Tian Han, Wang Leilei, Cai Ruiqi, Zheng Ling, Guo Lin
College of Life Sciences, Wuhan University, Wuhan, China.
PLoS One. 2014 Dec 30;9(12):e116453. doi: 10.1371/journal.pone.0116453. eCollection 2014.
Retinal ischemia is a common feature associated with several ocular diseases, including diabetic retinopathy. In this study, we investigated the effect of a retinal ischemia and reperfusion (I/R) injury on protein levels via a quantitative shotgun strategy using stable isotope dimethyl labeling combined with LC-MS/MS analysis. Based on the relative quantitation data of 1088 proteins, 234 proteins showed a greater than 1.5-fold change following I/R injury, 194 of which were up-regulated and 40 were down-regulated. Gene ontology analysis revealed that after I/R injury, there was an increase in the metabolic-process related proteins but a decline in cell communication, system process and transport-related proteins. A ribosome protein network and a secreted protein network consisting of many protease inhibitors were identified among the up-regulated proteins, despite a suppression of the mammalian target of rapamycin (mTOR) pathway following the I/R injury. A synaptic-related protein network was found to be significantly down-regulated, implicating a functional reduction of neurons following a retinal I/R injury. Our results provide new systems-biology clues for the study of retinal ischemia.
视网膜缺血是包括糖尿病视网膜病变在内的多种眼部疾病的一个常见特征。在本研究中，我们通过使用稳定同位素二甲基标记结合液相色谱 - 串联质谱分析的定量鸟枪法策略，研究了视网膜缺血再灌注（I/R）损伤对蛋白质水平的影响。基于1088种蛋白质的相对定量数据，234种蛋白质在I/R损伤后显示出大于1.5倍的变化，其中194种上调，40种下调。基因本体分析显示，I/R损伤后，与代谢过程相关的蛋白质增加，但细胞通讯、系统过程和运输相关的蛋白质减少。尽管I/R损伤后雷帕霉素靶蛋白（mTOR）通路受到抑制，但在上调的蛋白质中鉴定出了一个核糖体蛋白网络和一个由许多蛋白酶抑制剂组成的分泌蛋白网络。发现一个与突触相关的蛋白质网络显著下调，这表明视网膜I/R损伤后神经元功能降低。我们的结果为视网膜缺血研究提供了新的系统生物学线索。
