Zhou Yun-Yun, Chen Chang-Zheng, Su Yu, Li Lu, Yi Zuo-Hui-Zi, Qi Hang, Weng Ming, Xing Yi-Qiao
Department of Ophthalmology, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, Guangdong Province, China.
Int J Ophthalmol. 2014 Feb 18;7(1):8-13. doi: 10.3980/j.issn.2222-3959.2014.01.02. eCollection 2014.
To investigate the protective mechanism of Gingko Biloba extract (EGb761) on the ability of retinal pigment epithelial (RPE) cells to resist light-induced damage in a comparative proteomics study.
Human RPE cells (ARPE-19) were randomly distributed to one of three groups: normal control (NC group) and light-damaged model without or with EGb761 group (M and ME groups, respectively). The light-damaged model was formed by exposing to white light (2 200±300)lx for 6h. The RPE cells in ME group were conducted with EGb 761 (100µg/mL) before light exposure. The soluble cellular proteins extracting from each groups were separated by two-dimensional electrophoresis and stained by silver staining. Different proteins in the profiles of the gels were analyzed by Image Master Software. Two-fold expressing protein spots were identified by Matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) mass spectrometry.
NC, M and ME groups displayed 1 892±71, 2 145±23 and 2 216±85 protein spots, respectively. We identified 33 proteins with different expression levels between the NC and M groups, 25 proteins between the M and ME groups, and 11 proteins between the NC and ME groups. MALDI-TOF/TOF mass spectrometry successfully identified 16 proteins, including metabolic enzymes, cytoskeletal proteins, anti-oxidation proteins, and others.
Differences in some important proteins, such as cathepsin B, heat shock protein, and cytochrome c reductase, indicated that multiple pathways may be induced in light-damaged RPE cells and the protective effect of EGb761.
在一项比较蛋白质组学研究中，探讨银杏叶提取物（EGb761）对视网膜色素上皮（RPE）细胞抵抗光诱导损伤能力的保护机制。
将人RPE细胞（ARPE-19）随机分为三组之一：正常对照组（NC组）以及无光损伤模型组和有EGb761的光损伤模型组（分别为M组和ME组）。通过暴露于（2200±300）lx的白光6小时形成光损伤模型。ME组的RPE细胞在光照前用EGb 761（100μg/mL）处理。从每组中提取的可溶性细胞蛋白通过二维电泳分离并用银染法染色。凝胶图谱中的不同蛋白质通过Image Master软件进行分析。通过基质辅助激光解吸/电离串联飞行时间（MALDI-TOF/TOF）质谱鉴定两倍表达差异的蛋白质点。
NC、M和ME组分别显示1892±71、2145±23和2216±85个蛋白质点。我们鉴定出NC组和M组之间有33种表达水平不同的蛋白质，M组和ME组之间有25种，NC组和ME组之间有11种。MALDI-TOF/TOF质谱成功鉴定出16种蛋白质，包括代谢酶、细胞骨架蛋白、抗氧化蛋白等。
组织蛋白酶B、热休克蛋白和细胞色素c还原酶等一些重要蛋白质的差异表明，光损伤的RPE细胞可能诱导多种途径以及EGb761的保护作用。
