapies as (1) healthy group, (2) control group, (3) CXL group, (4) AMT group. In addition to the therapies, a conventional medical treatment was applied to all groups. Left eyes were used as within-subject healthy corneas (1). The corneas were excised at day 21, and a comparative proteomic analysis was performed using 2D gel electrophoresis coupled with MALDI-TOF/TOF.
2D gel electrophoresis revealed the presence seven protein spots whose abundance changed among the groups. Those proteins were SH3 domain-binding protein, plant homeodomain finger protein 23, S100 calcium binding protein A-11(S100 A11), keratin type 2 cytoskeletal 1 and 2, transketolase and glyceraldehyde 3-phosphate dehydrogenase. Ingenuity pathway analysis predicted that the observed changes may be linked to a central metabolic pathway, transforming growth factor beta 1. Canonical pathway analysis focused our attention to two different pathways, namely nicotinamide adenine dinucleotide repair pathway and non-oxidative branch of pentose phosphate pathway.
Our results shed some light onto the molecular mechanisms affected by alkali injury and adjuvant treatments. Further research is needed to propose medically significant target molecules that may be used for novel drug developments for alkali injury.
在本研究中，我们采用基于二维（2D）电泳的实验方法，旨在了解碱烧伤的本质及其潜在机制。第二个目的是比较交联（CXL）和羊膜移植（AMT）对碱烧伤后早期修复阶段末期角膜蛋白质组成的影响。
用1N的NaOH溶液损伤24只兔的右眼。根据辅助治疗方法将其分为（1）健康组、（2）对照组、（3）CXL组、（4）AMT组。除这些治疗外，所有组均采用常规药物治疗。左眼用作受试者自身的健康角膜（对照）。在第21天切除角膜，使用二维凝胶电泳结合基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF/TOF）进行比较蛋白质组学分析。
二维凝胶电泳显示有7个蛋白点在各组间丰度发生变化。这些蛋白质是SH3结构域结合蛋白、植物同源结构域指蛋白23、S100钙结合蛋白A-11（S100 A11）、角蛋白2型细胞骨架1和2、转酮醇酶和甘油醛-3-磷酸脱氢酶。通路分析预测，观察到的变化可能与一条中心代谢通路——转化生长因子β1有关。经典通路分析将我们的注意力集中在两条不同的通路上，即烟酰胺腺嘌呤二核苷酸修复通路和磷酸戊糖途径非氧化分支。
我们的结果揭示了受碱烧伤和辅助治疗影响的分子机制。需要进一步研究以提出具有医学意义的靶分子，可用于开发治疗碱烧伤的新型药物。
